The presentation has information regarding the components and proteins in milk which has biological importance. It also includes the methods to separate them from milk with the aid of the different membrane techniques.
The storage of human milk impacts its constituents depending on the storage method. Glass is the best container as it is least destructive. Milk can be safely refrigerated for 72 hours with little change to constituents. Freezing destroys cellular activity and reduces some vitamins. Boiling destroys some enzymes and reduces some immunological properties. Overall, the nutrient content is largely unchanged by storage but the immunological components are reduced depending on the storage technique used.
Detection of the Antibacterial Activity of Bioactive Peptide Isolated from Fe...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
This study examined the effects of high pressure processing (193 MPa at -20°C) versus low-temperature long-time (LTLT) pasteurization on various components in human breastmilk, including fatty acids, lipid oxidation products, total antioxidant capacity, and vitamin C content. It found that high pressure processing did not significantly change fatty acid content or composition, lipid oxidation levels, or total antioxidant capacity, though it did reduce ascorbic acid levels by over 11%. In contrast, LTLT pasteurization significantly reduced total vitamin C by 35% and ascorbic acid by 24%, with a minor 6% decrease in total antioxidant capacity. This suggests high pressure processing may better preserve some heat-labile
This study evaluated the pharmacokinetics and tissue irritation of two sulphadoxine-trimethoprim products (Products A and B) administered intravenously or subcutaneously in pre-ruminant calves. Product A caused more severe tissue irritation and higher creatine kinase levels at injection sites compared to Product B, though the difference was not statistically significant. Both products showed slower absorption and elimination of trimethoprim when given subcutaneously versus intravenously. The study suggests intravenous administration or a minimum subcutaneous dosage of 30 mg/kg every 24 hours or 15 mg/kg every 12 hours for pre-ruminant calves to effectively treat infections while avoiding marked tissue irritation.
Pharma supplements recombinant proteins as excipients Stephen Berezenko
1) The document discusses challenges in formulating biotherapeutics like proteins and how commonly used excipients like gelatin and human serum albumin have issues related to purity, consistency, and potential risk of transmitting diseases.
2) It describes how recombinant DNA technology has been used to produce recombinant versions of gelatin and human serum albumin as excipients that are highly pure, consistent, and avoid risks from animal or human sources.
3) Recombinant human albumin called Recombumin has been commercially developed and clinically tested as a safer alternative to use in biotherapeutic formulations compared to traditional excipients from animal or human sources.
Novozymes Veltis® – Engineerd albumins for optimized drug dosingNiklas Andersson
Novozymes Veltis® is a clinically-proven half-life extension technology based on engineered albumins that allows drug developers to optimize dose size and frequency to achieve improved patient compliance.
Biochemical and pharmacological study of biologically active preparation of p...inventionjournals
Our aim was to perform some biochemical and pharmacological studies of bioactive bovine placental preparation via digestion of cow placenta using enzyme contained in swine stomach. Amino acid compositions and contents in biologically active preparation of placenta, obtained by digestion of cow placenta with enzyme contained in swine stomach were measured by HPLC technique and it was found that contents of such amino acids as glycine, proline and lysine were highest and 9 essential amino acids, including valine, histidine, methionine, lysine, threonine, arginine, phenylalanine, leucine and isoleucine were measured. In pharmacological study, acute toxicity (LD50) of the preparation and effect of the preparation on immune response to sheep erythrocyte were investigated in white mice, weighing 18 to 20 g each. The study revealed acute toxicity (LD50) of the preparation was 60 ml per kg. Spleen index of the first and second experimental group animals treated by the preparation during both provoked and unprovoked immune responses increased by 1.2 to 3.09 times as compared to that of negative control animals, while splenocyte count elevated by 1.2 to 2.2 times than negative control animals. Higher contents of essential amino acids of the biologically active preparation of cattle placenta shows its biologically higher nutritive value, as well as pharmacological study reveals the preparation has minimal toxicity and higher effect to stimulate immune responses.
The document describes optimization of protein extraction from sacha inchi (Plukenetia volubilis) kernel cake using alkaline and enzyme-assisted methods. Response surface methodology was used to optimize extraction parameters. For alkaline extraction, optimal conditions were 54.2°C, solvent/meal ratio of 42/1, 1.65M NaCl concentration at pH 9.5 for 30 minutes, yielding 29.7% protein. For enzyme extraction, optimal conditions were 5.6% enzyme concentration, 40.4 minutes extraction at pH 9.0 and 50°C, solvent/meal ratio of 50/1, yielding 44.7% protein and 7.8% hydrolysis degree. The enzyme method produced higher protein recovery
The storage of human milk impacts its constituents depending on the storage method. Glass is the best container as it is least destructive. Milk can be safely refrigerated for 72 hours with little change to constituents. Freezing destroys cellular activity and reduces some vitamins. Boiling destroys some enzymes and reduces some immunological properties. Overall, the nutrient content is largely unchanged by storage but the immunological components are reduced depending on the storage technique used.
Detection of the Antibacterial Activity of Bioactive Peptide Isolated from Fe...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
This study examined the effects of high pressure processing (193 MPa at -20°C) versus low-temperature long-time (LTLT) pasteurization on various components in human breastmilk, including fatty acids, lipid oxidation products, total antioxidant capacity, and vitamin C content. It found that high pressure processing did not significantly change fatty acid content or composition, lipid oxidation levels, or total antioxidant capacity, though it did reduce ascorbic acid levels by over 11%. In contrast, LTLT pasteurization significantly reduced total vitamin C by 35% and ascorbic acid by 24%, with a minor 6% decrease in total antioxidant capacity. This suggests high pressure processing may better preserve some heat-labile
This study evaluated the pharmacokinetics and tissue irritation of two sulphadoxine-trimethoprim products (Products A and B) administered intravenously or subcutaneously in pre-ruminant calves. Product A caused more severe tissue irritation and higher creatine kinase levels at injection sites compared to Product B, though the difference was not statistically significant. Both products showed slower absorption and elimination of trimethoprim when given subcutaneously versus intravenously. The study suggests intravenous administration or a minimum subcutaneous dosage of 30 mg/kg every 24 hours or 15 mg/kg every 12 hours for pre-ruminant calves to effectively treat infections while avoiding marked tissue irritation.
Pharma supplements recombinant proteins as excipients Stephen Berezenko
1) The document discusses challenges in formulating biotherapeutics like proteins and how commonly used excipients like gelatin and human serum albumin have issues related to purity, consistency, and potential risk of transmitting diseases.
2) It describes how recombinant DNA technology has been used to produce recombinant versions of gelatin and human serum albumin as excipients that are highly pure, consistent, and avoid risks from animal or human sources.
3) Recombinant human albumin called Recombumin has been commercially developed and clinically tested as a safer alternative to use in biotherapeutic formulations compared to traditional excipients from animal or human sources.
Novozymes Veltis® – Engineerd albumins for optimized drug dosingNiklas Andersson
Novozymes Veltis® is a clinically-proven half-life extension technology based on engineered albumins that allows drug developers to optimize dose size and frequency to achieve improved patient compliance.
Biochemical and pharmacological study of biologically active preparation of p...inventionjournals
Our aim was to perform some biochemical and pharmacological studies of bioactive bovine placental preparation via digestion of cow placenta using enzyme contained in swine stomach. Amino acid compositions and contents in biologically active preparation of placenta, obtained by digestion of cow placenta with enzyme contained in swine stomach were measured by HPLC technique and it was found that contents of such amino acids as glycine, proline and lysine were highest and 9 essential amino acids, including valine, histidine, methionine, lysine, threonine, arginine, phenylalanine, leucine and isoleucine were measured. In pharmacological study, acute toxicity (LD50) of the preparation and effect of the preparation on immune response to sheep erythrocyte were investigated in white mice, weighing 18 to 20 g each. The study revealed acute toxicity (LD50) of the preparation was 60 ml per kg. Spleen index of the first and second experimental group animals treated by the preparation during both provoked and unprovoked immune responses increased by 1.2 to 3.09 times as compared to that of negative control animals, while splenocyte count elevated by 1.2 to 2.2 times than negative control animals. Higher contents of essential amino acids of the biologically active preparation of cattle placenta shows its biologically higher nutritive value, as well as pharmacological study reveals the preparation has minimal toxicity and higher effect to stimulate immune responses.
The document describes optimization of protein extraction from sacha inchi (Plukenetia volubilis) kernel cake using alkaline and enzyme-assisted methods. Response surface methodology was used to optimize extraction parameters. For alkaline extraction, optimal conditions were 54.2°C, solvent/meal ratio of 42/1, 1.65M NaCl concentration at pH 9.5 for 30 minutes, yielding 29.7% protein. For enzyme extraction, optimal conditions were 5.6% enzyme concentration, 40.4 minutes extraction at pH 9.0 and 50°C, solvent/meal ratio of 50/1, yielding 44.7% protein and 7.8% hydrolysis degree. The enzyme method produced higher protein recovery
This document presents a simplified and efficient process for producing insulin in Pichia pastoris yeast. Key points:
- Insulin production is currently done in E. coli or Saccharomyces cerevisiae, but the S. cerevisiae process has up to 15 purification steps.
- The present study uses P. pastoris, which secretes correctly folded insulin directly into culture medium. A high-density fermentation achieved a high yield of 2.26 g/L of insulin precursor.
- A novel two-step purification process using tangential flow filtration and cation exchange chromatography achieved high loading capacity and purity.
- The purified insulin precursor was then converted to human insulin through an enzymatic
The Electrophoretic Profile Myofibrillar Proteins Extracted From Camel Muscle...IJEAB
Changes in electrophoretic profiles of myofibrillar protein (MFP) in the Longissimus thoracis (LD) of young camels (2 to 4 years), preserved by refrigeration has been treated or not by lactic acid solution 4% or citric acid 1%, were followed during the post-mortem time at the following times: 1, 2, 4, 6, 8, 10, 12, 24 and 48 hours. The cold preservation for 48 hours has not shown any particular distinctions in the protein profiles of this muscle. Changes related to the type of treatment were recorded during the storage time. Proteolysis of the myofibrillar fraction was earlier in this muscle in the case of treatment with one of two solutions of organic acids used, particularly in the case of using lactic acid. Indeed, these changes have affected at the first hour after slaughter the proteolysis of the myofibrillar proteins. Fragments of low molecular weight (42, 36, 33, 26, 23, 18, 16, 14 and 13 kDa) have been identified. The electrophoretic analysis showed that during refrigeration, LD treated with a solution of lactic acid is more sensitive to disruption phenomena and muscle protein proteolysis that lots of this muscle that even in the case of preservation by refrigeration only or by refrigeration
The document summarizes a presentation on antioxidant peptides derived from the blue-spotted stingray. It discusses alternative methods to obtain bioactive peptides from proteins, characterization of antioxidant capacity, identification of two antioxidant peptides (WAFAPA and MYPGLA) from the <3 kDa fraction of stingray hydrolysate, and characterization of their stability under thermal, pH, and simulated digestion conditions. The conclusion discusses the potential of antioxidant peptides for food and medical applications and need for further research.
Microbial enzymes are widely used in food processing due to their ability to catalyze specific reactions. About 80% of total enzyme production by dollar value is used by the food industry. Enzymes offer advantages over whole microorganisms by catalyzing single step conversions of specific substrates to products. The main classes of enzymes used in food processing are hydrolases, isomerases, and oxidoreductases. Recombinant DNA technology allows for improved production of enzymes in bacteria and yeast. Immobilizing enzymes allows for their reuse, reducing costs. Thermostable enzymes are advantageous as they maintain activity at higher temperatures, increasing reaction rates. Enzymes are also used to treat food waste by converting components into value-added
Recombinant dna technology for food usesUzair Hashmi
Recombinant DNA technology allows for the production of novel enzymes suited for specific food processing conditions. This is done by screening microorganisms from diverse environments and modifying known enzymes. Recombinant production offers advantages like increased enzyme yields and removal of toxic metabolites. An example is improving pectinase production in Penicillium griseoroseum through recombinant techniques, increasing polygalacturonase and pectin lyase yields by 143-fold and 15-fold respectively. Chymosin, used in cheese production, has also been recombinantly produced in Kluyveromyces lactis as an alternative to animal-sourced rennet. A comparative study found recombinant goat chymosin performed best with the highest activity
MaxiChick™ is the patented combination of two scientifically proven DSM products (Hy•D® and Carophyll® Red) that complement each other to boost the productivity of the breeder flock. An efficient source of Vitamin D3 and carotenoids for pigmentation are the two compounds of MaxiChick.
To find out more about DSM's work on poultry visit us on:http://www.dsm.com/markets/anh/en_US/species/species-poultry.html
Or why not speak to a specialist on Twitter: @goldenyolk
This study investigated the prevalence of Pseudomonas species in raw cow and buffalo milk samples from Assiut, Egypt. Pseudomonas was found in 40% of cow milk samples and 50% of buffalo milk samples. Five Pseudomonas species were identified: P. aeruginosa, P. fluorescens, P. putida, P. cepacia, and P. stutzeri. The study also examined the effect of storage temperature (4°C and -18°C) on the growth and protease/lipase enzyme activity of Pseudomonas isolates over 14 days. Pseudomonas grew continuously at 4°C for 7 days then was destroyed, while at -18°C growth and enzyme
Hydrolysate Characterization Technical Presentation Webinar11 2009Mason Williams
1) The document discusses developing a chemically defined hydrolysate alternative to mitigate risks associated with undefined natural hydrolysates.
2) The approach involves fractionating multiple hydrolysates using RP-HPLC and identifying bioactive fractions that stimulate cell growth and protein production in CHO cells.
3) A new product called EX-CELL CD Hydrolysate Fusion was developed that contains synthetic components identified from bioactive fractions and provides 80-100% of the performance of natural hydrolysates.
This document discusses enzymatic tailoring of hydrolysate properties. It provides an overview of enzymes, including enzyme classes and nomenclature. It describes how enzymatic hydrolysis can be used to customize the chemical and biological properties of protein hydrolysates, such as solubility, molecular weight distribution, and amino acid profile. Several examples are given of how different proteases can be used to tailor the degree of hydrolysis, antioxidant activity, foam stability, and amino acid composition of soy flour hydrolysates. The document concludes that enzymatic tailoring provides opportunities to engineer product specifications, improve yields and recoveries, customize protein properties, and develop new active ingredients and product paradigms from plant sources.
Presented at a DSM customer event organized in Village Neuf, France on June 2013: Exploring the benefits of feed carotenoids for egg quality.
Yves Nys-Feeding laying hens and egg quality-Focus on carotenoids.
ND Pharma & Biotech develops a product called PreserFood TM ®, which is added to milk to preserve it for up to 42 days by inhibiting bacterial growth. The compound is made from a nonessential amino acid and hydroxytyrosol from olives. It improves milk flavor and extends shelf life without needing preservatives. PreserFood TM ® allows milk to stay fresh longer while maintaining nutritional quality.
Alpha Lactalbumin - Development path for a valuable dairy protein ingredientJuan Gonzalez
Provides a multifunctional view of the drivers behind the development and application of alpha lactalbumin as a valuable dairy protein ingredient in nutritional foods
PPT: Supplementing an Immunomodulatory Feed Ingredient to Modulate thermoregu...Faisal A. Alshamiry
Supplementing an Immunomodulatory Feed Ingredient to Modulate Thermoregulation, Physiologic, & Production Responses in Lactating Dairy Cows Under Heat Stress Conditions
1) The study investigated the potential protective effects of honey against paracetamol-induced liver toxicity in rats.
2) Administration of paracetamol caused significant increases in liver enzymes and oxidative stress markers along with liver damage.
3) Pretreatment with honey or the reference drug silymarin prevented the paracetamol-induced increases in liver enzymes and oxidative stress and reduced liver damage based on histological examination.
Food preservation by high pressure - Heinz 2009Wouter de Heij
This document summarizes key aspects of using high pressure processing (HPP) to preserve foods. HPP uses hydrostatic pressure above 350 MPa to inactivate microorganisms and enzymes at low temperatures, preserving nutrients, flavors, and textures. HPP is considered an alternative to thermal pasteurization. It allows for short processing times under 5 minutes with low thermal impact and energy use. HPP selectively inactivates bacteria, spores, and viruses through physical and chemical changes induced by high pressure. The effectiveness of HPP results from compressing foods by up to 25% of their volume. International use of HPP in food processing has been growing for preservation of foods like meats, seafood, fruits, and vegetables
Preservation of Meat Using High Pressure Processingnavi04vet
This document discusses the use of high pressure processing (HPP) to preserve meat. It begins with an introduction to food preservation methods and a brief history of HPP. The working principles of HPP are then explained. The document outlines various effects of HPP on physico-chemical properties of meat including tenderization, water retention, microbial inactivation, and oxidation. It provides examples of HPP applications for different meat products and major HPP industries worldwide. In conclusion, HPP is presented as a promising preservation method that extends shelf life without compromising nutrition, though it can impact color and enhance oxidation requiring added antioxidants.
IMMUNOCAL is a patented bovine whey protein isolate that provides cysteine, a precursor for intracellular glutathione synthesis. Glutathione plays a key role in the immune system and cellular antioxidant defenses. IMMUNOCAL increases glutathione levels by supplying cysteine, which is typically limited in the diet. Clinical studies have shown IMMUNOCAL to be safe and well-tolerated for supplementing glutathione in conditions involving oxidative stress, immune deficiency, and xenobiotic overload.
Microbiological and physicochemical quality of pasteurized milk supplemented ...UniversitasGadjahMada
Caesalpinia sappan L (Sappanwood) contains antibacterial compounds and antioxidants that inhibit the growth of microbes. This study aimed to investigatethe microbiological and physicochemical qualities of pasteurized milk supplemented with 0, 2, 4, 6 and 8% (w/v) sappan wood extract. Data were analyzed using a completely randomized design factorial followed by the Duncan’s new multiple range test. Preliminary analysis showed that sappan wood extract contained 44.66 ± 0.09 mg/100g phenols, 0.18 ± 0.01 mg/100mg flavonoids, 46.42 ± 0.23 mg/100g tannins, and antioxidant activity at 85.82 ± 0.25%. The addition of sappan wood extract significantly increased the antioxidant activity (P<0.05) of pasteurized milk during storage. Pasteurized milk supplemented with sappan wood extract had a lower total bacterial count (P<0.05) than that of unsupplemented pasteurized milk, and supplemented milk showed strong antibacterial activities against Escherichia coli, Shigella flexneri, Salmonella thypimurium, Staphylococcus aureus, and Listeria monocytogenes.The addition of sappan wood slightly increased the protein content but did not affect pH, and viscosity. It is concluded that the addition of sappan wood extract increased the microbiological quality and maintained the physicochemical quality of pasteurized milk, thus extending the product’s shelf-life.
Application of ultrafiltration technique for the quality improvement of dahi ...Ganga Sahay Meena
This document describes a study on using ultrafiltration techniques to improve the quality of dahi, a fermented milk product from India. Ultrafiltered buffalo milk with varying protein levels was used to produce dahi. Dahi made from ultrafiltered milk showed increased firmness, stickiness, sensory scores and decreased whey separation compared to the control dahi made without ultrafiltration. Principal component analysis revealed that protein content was positively correlated with attributes like firmness while negatively correlated with whey separation. Overall, ultrafiltration improved the quality attributes of dahi by increasing the protein levels in milk.
Impact of non-processing technology in dairy products for microbial safety | ...FoodresearchLab
Dairy products, especially milk is highly perishable as it contains ample nutrition and high in moisture content for the microorganism to grow and multiply.
1.Pulsed electric Field (PEF)
2.High Pressure Processing (HPP)
3.Ultrasound (US)
4.Plasma and low plasma Technology (PT)
To Read More : https://bit.ly/2UX13af
Most commercial fermented milks showed moderate ACE-inhibitory activity, though some exceptions had higher activity that could relate to the milk origin. Two fermented milks were subjected to simulated gastrointestinal digestion, which increased their ACE-inhibitory activity, suggesting physiological digestion promotes formation of active peptides. Peptides generated from one product during digestion were sequenced, and most formed after 30 minutes of pancreatic enzyme incubation, indicating digestion facilitates peptide release from proteins in these fermented products.
This document presents a simplified and efficient process for producing insulin in Pichia pastoris yeast. Key points:
- Insulin production is currently done in E. coli or Saccharomyces cerevisiae, but the S. cerevisiae process has up to 15 purification steps.
- The present study uses P. pastoris, which secretes correctly folded insulin directly into culture medium. A high-density fermentation achieved a high yield of 2.26 g/L of insulin precursor.
- A novel two-step purification process using tangential flow filtration and cation exchange chromatography achieved high loading capacity and purity.
- The purified insulin precursor was then converted to human insulin through an enzymatic
The Electrophoretic Profile Myofibrillar Proteins Extracted From Camel Muscle...IJEAB
Changes in electrophoretic profiles of myofibrillar protein (MFP) in the Longissimus thoracis (LD) of young camels (2 to 4 years), preserved by refrigeration has been treated or not by lactic acid solution 4% or citric acid 1%, were followed during the post-mortem time at the following times: 1, 2, 4, 6, 8, 10, 12, 24 and 48 hours. The cold preservation for 48 hours has not shown any particular distinctions in the protein profiles of this muscle. Changes related to the type of treatment were recorded during the storage time. Proteolysis of the myofibrillar fraction was earlier in this muscle in the case of treatment with one of two solutions of organic acids used, particularly in the case of using lactic acid. Indeed, these changes have affected at the first hour after slaughter the proteolysis of the myofibrillar proteins. Fragments of low molecular weight (42, 36, 33, 26, 23, 18, 16, 14 and 13 kDa) have been identified. The electrophoretic analysis showed that during refrigeration, LD treated with a solution of lactic acid is more sensitive to disruption phenomena and muscle protein proteolysis that lots of this muscle that even in the case of preservation by refrigeration only or by refrigeration
The document summarizes a presentation on antioxidant peptides derived from the blue-spotted stingray. It discusses alternative methods to obtain bioactive peptides from proteins, characterization of antioxidant capacity, identification of two antioxidant peptides (WAFAPA and MYPGLA) from the <3 kDa fraction of stingray hydrolysate, and characterization of their stability under thermal, pH, and simulated digestion conditions. The conclusion discusses the potential of antioxidant peptides for food and medical applications and need for further research.
Microbial enzymes are widely used in food processing due to their ability to catalyze specific reactions. About 80% of total enzyme production by dollar value is used by the food industry. Enzymes offer advantages over whole microorganisms by catalyzing single step conversions of specific substrates to products. The main classes of enzymes used in food processing are hydrolases, isomerases, and oxidoreductases. Recombinant DNA technology allows for improved production of enzymes in bacteria and yeast. Immobilizing enzymes allows for their reuse, reducing costs. Thermostable enzymes are advantageous as they maintain activity at higher temperatures, increasing reaction rates. Enzymes are also used to treat food waste by converting components into value-added
Recombinant dna technology for food usesUzair Hashmi
Recombinant DNA technology allows for the production of novel enzymes suited for specific food processing conditions. This is done by screening microorganisms from diverse environments and modifying known enzymes. Recombinant production offers advantages like increased enzyme yields and removal of toxic metabolites. An example is improving pectinase production in Penicillium griseoroseum through recombinant techniques, increasing polygalacturonase and pectin lyase yields by 143-fold and 15-fold respectively. Chymosin, used in cheese production, has also been recombinantly produced in Kluyveromyces lactis as an alternative to animal-sourced rennet. A comparative study found recombinant goat chymosin performed best with the highest activity
MaxiChick™ is the patented combination of two scientifically proven DSM products (Hy•D® and Carophyll® Red) that complement each other to boost the productivity of the breeder flock. An efficient source of Vitamin D3 and carotenoids for pigmentation are the two compounds of MaxiChick.
To find out more about DSM's work on poultry visit us on:http://www.dsm.com/markets/anh/en_US/species/species-poultry.html
Or why not speak to a specialist on Twitter: @goldenyolk
This study investigated the prevalence of Pseudomonas species in raw cow and buffalo milk samples from Assiut, Egypt. Pseudomonas was found in 40% of cow milk samples and 50% of buffalo milk samples. Five Pseudomonas species were identified: P. aeruginosa, P. fluorescens, P. putida, P. cepacia, and P. stutzeri. The study also examined the effect of storage temperature (4°C and -18°C) on the growth and protease/lipase enzyme activity of Pseudomonas isolates over 14 days. Pseudomonas grew continuously at 4°C for 7 days then was destroyed, while at -18°C growth and enzyme
Hydrolysate Characterization Technical Presentation Webinar11 2009Mason Williams
1) The document discusses developing a chemically defined hydrolysate alternative to mitigate risks associated with undefined natural hydrolysates.
2) The approach involves fractionating multiple hydrolysates using RP-HPLC and identifying bioactive fractions that stimulate cell growth and protein production in CHO cells.
3) A new product called EX-CELL CD Hydrolysate Fusion was developed that contains synthetic components identified from bioactive fractions and provides 80-100% of the performance of natural hydrolysates.
This document discusses enzymatic tailoring of hydrolysate properties. It provides an overview of enzymes, including enzyme classes and nomenclature. It describes how enzymatic hydrolysis can be used to customize the chemical and biological properties of protein hydrolysates, such as solubility, molecular weight distribution, and amino acid profile. Several examples are given of how different proteases can be used to tailor the degree of hydrolysis, antioxidant activity, foam stability, and amino acid composition of soy flour hydrolysates. The document concludes that enzymatic tailoring provides opportunities to engineer product specifications, improve yields and recoveries, customize protein properties, and develop new active ingredients and product paradigms from plant sources.
Presented at a DSM customer event organized in Village Neuf, France on June 2013: Exploring the benefits of feed carotenoids for egg quality.
Yves Nys-Feeding laying hens and egg quality-Focus on carotenoids.
ND Pharma & Biotech develops a product called PreserFood TM ®, which is added to milk to preserve it for up to 42 days by inhibiting bacterial growth. The compound is made from a nonessential amino acid and hydroxytyrosol from olives. It improves milk flavor and extends shelf life without needing preservatives. PreserFood TM ® allows milk to stay fresh longer while maintaining nutritional quality.
Alpha Lactalbumin - Development path for a valuable dairy protein ingredientJuan Gonzalez
Provides a multifunctional view of the drivers behind the development and application of alpha lactalbumin as a valuable dairy protein ingredient in nutritional foods
PPT: Supplementing an Immunomodulatory Feed Ingredient to Modulate thermoregu...Faisal A. Alshamiry
Supplementing an Immunomodulatory Feed Ingredient to Modulate Thermoregulation, Physiologic, & Production Responses in Lactating Dairy Cows Under Heat Stress Conditions
1) The study investigated the potential protective effects of honey against paracetamol-induced liver toxicity in rats.
2) Administration of paracetamol caused significant increases in liver enzymes and oxidative stress markers along with liver damage.
3) Pretreatment with honey or the reference drug silymarin prevented the paracetamol-induced increases in liver enzymes and oxidative stress and reduced liver damage based on histological examination.
Food preservation by high pressure - Heinz 2009Wouter de Heij
This document summarizes key aspects of using high pressure processing (HPP) to preserve foods. HPP uses hydrostatic pressure above 350 MPa to inactivate microorganisms and enzymes at low temperatures, preserving nutrients, flavors, and textures. HPP is considered an alternative to thermal pasteurization. It allows for short processing times under 5 minutes with low thermal impact and energy use. HPP selectively inactivates bacteria, spores, and viruses through physical and chemical changes induced by high pressure. The effectiveness of HPP results from compressing foods by up to 25% of their volume. International use of HPP in food processing has been growing for preservation of foods like meats, seafood, fruits, and vegetables
Preservation of Meat Using High Pressure Processingnavi04vet
This document discusses the use of high pressure processing (HPP) to preserve meat. It begins with an introduction to food preservation methods and a brief history of HPP. The working principles of HPP are then explained. The document outlines various effects of HPP on physico-chemical properties of meat including tenderization, water retention, microbial inactivation, and oxidation. It provides examples of HPP applications for different meat products and major HPP industries worldwide. In conclusion, HPP is presented as a promising preservation method that extends shelf life without compromising nutrition, though it can impact color and enhance oxidation requiring added antioxidants.
IMMUNOCAL is a patented bovine whey protein isolate that provides cysteine, a precursor for intracellular glutathione synthesis. Glutathione plays a key role in the immune system and cellular antioxidant defenses. IMMUNOCAL increases glutathione levels by supplying cysteine, which is typically limited in the diet. Clinical studies have shown IMMUNOCAL to be safe and well-tolerated for supplementing glutathione in conditions involving oxidative stress, immune deficiency, and xenobiotic overload.
Microbiological and physicochemical quality of pasteurized milk supplemented ...UniversitasGadjahMada
Caesalpinia sappan L (Sappanwood) contains antibacterial compounds and antioxidants that inhibit the growth of microbes. This study aimed to investigatethe microbiological and physicochemical qualities of pasteurized milk supplemented with 0, 2, 4, 6 and 8% (w/v) sappan wood extract. Data were analyzed using a completely randomized design factorial followed by the Duncan’s new multiple range test. Preliminary analysis showed that sappan wood extract contained 44.66 ± 0.09 mg/100g phenols, 0.18 ± 0.01 mg/100mg flavonoids, 46.42 ± 0.23 mg/100g tannins, and antioxidant activity at 85.82 ± 0.25%. The addition of sappan wood extract significantly increased the antioxidant activity (P<0.05) of pasteurized milk during storage. Pasteurized milk supplemented with sappan wood extract had a lower total bacterial count (P<0.05) than that of unsupplemented pasteurized milk, and supplemented milk showed strong antibacterial activities against Escherichia coli, Shigella flexneri, Salmonella thypimurium, Staphylococcus aureus, and Listeria monocytogenes.The addition of sappan wood slightly increased the protein content but did not affect pH, and viscosity. It is concluded that the addition of sappan wood extract increased the microbiological quality and maintained the physicochemical quality of pasteurized milk, thus extending the product’s shelf-life.
Application of ultrafiltration technique for the quality improvement of dahi ...Ganga Sahay Meena
This document describes a study on using ultrafiltration techniques to improve the quality of dahi, a fermented milk product from India. Ultrafiltered buffalo milk with varying protein levels was used to produce dahi. Dahi made from ultrafiltered milk showed increased firmness, stickiness, sensory scores and decreased whey separation compared to the control dahi made without ultrafiltration. Principal component analysis revealed that protein content was positively correlated with attributes like firmness while negatively correlated with whey separation. Overall, ultrafiltration improved the quality attributes of dahi by increasing the protein levels in milk.
Impact of non-processing technology in dairy products for microbial safety | ...FoodresearchLab
Dairy products, especially milk is highly perishable as it contains ample nutrition and high in moisture content for the microorganism to grow and multiply.
1.Pulsed electric Field (PEF)
2.High Pressure Processing (HPP)
3.Ultrasound (US)
4.Plasma and low plasma Technology (PT)
To Read More : https://bit.ly/2UX13af
Most commercial fermented milks showed moderate ACE-inhibitory activity, though some exceptions had higher activity that could relate to the milk origin. Two fermented milks were subjected to simulated gastrointestinal digestion, which increased their ACE-inhibitory activity, suggesting physiological digestion promotes formation of active peptides. Peptides generated from one product during digestion were sequenced, and most formed after 30 minutes of pancreatic enzyme incubation, indicating digestion facilitates peptide release from proteins in these fermented products.
The application of high pressure processing in dairy industry (Presentation)ESTIA LABORATORY
This document discusses the application of high pressure processing (HPP) in the dairy industry. HPP is a non-thermal pasteurization method that kills microbes while retaining the organoleptic and nutritional attributes of food. It has several applications in dairy including milk homogenization, emulsion production like chocolate milk, cheese production, and processing of dairy wastewater to recover proteins and lactose. HPP is presented as an alternative to thermal processing for applications like pasteurization and sterilization that cause nutritional and quality losses. The document provides examples of HPP use in emulsification, wastewater fractionation, and cheese production through ultrafiltration. It concludes that HPP will allow for the production of safer, healthier
The application of high pressure processing in dairy industry (Presentation)Thomas Zafeiriadis
This document discusses the application of high pressure processing (HPP) in the dairy industry. It begins with an introduction to HPP as a non-thermal pasteurization method that retains nutrients and organoleptic properties unlike conventional heating. It then discusses several applications of HPP including milk homogenization, emulsion production like chocolate milk, cheese production, and managing dairy wastewater to recover proteins and lactose.
NIZO Plant Protein Functionality Conference on October 21-22 gathered around 450 attendees to discuss the recent findings and innovations on plant proteins. Research team leader Emilia Nordlund gave a keynote presentation on bioprocessing technologies to improve the plant protein functionality.
PROCESS OF EXTRACTION OF LACTOPEROXIDASE PROTEIN FROM YOGURT WHEYAin Nur Syazwani
This document provides information about extracting lactoperoxidase protein from yogurt whey. It begins with an introduction to lactoperoxidase and its properties. It then discusses the market survey and potential applications. A process flow diagram is presented, showing the upstream and downstream processing steps. Key steps include centrifugation, homogenization, fermentation, microfiltration, ion exchange chromatography, and spray drying. Section 1 discusses pre-treatment including milk standardization. Section 2 covers downstream processing, comparing methods for separating insolubles like ion exchange, microfiltration, and ultrafiltration, selecting microfiltration for this project.
The Lactoperoxidase system (LP-s) consists of the production of an antibacterial compound from the thiocyanate ion catalysed by Lactoperoxidase in the presence of hydrogen peroxide.
This document summarizes the process of producing human insulin through recombinant DNA technology using E. coli bacteria. It involves growing E. coli in a bioreactor to produce proinsulin inclusion bodies, isolating the inclusion bodies through centrifugation, solubilizing and refolding the proinsulin, and purifying it through affinity chromatography, site-specific cleavage, reverse phase chromatography, and polishing. The purified human insulin can then be stored at 4°C for pharmaceutical use in diabetes treatment.
Milk contains many proteins that have health benefits. Common techniques used in the dairy industry to extract milk proteins include precipitation, membrane filtration, and chromatography. Precipitation involves adjusting factors like pH, salts, or temperature to make proteins insoluble and aggregate. Membrane filtration separates components based on size or hydrophobicity. Chromatography techniques like ion exchange and affinity chromatography isolate proteins based on their charge or affinity for specific chemical groups.
Recent Advances in Dairy Industry -Chirag Prajapati.pptxChirag Prajapati
Embark on a journey through the realm of "Recent Advances in Dairy Processing" with this enlightening PowerPoint presentation. Discover the cutting-edge technologies and innovations transforming the dairy industry. Learn about novel processing methods, advanced equipment, and sustainable practices that are revolutionizing milk and dairy product manufacturing. From ultra-high-temperature processing (UHT) to membrane filtration and beyond, this presentation highlights the latest developments that enhance efficiency, quality, and sustainability in dairy processing.
Microbial fermentation By Aneela SaleemAneelaSaleem
This document discusses different types of fermentation processes used in industry. It begins with an introduction and overview of fermentation media and microorganisms. It then describes the main types of fermentation processes - batch, fed-batch, and continuous fermentation - and factors that influence each type such as growth rate and flow rate. The document also covers solid state and submerged liquid fermentations. Important considerations for continuous fermentation are highlighted. Recent advances in fermentation technology are briefly mentioned at the end.
Advancement in research for improving the ambient temperature stability of a ...Arup Nag
An overview of my research in a Centre of Research Excellence in New Zealand to develop novel technology which successfuly enabled probiotic strains remain stable for longer duration under ambient storage conditions.
This document summarizes a study that aimed to develop an extract of bambara nut (Vigna subterranea) protein for use as a reagent in skin prick tests (SPTs) to detect food allergies. Bambara nut protein was extracted via isoelectric precipitation and characterized via SDS-PAGE, identifying 14 bands between 17-122 kDa. The extract was formulated into an SPT reagent and tested on 11 individuals with food allergies and 9 without, measuring wheal responses. Total and bambara nut-specific IgE in subject sera were also analyzed. Two allergic subject sera showed specific IgE binding to bambara nut allergen by immunoblotting
ABSTRACT Human platelet-derived growth factor-BB (hPDGF-BB), a proliferation factor, has been successfully manufactured and approved by FDA as the treatment for diabetic foot ulcer and self bone grafting. There have been no reports on the storage of the recombinant hPDGF-BB (rhPDGF-BB) in the Pichia pastoris fermentation broth although during the research of process development and the production manufacture it needs to be stored at low temperature. The concentration of rhPDGF-BB protein in the fed-batch fermentation broth of P. pastoris was stable during 3-week storage at -20oC, but its bioactivity was reduced by 20%. The addition of a mixture of 50% glycerol with either 1 mM EDTA or 1 mM PMSF into the fermentation broth could fully preserve the bioactivity of rhPDGF-BB until 3 weeks at -20oC. The addition of 50% glycerol with either 1 mM EDTA or 1 mM PMSF was found no affection on the protein purification process.
Key-words: rhPDGF-BB, Pichia pastoris, Fed-batch fermentation, Protein stability, Bioactivity, Storage
Uf permeate is rich source of essential nutrients like whey protein, minerals, vitamins etc., therefore it provides great scope in the manufacturing of sports drinks, alcoholic bevarages and so on...
The presentation will focus on the application of the FoodPro® Cleanline enzyme Glycerophospholipid Cholesterol Acyltransferase (GCAT) in the milk, which converts cholesterol to cholesterol esters and lysophospholipids. Lysophospholipids are surface-active amphiphiles that will act as biosurfactants and prevent protein aggregation and promote refolding of proteins (chaperone-like properties), resulting in various processing advantages and increased emulsion stability when used in the manufacture of milk and dairy products. GCAT can reduce free cholesterol in milk by as much as 65 to 85%, which could potentially be an important advantage from a nutritional point of view when producing milk and dairy products.
ANTIOXIDANT ACTIVITY AND HEPATOPROTECTIVE EFFECTOF POMEGRANATE PEEL AND WHEY...Anurag Raghuvanshi
The antioxidant activity of pomegranate peel powder (PPP) and whey powder (WP) was evaluated, their hepatoprotective effect of each alone or in combination (PPWP) at equal levels was also evaluated in Wistar rats against carbon tetrachloride (CCL4) induced liver injury.
The hepatoprotective activity was assessed using various biochemical parameters and histopathological studies.
Novel functional fermented dairy product rich in menaquinone-7Rachel Southee
This document describes research into developing a novel functional dairy product rich in menaquinone-7 (MK-7) through fermentation using Bacillus subtilis natto. The maximum MK-7 concentration of 11.22 mg/L was achieved using standard milk with 1% glycerol at 40°C, 200 rpm, and 1 vvm air flow for 48 hours of fermentation. MK-7 production increased during the cell's exponential growth phase and peaked when the glycerol carbon source was exhausted after 30 hours. This research demonstrates the potential of using fermented milk to produce a product high in MK-7 and calcium to address bone and cardiovascular health issues.
Membrane technology in prophylactic biologicals in milk
1. Application of Membrane Technology in Prophylactic Biologicals in
Milk
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Presented by:
Parth Hirpara
Dairy Technology Dept.
2. What is Prophylactic Biologicals?
Prophylactic is defined as a preventive measure.
The word comes from the Greek for "an advance guard," an
suitable term for a measure taken to fend off a disease or
another unwanted consequence.
A prophylactic is a medication or a treatment designed and
used to prevent a disease from occurring.
(www.medicinenet.com)
Prophylactic biologicals can be defined as “ to prevent a
range of human ailments such as arthritis, toothache,
allergies, various kinds of viral infections such as common
cold, etc.”
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3. Why prophylactic biologicals from milk &
Membrane Technology?
Extra-nutritional role of milk proteins in maintaining the
physiological normalcy of the body at organ & cellular level
function.
Due to side effects of synthetic drugs as milk is a natural
source.
The physiological functions of the drug principle from milk
can be:
Get destroyed to various extent during routine processing (Depends
upon severity of the heat treatment).
Also influenced by environmental contaminants such as
organopesticides & insecticides.
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4. How to extract this biologicals?
Membrane processing of milk allows separation of insoluble
(fat & casein) fraction from soluble (whey proteins, Lactose,
peptides & NPN) fraction.
Casein & fat fraction processed separately, avoiding inevitable
interaction between the casein and whey protein fractions
under the influence of heat.
This allows reconstitution of milk with “Bio-protective factor”
intact.
Permeate whey proteins are further UF to get highly pure
undenatured whey protein isolates, which displays
prophylactic quality.
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5. 10/30/2018Membrane Technology in Dairy Processing
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Categories of
prophylactic
biologicals
GMP
Lf & LP
Bioactive
peptides
Phospholipids
6. Milk peptides with cardiovascular activity
There is a sequence homology between γ-fibrinogen in blood & κ-casein (GMP).
GMP:
Inhibits platelet aggregation.
Combine with receptor site & consequently prevents fibrinogen binding with blood
platelets (anti-thrombotic activity).
In vitro, anti-aggrigative properties is reinforced by the presence of Lys residue
in the sequence.
The 112-116 peptide resulting from the tryptic hydrolysis of GMP seems to be 200
fold more active than 113-116 sequence.
Hydrolysis of angiotensin-I by ACE is the key step in the physiological regulation
of blood pressure.
43-52 peptide of β-casein possess such activity.
All such peptides have mol. Wt. between 30-80kDa & primarily UF is used and
further they are isolated using SDS-PAGE.
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7. Biologically functional peptides derived from
milk proteins
1. Opioid agonist peptides bind to opioid receptors and
exhibits morphine like activity.
First characterized opioid agonist peptides from milk proteins
are:
β-casomorphin 7: Try-Pro-Phe-Pro-Gly-Pro-Ile (60-66th)
β-casomorphin 5: Try-Pro-Phe-Pro-Gly (60-64th)
β-casomorphin 5 is five times more active than BCM 7..
Similar peptides are present in αs-casein (90-96th) known as αs-
casein exorphin.
They are produced by chemical synthesis (isoelectric
precipitation) or enzymatic digestion.
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8. 2. Enzymatic hydrolysis of casein seems to produce ACE-
inhibitory peptides.
ACE converts angiotensin I to II; the later is very hypertensive peptide.
This enzyme also hydrolysis bradykinin, which is a hypotensive peptide.
Therefore ACE inhibitors are antihypertensive peptides.
ACEI from β-casein & κ-casein are most potent inhibitor is a
decapeptide (43-52).
3. Antimicrobial peptides:
Some fragments of β-casein gives phagocytosis stimulating peptides.
This peptides in vitro stimulates phagocytosis activity of murine &
human macrophages & exert in vivo a protective effect against
Klebsiella Pneumonia infection of mice.
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9. Bioactive components from cheese whey
Lf & LP have very high values as fine chemicals as well as functional foods
as well as pharmacological products.
Both Lf & LP have pI between 4.2-5.3.
For isolation of Lf & LP the whey is exposed to certain exchanger for
selective adsorption.
Use is made here of the net positive charge of the both Lf & LP in contrast
to the other whey proteins, which have net negative charges in this pH
range.
By charge interaction Lf & LP molecules binds to the negatively loaded
functional groups of the cation exchanger which leads to fixation of these
molecules on the ion exchange resins while the other whey proteins pass
through it (because they are negatively charged).
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10. Basic criterion for isolation of Lf & LP
Need of particle free whey (to keep high flow rate during the
loading phase) because very high volumes of whey have to
pass the ion exchange column.
LP in sweet whey is ~20mg/L & Lf is ~35mg/L.
So, theoretically for 100% yield: 50m of whey has to be
treated to get 1Kg of LP and 30m for 1Kg Lf.
The capacity of used ion exchange resin is equivalent to
adsorption of 40-45g of LP plus Lf from whey per litre of resin
can pass the ion exchange column during 15-20hr per ion
exchange cycle.
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11. Pharmaceutical application of Lf
Bio-protective nature of Lf.
Lf in infant formulas, health foods, skin creams as
antimicrobial product.
To treat leukemia (Cancer).
In association with the human γ-interferon, Lf supresses easily
human monocytes or mono-blastoid cell lines.
Lf also enhance the action of certain antibiotics which
enhances the efficacy of pharmaceutical preparations.
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13. Phospholipid from MFGM
Phospholipids and glycoproteins from the MFGM affect several
cell functions, such as growth, molecular transport system,
memory processing, stress responses, and central nervous
system myelination.
Have an important role as emulsifiers in food systems and can
be used to improve the features of bread, chocolate,
margarine and dairy products.
Use in a delivery or carrier system for drugs and other
substances.
By using membrane technology, 95% pure phospholipid can
be obtained.
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14. Use of ultrafiltration and supercritical fluid
extraction to obtain a whey buttermilk
powder enriched in milk fat globule membrane
phospholipids
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Marcelade Rezende Costa, Xiomara Elizabeth Elias-Argote, Rafael Jiménez-Flores and
Mirna Lúcia Gigante (2010)
ARTICLE1
15. Introduction
Whey buttermilk, a by-product from whey cream processing to butter, is rich
in milk fat globule membrane (MFGM) constituents, which have technological
and potential health properties.
Whey butter milk was concentrated by ultrafiltration (10X) and subsequently
Diafiltered (5X) (10kDa molecular mass cut off membrane) at 25oC and the
final retentate was spray dried.
The whey butter milk powder was submitted to supercritical
extraction(350bar,50oC) using carbon dioxide.
The membrane filtration removed most of the lactose and ash from the whey
buttermilk, and the super critical extraction extracted exclusively non-polar
lipids.
The final powder contained 73% Protein and 21% lipids, of which 61% were
phospholipids.
This ingredient, a phospholipids-rich dairy powder, could be used as an
emulsifier in different food systems.
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16. Material & Method
Three batches of whey cream, around 120 L each.
The product was derived from unsalted whey obtained from renneted
cheese processing.
The cream was processed at 12oC using a rotatory churn to obtain whey
butter and butter milk.
Butter milk was recovered in milk cans, after butter fines were removed by
filtration through cheese cloth, and stored overnight at 4oC until
membrane filtration.
A pilot plant scale system (R-12 model, GEA-Niro Filtration, Hudson,
WI,USA) using two spiral polymeric membranes fitted in parallel on the
module(10kDa molecular mass cut off,11.33m2 total surface area) was used
for butter milk concentration.
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17. Contd…
The process was carried out at 25oC, the transmembrane pressure was
around 6bar and feed pump was operated at 35Hz.
The ultrafiltration(UF) was conducted until a ten-fold volumetric
concentration factor was reached.
Diafiltration (DF) was done by adding continuously tap water at 25oC to
the feed tank to replace the removed permeate until reaching a five-fold
diafiltration factor (5XDF).
The final retentate from all experiments were spray-dried (Niro Filter lab
Spray-drier, Hudson, WI, USA) using 35 bar of pressure, and inlet and
outlet air temperatures of 185 C and95oC, respectively, to obtain whey
butter milk powders(WBP).
A portion of the powder obtained from each whey cream batch was
submitted to supercritical fluid extraction(SFE).
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18. Contd…
Circulated deionized water at 3oC was used for cooling different zones in the
SFE apparatus.
Carbon dioxide tanks (50-lb) were filled.
The system conditions were controlled manually by Windows 2000 based
software.
Approximately135g of each sample were submitted to three extraction cycles
using the following conditions: 1500g of CO2 at a flow rate of 20g min-1,
extraction pressure of 350bar, and both extraction and collection temperature
of 50oC. The SFE trials were done in triplicate.
The powders submitted to the SFE (SFE-WBP)as well as the WBP were stored
at10oC until further analysis.
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19. Results
Composition of whey cream, Butter &
BM Composition of WBM & SFE-WBP
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20. Contd…
The permeate flux during the filtration started at 45Lh-1m-2, dropped
approximately 26% in the ultrafiltration step and increased gradually in the
diafiltration phase, reaching 36Lh-1m-2 at the end of the process.
The average flux for the whole filtration process was 31Lh-1m-2.
The SFE process removed approximately 34 g of lipids from each 100 g of sample
(47.26 g of lipids), using 97 g of carbon dioxide to extract each gram of this fat.
This extraction represented a reduction of 72% in the quantity of total lipids
originally present in the whey buttermilk powder.
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21. Contd…
As expected, the extraction using
supercritical CO2 selectively removed
non-polar lipids from the matrix
powder.
The removed fat did not contain any
polar lipids, as can be seen in the
chromatogram(lines7 and 8),
consequently their concentration
increased in the powders after the SFE
procedure (lines 4-6) when compared
with the non- treated whey butter
milk powders (lines1-3).
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22. Contd…
SFE did not influence the proportion of the phospholipids in the powder, since this
process did not remove any of them.
However, the lipid to protein ratio was reduced from 1:1 to1:3.5 while the
phospholipid to lipid ratio increased from 1:6.6 to1:1.7.
The whole filtration (UF/DF) and SFE processing resulted in an increase of 500% in
the phospholipid content, in comparison to the original whey buttermilk on dry
matter basis.
The SDS-PAGE protein profiles revealed that all samples had three distinct protein
groups: milk fat globule membrane proteins, caseins and immunoglobulin G light
chains, and the main whey proteins β-Lactoglobulin and a-lactalbumin.
The samples of whey buttermilk, whey buttermilk powder, and whey buttermilk
powder after SFE showed a lower proportion of proteins in the molecular weight
range of caseins and Ig-G light chains and a greater proportion of MFGM and whey
proteins, especially after the SFE step.
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23. Downstream Processing of Bovine
Lactoferrin from Sweet Whey
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ARTICLE1I
ULBER R., PLATE K., WEISS T., DEMMER W., BUCHHOLZ H. AND SCHEPER T. (2001)
24. Introduction
Whey is a by-product from the cheese manufacturing process that is often
used to produce whey protein concentrate powders for food applications.
Besides the major whey proteins such as Lactalbumin or BSA, minor whey
proteins are present such as lactoperoxidase and bLF.
In addition to the well-known biological functions as an antimicrobial and
antiviral agent, bLF shows immunomodulatory functions in the host
defence system.
For the isolation of bLF, a two-step downstream process was developed
based on membrane systems.
The glycoprotein lactoferrin (LF) belongs to the transferrin protein family
with binding properties three times stronger to iron than to transferrin.
LF is also particularly involved in ALZHEIMER'S disease and could serve as
an HIV prevention clinical agent.
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25. Contd…
Only minor amounts in bovine milk ranging from 20–200 μg/ml.
Due to the basic isoelectric point (8.0–9.5) and the almost
positive charge, bLF can induce interactions with other whey and
milk proteins such as β-lactoglobulin and caseins.
To date, minor protein components like bLF are isolated by
standard column chromatographic procedures.
At present, Sartobind® adsorber (SARTORIUS, Göttingen,
Germany) is used for the direct removal of bLF from sweet whey.
Due to its high isoelectric point of 8.5–9.0, bLF can be bound on
the strong cationic membrane adsorber (Sartobind S®).
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26. Membrane Adsorber Technique
Membranes can be converted into efficient absorbers by attaching functional groups
to the inner surface of synthetic microporous membranes.
Depending on the choice of the attached functional groups, various adsorber systems
can be obtained that can be used for affinity adsorption, ion exchange or immobilized
metal affinity chromatography (IMAC).
Commercially available are membrane ion exchangers of the strongly acidic (sulfonic
acid), the strongly basic (quaternary ammonium), the weakly acidic (carboxylic acid)
and the weakly basic (diethylamine) type.
The membrane adsorber technology is typically applied for the concentration of
proteins and monoclonal antibodies, removal of contaminants (e.g. DNA, endotoxins)
and reduction of virus content.
Using such membrane adsorber technique requires a pre-treatment of whey to
remove the insoluble particles and lipids which otherwise will block the membrane.
continuous crossflow filtration steps have to be performed prior to the ion exchange
step. The permeate coming from this filtration step can be loaded directly on the
membrane adsorber.
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27. Major advantages of membrane adsorber
technology
Major advantages over classical separation methods:
Due to the membrane structure, diffusion processes do not limit the
binding of proteins; therefore, the loading and elution can be performed
at very high fluxes resulting in very short cycle times.
The compressibility of the membrane under normal operation conditions
can be neglected, channelling cannot occur, and the pressure distribution
inside the modules is designed to have plug flow through the module
altogether leading to sharp breakthrough curves.
Scale-up is very easy, materials and systems allow CIP and the validation
of the process is made easier due to the usage of standard products and
validation service of suppliers.
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28. Scheme of the downstream process for the isolation of bLF from
sweet whey
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29. Materials and Methods
1. Substances:
Sweet cheese whey.
Salts for sodium phosphate buffers (NaH2PO4 and Na2HPO4) and eluents (NaCl).
Bovine lactoferrin for standard measurements.
The matrix substance used for MALDI-MS measurements (matrix assisted laser
desorption/ ionization-mass spectrometry), α-cyano-cinnetatic acid, and the
chemicals for gel electrophoresis (sodium dodecyl sulphate,
ethylenediaminetetraacetic acid, bromphenol blue, tromethamine, sodium
thiosulfate, silver nitrate).
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30. Contd… 2. Crossflow Filtration: For the crossflow filtration of cheese whey, various membranes of
different geometries and pore sizes were used as shown in Table:
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31. Contd…
The experiments were carried out in the recycled batch mode, in
which the retentate as well as the permeate was led back into the
batch reactor to keep the concentration of bLF constant over the
whole filtration time.
To assess the cleaning effect of the filtrations, transmission
measurements were made against water at 600 nm (Uvikon 922
spectrophotometer, KONTRON INSTRUMENTS) with retentate and
permeate samples.
Permeates showing transmission values of at least 30 did not cause
blocking effects on the ion exchange membranes anymore.
The filtration temperature was 50 °C.
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32. 3. Cation-Exchange Membranes
For the isolation of bovine lactoferrin, strongly acidic membranes of the so-
called “Sartobind ® Membrane Adsorber Factor Two Family” from
SARTORIUS, Göttingen, Germany were used.
The reeled modules are available in various heights and layers, resulting in
membrane areas ranging from 0.1 to 8.0 m2 per module.
The presented results were received with two 1- m2 modules (type S-10k-15-
25) in series.
The flow rates varied between 1.0 and 3.0 l/min; the process was monitored
by an UV detector (model 662 UV Analyzer, WEDGEWOOD TECHNOL., Inc.) at
280 nm.
Prior to use, the membranes were pre-equilibrated with a 20 mM sodium
phosphate buffer at pH 7.0; afterwards bLF and lactoperoxidase as a by-
product were eluted in a three-step sodium chloride salt gradient.
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33. Contd…
4. Recovery:
The eluates were desalted and concentrated using a “Sartocon II” plant from SARTORIUS,
Göttingen, Germany.
A cellulose acetate membrane with an area of 0.7 m2 and a cut-off of 30 kDa was used.
The removal of sodium chloride was monitored via conductivity measurement.
The desalted lactoferrin solution was lyophilized.
5. Analysis:
The quantitative determination of bLF in solution was carried out using HPLC.
The elution gradient for the by-product lactoperoxidase was 0.5 M sodium chloride in 20 mM
sodium phosphate buffer (pH 7.0) and 2.0 M sodium chloride in 20 mM sodium phosphate
buffer (pH 7.0) for the target protein.
For the qualitative determination of bovine lactoferrin, MALDI-MS measurements with a
Compact Maldi 3 (KRATOS ANALYTICAL) and SDS-PAGE (sodium dodecyl sulphate-
polyacrylamide gel electrophoresis) with a PhastSystemTM (PHARMACIA, Uppsala, Sweden)
were used.
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34. Results and Discussion
The process for the production of bovine lactoferrin from cheese whey is divided into three
steps: a crossflow microfiltration, the isolation via cation-exchanger membranes and
recovery.
In the first step, the crossflow filtration, insoluble particles such as lipids, caseins and
precipitated proteins are removed from the whey. This is necessary to prevent blocking of
the cation exchange membrane modules in the isolation step.
To obtain suitable permeates in high yields with a maximum content of the target protein, a
membrane screening was carried out (The results of this screening are shown in Table).
The tubular modules allowed the highest permeate fluxes and constantly high permeation
rates of bovine lactoferrin to be obtained.
Based on their geometry, tubular modules also allow extremely high retentate fluxes to be
achieved; the resulting shear rates of about 9000 1/s prevent the fouling of the membrane's
surface, which may limit flux and permeation rates.
The pore size of 1 μm did not lead to permeates of adequate quality (transmission values of
less than 30), therefore, for the following whey filtrations a Microdyn 0.2 μm tubular module
was used.
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35. Contd…
In the second step, bLF was isolated from the permeates by using Sartobind® cation
exchanger membranes.
The dynamic binding capacity of the cation-exchange membranes for bovine lactoferrin was
found to be 0.2 mg/cm2 (Table) independent of the flow rate.
This allows 4 g of the target protein per process cycle to be isolated.
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36. Contd…
The dynamic binding capacity was determined at 10% breakthrough of bLF in the filtrate of
the adsorber step.
Figure below shows the bLF breakthrough curve for the used adsorber system (two modules
S-10k-15-25 in series).
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37. Contd…
After loading and rinsing the modules with 20 mM sodium phosphate buffer (pH
7.0) to remove non-bound components, a step gradient (0.2 –1 M NaCl) was used
for the elution of bLF and the second bound protein, lactoperoxidase.
90% of the bound lactoperoxidase was eluted from the modules at a salt
concentration of 0.2 M NaCl, together with 16% (0.64 g) of the bound lactoferrin.
The 0.3 M NaCl fraction contained 8% of lactoperoxidase and 48% (1.92 g) of
lactoferrin.
The 0.4 M NaCl fraction still contained rests of lactoperoxidase and 32% (1.28 g) of
the bound lactoferrin.
At salt concentrations of more than 0.5 M NaCl, no more proteins could be detected
in the eluates.
To get two protein fractions as pure as possible, the salt concentration of the used
eluents was optimized.
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38. Contd…
The best result was achieved with a three-step sodium chloride elution which led to a
lactoperoxidase fraction of about 85% purity (0.08 g bLF) and a lactoferrin fraction of about
95% purity (3.4 g bLF).
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39. Contd…
The content of lactoferrin in the processed cheese whey was 0.1 g/l.
Loading the modules with a flow rate of 2 l permeate per minute it was possible to
carry out two cycles per hour with a yield of 8 g bLF.
Several cycles were performed successively without a cleaning procedure so that
the loading step of each new process cycle could lead to a self-regeneration of the
ion-exchange groups on the surface of the modules.
The desalting and concentration of the protein-containing eluate was carried out
by crossflow ultrafiltration.
The collected lactoferrin-containing permeates of five process cycles of about 12 l,
were desalted within two hours.
The loss of protein that was bound to the membrane's surface was 10%.
By rinsing the module with distilled water after finishing the ultrafiltration, the loss
could be reduced to 6%.
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