This study evaluated the effects of storage conditions on the stability of six commercial phytase products over 360 days. Phytases were stored either in pure form, mixed with a vitamin premix, or mixed with a vitamin and trace mineral premix at temperatures of -18°C, 5°C, 23°C, or 37°C. Storage stability was reported as residual phytase activity compared to initial levels. Results showed that phytase stability was affected by storage duration, temperature, product form, coating, and phytase source. Pure products stored at 23°C or below retained the highest levels of activity. Coating helped mitigate losses at higher temperatures. Mixing phytases with premixes and
Optimization of key process variables for enhanced refamycin b production in ...ijabjournal
In the present study of solid media conditions for the refamycin B yield by solid state fermentation was studied and optimized using both classical method and statistical design of experiments). Statistical analysis of the results of Plackett–Burman showed that the lower level of initial moisture , initial pH, barbital, glucose and to solid media, or increase in the concentration of xylose in the range tested, results in significant effect in refamycin B yield of AmycolatopsisrifamycinicaMTCC 14 by solid state
fermentation. The effect of change in the levels of initial moisture, initial pH, barbital, glucose and xylose
on the rfefamycin B yield was studied using central composite design methodology. Statistical analysis of
the data showed that all the independent process had significant effect on refamycin B yield. The interaction between initial moisture and initial pH, between initial moisture and barbital, between initial moisture and glucose, between initial moisture and xylose, between initial pH and xylose, between barbital and glucose, between barbital and xylose, and between glucose and xylose were significant when the response was refamycin B.
Extraction, chemical composition, use in induced protection and cross-reactiv...IJEAB
Exopolysaccharides (PS) are the major components on the surface of bacteria and also produced by fungi. These molecules are important in human health, in order to control diabetes as well as protect plants against attacks of foliage diseases. The objective of the present work was to study the partial chemical structure of the carbohydrate, use in control disease in plants and cross-serological relationship (cross-reactive antigens between isolates from fungi (Tremella fuciformis (Tf) and bacteria (Xanthomonas campestris pv. citri (Xcc)). Tf was developed in culture medium containing sorghum seeds during 20 days, and Xcc in the PDA (potato dextrose agar) medium for an 8 days period. The polysaccharide was removed from the culture medium, precipitated with ethanol, and quantified total sugar. By TLC was observed that 2 isolates presented galactose, glucose, mannose, arabinose and xylose in different proportions. Fucose and ribose was not found in the PS from Xcc but present in Tf. In ELISA, antiserum to Xcc revealed an antigenic homologous reaction with the same bacteria and heterologous with Tf. Barley plants pretreated with PS from Tf and later challenged with conidia from B.sorokiniana, demonstrated protection against the pathogen. Results suggested that PS from Tf presented induction of protection. Both PS (antigens) present an identical epitope demonstrated by reaction in Elisa test. The antibody against Xcc was specific for an epitope and bounded to another antigen due to having similar chemical properties.
Development of a Bioactive Food Additive for Controlling of Fungal GrowthIJEAB
Fresh foods have a great importance in human nutrition. However, they are marketed with greatly reduced shelf life mainly due to fungal spoilage. In this work, cell wall degrading enzymes produced by Trichoderma asperellum T00 (TCWDE) were immobilized onto cashew gum polysaccharide (CGP) in order to evaluate the potential use of this material as food additive aiming to increase the shelf life by inhibiting fungal growth. Results from factorial design (32) evidenced that the best conditions for TCWDE immobilization was achieved with 20 min of reaction using 1 mmol L-1 of NaIO4. On these conditions it was observed 91% of retention yield for NAGase (30.1 ± 0.38 U mL-1), 41% for chitinase (0.67 ± 0.05 U mL-1), and 24% for β-1,3-glucanase (0.017± 0.001 U mL-1). CGP/TCWDE was effective for growth inhibition of Aspergillus fumigatus and Penicillium sp. and the inhibition mechanism seems to involve changes in the cell wall of those microorganisms. Finally, the CGP/TCWDE presented high stability after drying, maintaining enzymatic and biological activity after 200 days of storage at room temperature (25 ºC).
Optimization of key process variables for enhanced refamycin b production in ...ijabjournal
In the present study of solid media conditions for the refamycin B yield by solid state fermentation was studied and optimized using both classical method and statistical design of experiments). Statistical analysis of the results of Plackett–Burman showed that the lower level of initial moisture , initial pH, barbital, glucose and to solid media, or increase in the concentration of xylose in the range tested, results in significant effect in refamycin B yield of AmycolatopsisrifamycinicaMTCC 14 by solid state
fermentation. The effect of change in the levels of initial moisture, initial pH, barbital, glucose and xylose
on the rfefamycin B yield was studied using central composite design methodology. Statistical analysis of
the data showed that all the independent process had significant effect on refamycin B yield. The interaction between initial moisture and initial pH, between initial moisture and barbital, between initial moisture and glucose, between initial moisture and xylose, between initial pH and xylose, between barbital and glucose, between barbital and xylose, and between glucose and xylose were significant when the response was refamycin B.
Extraction, chemical composition, use in induced protection and cross-reactiv...IJEAB
Exopolysaccharides (PS) are the major components on the surface of bacteria and also produced by fungi. These molecules are important in human health, in order to control diabetes as well as protect plants against attacks of foliage diseases. The objective of the present work was to study the partial chemical structure of the carbohydrate, use in control disease in plants and cross-serological relationship (cross-reactive antigens between isolates from fungi (Tremella fuciformis (Tf) and bacteria (Xanthomonas campestris pv. citri (Xcc)). Tf was developed in culture medium containing sorghum seeds during 20 days, and Xcc in the PDA (potato dextrose agar) medium for an 8 days period. The polysaccharide was removed from the culture medium, precipitated with ethanol, and quantified total sugar. By TLC was observed that 2 isolates presented galactose, glucose, mannose, arabinose and xylose in different proportions. Fucose and ribose was not found in the PS from Xcc but present in Tf. In ELISA, antiserum to Xcc revealed an antigenic homologous reaction with the same bacteria and heterologous with Tf. Barley plants pretreated with PS from Tf and later challenged with conidia from B.sorokiniana, demonstrated protection against the pathogen. Results suggested that PS from Tf presented induction of protection. Both PS (antigens) present an identical epitope demonstrated by reaction in Elisa test. The antibody against Xcc was specific for an epitope and bounded to another antigen due to having similar chemical properties.
Development of a Bioactive Food Additive for Controlling of Fungal GrowthIJEAB
Fresh foods have a great importance in human nutrition. However, they are marketed with greatly reduced shelf life mainly due to fungal spoilage. In this work, cell wall degrading enzymes produced by Trichoderma asperellum T00 (TCWDE) were immobilized onto cashew gum polysaccharide (CGP) in order to evaluate the potential use of this material as food additive aiming to increase the shelf life by inhibiting fungal growth. Results from factorial design (32) evidenced that the best conditions for TCWDE immobilization was achieved with 20 min of reaction using 1 mmol L-1 of NaIO4. On these conditions it was observed 91% of retention yield for NAGase (30.1 ± 0.38 U mL-1), 41% for chitinase (0.67 ± 0.05 U mL-1), and 24% for β-1,3-glucanase (0.017± 0.001 U mL-1). CGP/TCWDE was effective for growth inhibition of Aspergillus fumigatus and Penicillium sp. and the inhibition mechanism seems to involve changes in the cell wall of those microorganisms. Finally, the CGP/TCWDE presented high stability after drying, maintaining enzymatic and biological activity after 200 days of storage at room temperature (25 ºC).
In recent years, aquaculture has gained in importance as a renewable source of dietary protein and as a viable commercial activity. To maintain this position in the future and to continue to provide a good investment opportunity, the problems the sector currently faces must be addressed. One of the more important of these concerns is the cost of feed, which is estimated to be 50-60 percent of the total cost of production. Numerous studies on the use of different feed formulations, feed ingredients and feeding techniques have been conducted (Kaushik et al., 2004; Thiessen et al., 2003; Martinez et al., 2004; Enes et al.,2006; Izquierdo et al., 2003). These studies have included assessments of various alternative raw materials, vitamins and minerals, monitoring the amount of feed provided to the fish, and the addition of pigments and other feed additives to the diet. In particular, various feed additives with growth promoting properties came into prominence in these studies (Francis et al., 2005; Haroun et al., 2006; Abdel-Tawwab et al., 2008; Lara-Flores et al., 2003; Li and Gatlin, 2004;). Growth promoting feed additives may contain different ingredients as plant extracts, organic acids, probiotics, hormones etc.
antimicrobial packaging a type of active packaging in which antimicrobial agents are added to a conventional packaging or it maybe a inheriant just like chitosan. its is considered third type of packaging to prevent microbial decay and hence enhance selflife of package
Synthesis, Characterization, and Biological Evaluation of Some Novel Phthalim...BRNSS Publication Hub
Objective: Different Phthalimide derivatives (4,5,6,7-tetrachloro-2-[1,2,4]triazol-4-yl]-isoindole-1,3-dione derivatives) were synthesized and biological activities of them were evaluated. Materials and Methods: In the present study, four new phthalimide derivatives were synthesized. The structures of final compounds were characterized on the basis of spectral data. Then, biological evaluation of all the synthesized compounds means in vivo anticancer activity was evaluated on the Ehrlich ascites carcinoma (EAC) bearing Swiss albino mice model, and in vitro antioxidant activity was assessed using 1,1-diphenyl-2-picryl hydrazine (DPPH) radical scavenging assay. Results: The titled compounds (2A-2D) were found to reduce tumor volume, viable cell count and increase non-viable cell count, and percentage increase in life span. All compounds showed significant activity in quenching DPPH free radical. Conclusion: All compounds showed significant (P<0.01) anticancer activity compared to EAC control group. All synthesized compound exhibited considerable antioxidant activity against free radicals, and they showed significant IC50 values and can, thus, ensure protection against oxidative stress.
Properties of biopolymers produced by transglutaminase treatment of wpi and g...Eduard Hernàndez I PMP®
Byproduct utilization is an important consideration in the development of sustainable processes. Whey
protein isolate (WPI), a byproduct of the cheese industry, and gelatin, a byproduct of the leather industry,
were reacted individually and in blends with microbial transglutaminase (mTGase) at pH 7.5 and 45 C.
When a WPI (10% w/w) solution was treated with mTGase (10 U/g) under reducing conditions, the viscosity
increased four-fold and the storage modulus (G0) from 0 to 300 Pa over 20 h. Similar treatment
of dilute gelatin solutions (0.5–3%) had little effect. Addition of gelatin to 10% WPI caused a synergistic
increase in both viscosity and G0 , with the formation of gels at concentrations greater than 1.5% added
gelatin. These results suggest that new biopolymers, with improved functionality, could be developed
by mTGase treatment of protein blends containing small amounts of gelatin with the less expensive whey
protein.
Feed ingredients and feed prices are increasing; it is becoming harder to maintain the nutritional balance of the feed without increasing too much the feed price. Now, the use of ingredients from less stringent quality is likely to increase. Though plant materials are usually more reasonable in price than animal products, they can present problems through the presence of naturally occurring contaminants. Indeed, contamination of feed commodities by microorganisms and mycotoxins is the first negative factor impacting animal feed quality. Numerous researches have studied the decrease of performances with contaminated feeds.
Microbial Production Of Alkaline Proteases And Evaluation Of Its Performances...Shafkat Shamim Rahman
A high alkaline protease producing bacterial strain was isolated and identified a local soil sample. The organism was gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus subtilis. It was also identified as B. subtilis with 99.9% identity by API 50 CHB. The enzyme hydrolyses a number of proteins including azocasein which suggests that it is an extracellular alkaline protease. The experimentally determined isoelectric point was 5.1 and the optimal enzyme activity was at 60°C and at pH 8.5. The esterase preferentially hydrolyzed short-chain fatty acids. Native enzyme preparations typically showed a Michaelis constant (Km) and Vmax of 0.40mM and 12,200 U mg)-1, respectively. This microbial enzyme was partially purified by ammonium sulfate fractionation, dialysis, DEAE cellulose chromatography and electrophoretic analysis. Enzyme purity was tested by SDS-PAGE. Quantitative estimation has shown that 40mL of culture supernatant could dehair 2×1 cm of leather completely in 9 hours. In future the tanneries will use a combination of chemical and enzymatic processes. In practical applications, protease is a useful enzyme for promoting the hydrolysis of proteins and showing significant industrial applications.
ABSTRACT- The present study was undertaken to make paneer enriched with fiber otherwise fiber deficient paneer. Coconut powder is in the form of fiber was included in the preparation of paneer. Paneer is one such product which is a regular dietary favorite among the Indians. Paneer has short life span at room temperature. So, the present study was aimed to assess the shelf life of salted paneer at different intervals in refrigeration temperature and physico-chemical attributes also. Paneer is prepared by combined action of acid coagulants and heat treatment of buffalo and cow milk or a combination thereof. Paneer have pleasant odour and characteristic mild acidic flavour. No extraneous coloring matter should be added to paneer at any stage. Paneer is a highly perishable product and has limited shelf life, largely because of its high moisture content. Its shelf life was reported to be only six days under refrigeration, though its freshness is lost within three days. The spoilage of paneer occurs mainly due to the growth of microorganisms, which bring about various physico-chemical changes. The growth of microorganisms can be delayed and shelf life of paneer be increased by addition of salt in the paneer. All treatment combinations were analyzed for a total viable count (bacteria) on nutrient agar and fungi on PDA and Coliform on Mcconkey agar. All the samples had bacteriological count ranging from 1x104 to 14x104 cfu/gm. And in all samples coliform was absent, so the product was found to be good and proper hygienic condition were maintain during the preparation, handling, and storage.
Key words: Paneer, Standard Plate Count, Chemical analysis, Yeast and mould count, Fiber
In recent years, aquaculture has gained in importance as a renewable source of dietary protein and as a viable commercial activity. To maintain this position in the future and to continue to provide a good investment opportunity, the problems the sector currently faces must be addressed. One of the more important of these concerns is the cost of feed, which is estimated to be 50-60 percent of the total cost of production. Numerous studies on the use of different feed formulations, feed ingredients and feeding techniques have been conducted (Kaushik et al., 2004; Thiessen et al., 2003; Martinez et al., 2004; Enes et al.,2006; Izquierdo et al., 2003). These studies have included assessments of various alternative raw materials, vitamins and minerals, monitoring the amount of feed provided to the fish, and the addition of pigments and other feed additives to the diet. In particular, various feed additives with growth promoting properties came into prominence in these studies (Francis et al., 2005; Haroun et al., 2006; Abdel-Tawwab et al., 2008; Lara-Flores et al., 2003; Li and Gatlin, 2004;). Growth promoting feed additives may contain different ingredients as plant extracts, organic acids, probiotics, hormones etc.
antimicrobial packaging a type of active packaging in which antimicrobial agents are added to a conventional packaging or it maybe a inheriant just like chitosan. its is considered third type of packaging to prevent microbial decay and hence enhance selflife of package
Synthesis, Characterization, and Biological Evaluation of Some Novel Phthalim...BRNSS Publication Hub
Objective: Different Phthalimide derivatives (4,5,6,7-tetrachloro-2-[1,2,4]triazol-4-yl]-isoindole-1,3-dione derivatives) were synthesized and biological activities of them were evaluated. Materials and Methods: In the present study, four new phthalimide derivatives were synthesized. The structures of final compounds were characterized on the basis of spectral data. Then, biological evaluation of all the synthesized compounds means in vivo anticancer activity was evaluated on the Ehrlich ascites carcinoma (EAC) bearing Swiss albino mice model, and in vitro antioxidant activity was assessed using 1,1-diphenyl-2-picryl hydrazine (DPPH) radical scavenging assay. Results: The titled compounds (2A-2D) were found to reduce tumor volume, viable cell count and increase non-viable cell count, and percentage increase in life span. All compounds showed significant activity in quenching DPPH free radical. Conclusion: All compounds showed significant (P<0.01) anticancer activity compared to EAC control group. All synthesized compound exhibited considerable antioxidant activity against free radicals, and they showed significant IC50 values and can, thus, ensure protection against oxidative stress.
Properties of biopolymers produced by transglutaminase treatment of wpi and g...Eduard Hernàndez I PMP®
Byproduct utilization is an important consideration in the development of sustainable processes. Whey
protein isolate (WPI), a byproduct of the cheese industry, and gelatin, a byproduct of the leather industry,
were reacted individually and in blends with microbial transglutaminase (mTGase) at pH 7.5 and 45 C.
When a WPI (10% w/w) solution was treated with mTGase (10 U/g) under reducing conditions, the viscosity
increased four-fold and the storage modulus (G0) from 0 to 300 Pa over 20 h. Similar treatment
of dilute gelatin solutions (0.5–3%) had little effect. Addition of gelatin to 10% WPI caused a synergistic
increase in both viscosity and G0 , with the formation of gels at concentrations greater than 1.5% added
gelatin. These results suggest that new biopolymers, with improved functionality, could be developed
by mTGase treatment of protein blends containing small amounts of gelatin with the less expensive whey
protein.
Feed ingredients and feed prices are increasing; it is becoming harder to maintain the nutritional balance of the feed without increasing too much the feed price. Now, the use of ingredients from less stringent quality is likely to increase. Though plant materials are usually more reasonable in price than animal products, they can present problems through the presence of naturally occurring contaminants. Indeed, contamination of feed commodities by microorganisms and mycotoxins is the first negative factor impacting animal feed quality. Numerous researches have studied the decrease of performances with contaminated feeds.
Microbial Production Of Alkaline Proteases And Evaluation Of Its Performances...Shafkat Shamim Rahman
A high alkaline protease producing bacterial strain was isolated and identified a local soil sample. The organism was gram positive and forms spore during adverse condition in the growth medium. After various tests it was suggested and the features agreed with the description of Bacillus subtilis. It was also identified as B. subtilis with 99.9% identity by API 50 CHB. The enzyme hydrolyses a number of proteins including azocasein which suggests that it is an extracellular alkaline protease. The experimentally determined isoelectric point was 5.1 and the optimal enzyme activity was at 60°C and at pH 8.5. The esterase preferentially hydrolyzed short-chain fatty acids. Native enzyme preparations typically showed a Michaelis constant (Km) and Vmax of 0.40mM and 12,200 U mg)-1, respectively. This microbial enzyme was partially purified by ammonium sulfate fractionation, dialysis, DEAE cellulose chromatography and electrophoretic analysis. Enzyme purity was tested by SDS-PAGE. Quantitative estimation has shown that 40mL of culture supernatant could dehair 2×1 cm of leather completely in 9 hours. In future the tanneries will use a combination of chemical and enzymatic processes. In practical applications, protease is a useful enzyme for promoting the hydrolysis of proteins and showing significant industrial applications.
ABSTRACT- The present study was undertaken to make paneer enriched with fiber otherwise fiber deficient paneer. Coconut powder is in the form of fiber was included in the preparation of paneer. Paneer is one such product which is a regular dietary favorite among the Indians. Paneer has short life span at room temperature. So, the present study was aimed to assess the shelf life of salted paneer at different intervals in refrigeration temperature and physico-chemical attributes also. Paneer is prepared by combined action of acid coagulants and heat treatment of buffalo and cow milk or a combination thereof. Paneer have pleasant odour and characteristic mild acidic flavour. No extraneous coloring matter should be added to paneer at any stage. Paneer is a highly perishable product and has limited shelf life, largely because of its high moisture content. Its shelf life was reported to be only six days under refrigeration, though its freshness is lost within three days. The spoilage of paneer occurs mainly due to the growth of microorganisms, which bring about various physico-chemical changes. The growth of microorganisms can be delayed and shelf life of paneer be increased by addition of salt in the paneer. All treatment combinations were analyzed for a total viable count (bacteria) on nutrient agar and fungi on PDA and Coliform on Mcconkey agar. All the samples had bacteriological count ranging from 1x104 to 14x104 cfu/gm. And in all samples coliform was absent, so the product was found to be good and proper hygienic condition were maintain during the preparation, handling, and storage.
Key words: Paneer, Standard Plate Count, Chemical analysis, Yeast and mould count, Fiber
Using next generation sequencing to describe epiphytic microbiota associated ...Agriculture Journal IJOEAR
Abstract— Its seems likely that agricultural management as diverse as certified organic and conventional IPM practices would impact the microbiota associated with crop surfaces differently. We sampled organic and conventionally managed apples at multiple time-points in a growing season and characterized the bacterial taxa associated with replicates of each treatment type. Surprisingly, no evidence of significant differences persisting across multiple time-points was observed. Significant differential abundance of certain taxa was documented but when it was, it was primarily associated with a single time-point making it difficult to understand if these observations resulted from an environmental or a treatment effect. Principal component analyses demonstrated that sampling time-point explained more of the differences between bacterial communities than treatment. Description of dominant shared bacterial families for both organic and conventional samples included Oxalobacteraceae, Pseudomonadaceae, Sphingomonadaceae and Enterobacteriaceae.
ABSTRACT- Live microorganisms, have beneficial effects on their host’s health, are called as probiotics. There are various possible sources to isolate
these bacteria. In this studyp harmaceutical probiotic sachet is used as isolation source. The purpose of this study is to search the potentiality of
probiotic bacteria and investigate the probiotic properties of isolates.9 different samples of 3 brands of sachet were used for isolation of bacteria.
Isolates were examined according to their probiotic properties. The probiotic characteristics like pH and Bile tolerance, Antagonistic activity and
Antibiotic susceptibility of isolated bacteria Such as Lactobacillus acidophilus, Lactobacillus rhamnosus and Bifidobacterium bifidum was done. Bile
Tolerance and pH tolerance was determined with the help of the help of coefficient of growth inhibition if their coefficient of growth inhibition is less
than 0.5 the organism was considered as the pH and Bile tolerance. The Strains of Lactobacillus acidophilus and Lactobacillus rhamnosus and Bifidobacterium
bifidum show best result at the pH Acidic to Neutral (5 to 7) and show a bile tolerance from 1-4 % bile. All the isolated bacteria show
the maximum inhibition against Staphyloccocus aureus and minimum against Salmonella typhi by Lactobacillus Strains but Bifidobacterium show
minimum against Escheria coli. Most isolates show resistance toward antibiotics. From this study it can be concluded that pharmaceutical probiotic
products used in the study were showing satisfactory quality and potential probiotic strain.
Key words- Probiotic, Lactobacillus, Bifidobacterium, Sachet
Phthalates, an alkyl aryl esters of 1, 2 benzenedicarboxylic acids are widely used as plasticizer. They show low water solubility, high oil solubility, high octanol-to-water partition coefficient and low volatility. Phthalates are easily released into the packed food because there is no covalent bond between the phthalates and plastics in which they are mixed. Phthalates may cause major ill effects like endocrine disruption, asthma and cancer. A study was conducted on tea (aromatic beverage), sambar (an Indian dish) and alcohol packed in polythene covers and plastic cups used in routine hotels and beverage shops. The samples were analyzed for plastic residues through gas chromatography coupled with mass spectrometry (GC-MS). The result shows that, the tea filled in polythene cover and alcohol filled in plastic cup did not show any leachate of phthalates. However, the sambar packed in the polythene cover showed leachate of phthalates. The diethylhexyl phthalate was observed to be 20% of Total Ion Chromatogram in sambar. Future studies can be carried out to identify the leaching effect of phthalates in different plastic packed food materials under different conditions.
Effects of dietary thyme essential oil on hemato-immunological indices, intes...Inacio Mateus Assane
Thyme (Thymus vulgaris) essential oil (TVEO) is a herbal medicine with one of the highest levels of antimicrobial activity. Although TVEO has been broadly used in poultry and swine production due to its immunostimulatory and growth-promoting characteristics, the effects of TVEO on fish are poorly characterized. In this study, Nile tilapia (Oreochromis niloticus L.) were fed 0, 0.1, 0.5, and 1% TVEO for 15 days. Subsequently, blood parameters, intestinal morphology, and the population of Bacillus bacteria in the intestine were evaluated. The numbers of lymphocytes (p < 0.05) and leukocytes (p < 0.05) significantly increased in the blood of the fish fed the highest dose of TVEO. Based on the normal behavior of the fish and the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, which were not altered (p > 0.05), this study concluded that the diets were safe and showed no negative or toxic effects. Even at doses as high as 1%, TVEO did not alter the population of beneficial Bacillus bacteria in the gut. In conclusion, supplementation with TVEO stimulated the cellular components of the non-specific immune response of Nile tilapia without causing deleterious effects or altering the population of important intestinal bacteria.
Penelitian berkaitan dengan pengurangan intensitas cahaya akibat penutupan awan sbg bentuk pengaruh dari perubahan iklim thd kualitas benih dan perkecambahan kedelai
Ang Chong Yi Navigating Singaporean Flavors: A Journey from Cultural Heritage...Ang Chong Yi
In the heart of Singapore, where tradition meets modernity, He embarks on a culinary adventure that transcends borders. His mission? Ang Chong Yi Exploring the Cultural Heritage and Identity in Singaporean Cuisine. To explore the rich tapestry of flavours that define Singaporean cuisine while embracing innovative plant-based approaches. Join us as we follow his footsteps through bustling markets, hidden hawker stalls, and vibrant street corners.
Hamdard Laboratories (India), is a Unani pharmaceutical company in India (following the independence of India from Britain, "Hamdard" Unani branches were established in Bangladesh (erstwhile East Pakistan) and Pakistan). It was established in 1906 by Hakeem Hafiz Abdul Majeed in Delhi, and became
a waqf (non-profitable trust) in 1948. It is associated with Hamdard Foundation, a charitable educational trust.
Hamdard' is a compound word derived from Persian, which combines the words 'hum' (used in the sense of 'companion') and 'dard' (meaning 'pain'). 'Hamdard' thus means 'a companion in pain' and 'sympathizer in suffering'.
The goals of Hamdard were lofty; easing the suffering of the sick with healing herbs. With a simple tenet that no one has ever become poor by giving, Hakeem Abdul Majeed let the whole world find compassion in him.
They had always maintained that working in old, traditional ways would not be entirely fruitful. A broader outlook was essential for a continued and meaningful existence. their effective team at Hamdard helped the system gain its pride of place and thus they made an entry into an expansive world of discovery and research.
Hamdard Laboratories was founded in 1906 in Delhi by Hakeem Hafiz Abdul Majeed and Ansarullah Tabani, a Unani practitioner. The name Hamdard means "companion in suffering" in Urdu language.(itself borrowed from Persian) Hakim Hafiz Abdul Majeed was born in Pilibhit City UP, India in 1883 to Sheikh Rahim Bakhsh. He is said to have learnt the complete Quran Sharif by heart. He also studied the origin of Urdu and Persian languages. Subsequently, he acquired the highest degree in the unani system of medicine.
Hakim Hafiz Abdul Majeed got in touch with Hakim Zamal Khan, who had a keen interest in herbs and was famous for identifying medicinal plants. Having consulted with his wife, Abdul Majeed set up a herbal shop at Hauz Qazi in Delhi in 1906 and started to produce herbal medicine there. In 1920 the small herbal shop turned into a full-fledged production house.
Hamdard Foundation was created in 1964 to disburse the profits of the company to promote the interests of the society. All the profits of the company go to the foundation.
After Abdul Majeed's death, his son Hakeem Abdul Hameed took over the administration of Hamdard Laboratories at the age of fourteen.
Even with humble beginnings, the goals of Hamdard were lofty; easing the suffering of the sick with healing herbs. With a simple tenet that no one has ever become poor by giving, Hakeem Abdul Majeed let the whole world find compassion in him. Unfortunately, he passed away quite early but his wife, Rabia Begum, with the support of her son, Hakeem Abdul Hameed, not only kept the institution in existence but also expanded it. As he grew up, Hakeem Abdul Hameed took on all responsibilities. After helping with his younger brother's upbringing and education, he included him in running the institution. Both brothers Hakeem Abdul Hameed and Hakim Mohammed
Roti Bank Hyderabad: A Beacon of Hope and NourishmentRoti Bank
One of the top cities of India, Hyderabad is the capital of Telangana and home to some of the biggest companies. But the other aspect of the city is a huge chunk of population that is even deprived of the food and shelter. There are many people in Hyderabad that are not having access to
Vietnam Mushroom Market Growth, Demand and Challenges of the Key Industry Pla...IMARC Group
The Vietnam mushroom market size is projected to exhibit a growth rate (CAGR) of 6.52% during 2024-2032.
More Info:- https://www.imarcgroup.com/vietnam-mushroom-market
2. protein, phytases lose a significant amount of activity
when subjected to feed processing treatments (Jong-
bloed and Kemme, 1990; Spring et al., 1996); thus, re-
search has focused on optimizing the thermostability of
exogenous phytases in industrial settings. Nutritionists
most often consider the minimum guaranteed phytase
after feed processing in their diet formulations; how-
ever, the stability of phytases during storage receives
little attention. In some circumstances, because of
poor inventory management, we have observed classi-
cal signs of rickets in pigs because of extended phytase
storage of 6 to 12 mo. No independent study has been
conducted to evaluate the effects of various factors such
as coating, time, or storage temperature on the stabil-
ity of commercial phytases. Some suggest that poten-
tial interactions may occur between phytase and some
components of the premixes and that such interaction
may affect phytase activity; however, no data support
this assumption. Therefore, the objective of this study
was to determine the effects of coating, storage form,
storage temperature, and duration of storage on the
stability of 6 commercially available phytases.
MATERIALS AND METHODS
This study was conducted at the Animal Nutrition
Laboratory and at the Bioprocessing and Industrial
Value Added Program Building at Kansas State Uni-
versity (Manhattan, KS). Animal Care and Use Com-
mittee approval was not obtained for this experiment
because no animals were used.
Phytase Sources
This experiment used 6 commercially available phy-
tases: OptiPhos 2000-M [uncoated, declared potency of
2,000,000 phytase units (FTU)/kg; Phytex LLC, Sher-
idan, IN], OptiPhos 2000-PF (coated, declared poten-
cy of 2,000,000 FTU/kg; Phytex LLC), Phyzyme XP
5000 G (uncoated, declared potency of 5,000,000 FTU/
kg; Danisco Animal Nutrition, Marlborough, UK),
Phyzyme XP 10000 TPT (coated, declared potency of
10,000,000 FTU/kg; Danisco Animal Nutrition), Rono-
zyme P-M [uncoated, declared potency of 50,000,000
phytase units (FYT)/kg; DSM Nutritional Products,
Basel, Switzerland], and Ronozyme P-CT (coated,
declared potency of 10,000,000 FYT/kg; DSM Nutri-
tional Products). One phytase unit (FTU or FYT) was
defined as the amount of enzyme that catalyzes the
release of 1 µmol of inorganic P per minute from 5.1
mM sodium phytate in pH 5.5 buffer at 37°C. Pure
cornstarch was used as a negative control because of
its minimal inherent phytase activity. The coated and
uncoated phytases were obtained from a third-party
distributor. For coated sources, a lipid or carbohydrate
coating encapsulates the phytase to minimize oxida-
tion at high temperatures, particularly during pelleting
conditions. The type of coating varies by manufacturer,
and its components are typically guarded as intellectual
property. The manufacturing dates of all products were
obtained from the original suppliers to ensure that the
products were within 6 mo of manufacture and were
not expired.
Pure Products
On d 0, 1.36 kg of each of the pure phytase products
and cornstarch were individually placed into 12 open,
single-lined paper bags. Three bags (observations) of
each product were stored in a freezer (−18°C), in a
refrigerator (5°C), at room temperature (23°C), and
in a controlled environment chamber set at 37°C and
75% humidity. At sampling, the bag was mixed and a
sample of approximately 50 g was taken. Each bag was
blind sampled at d 30, 60, 90, 120, 180, 270, and 360,
and samples were sent immediately after collection to
Technical Marketing Analytical Services of DSM Nutri-
tional Products Inc. (Belvidere, NJ) for phytase analy-
sis using a slight modification of AOAC official method
2000.12 (Engelen et al., 1994, 2001; AOAC, 2000). Each
bag of the cornstarch control, OptiPhos 2000-M, and
OptiPhos 2000-PF was blind sampled a second time,
and samples were sent to Phytex LLC (Portland, ME)
for phytase analysis using the Phytex method (Han
et al., 1999). Sampling of the pure products stored at
−18 and 5°C was discontinued after d 120 because not
enough material was available for sampling at −18°C
and because of mold growth from the high humidity
in the retained 5°C samples. Thus, only pure products
stored at 23 and 37°C were sampled for all time points.
Premixes
Each phytase product and the cornstarch control
were added and mixed with either the Kansas State
University vitamin premix or the Kansas State Univer-
sity vitamin and trace mineral (VTM) premix (Table
1). The amount added for each phytase product was de-
termined such that including 0.30% premix in the diet
would provide the activity of phytase recommended by
their respective manufacturers (250 FTU/kg, OptiPhos
2000-M and OptiPhos 2000-PF; 500 FTU/kg, Phyzyme
XP 5000 G and Phyzyme XP 10000 TPT; 1,850 FYT/
kg, Ronozyme P-M and Ronozyme P-CT).
A total of 2.46, 2.46, 1.36, 0.68, 2.52, and 0.50 kg of
pure OptiPhos 2000-M, OptiPhos 2000-PF, Phyzyme
XP 5000 G, Phyzyme XP 10000 TPT, Ronozyme
P-M, and Ronozyme P-CT, respectively, were weighed.
Cornstarch was added to the pure phytase products
to create 9.83 kg (OptiPhos) or 6.80 kg (Phyzyme XP
and Ronozyme P) batches, which were mixed using a
paddle mixer for 5 min. A total of 49.1 kg (OptiPhos)
or 34.0 kg (Phyzyme XP and Ronozyme P) of vitamin
or VTM premix was added to each batch and mixed
using a paddle mixer for an additional 12 min to cre-
ate premix batches of 59.0 kg (OptiPhos) or 40.8 kg
4263Storage stability of phytase
3. (Phyzyme XP and Ronozyme P). Additionally, 59.0 kg
of cornstarch made up the control batch.
The 7 batches each were divided equally into 6 open,
single-lined paper bags. Three bags of each batch were
stored either at room temperature (approximately 23°C
and <40% humidity) or in the environmentally con-
trolled chamber set at 37°C and 75% humidity. Before
sampling, each bag was mixed to ensure that a rep-
resentative sample was collected. A sample from each
bag was taken every 30 d until d 180, with 2 remain-
ing samples taken at d 270 and 360. Each sample was
blinded and sent immediately after collection to the
same laboratories as used for the pure samples for phy-
tase analysis.
Statistical Analyses
Data were analyzed using a mixed model (MIXED
procedure; SAS Inst. Inc., Cary, NC) to determine the
interactive and main effects of coating, storage form,
storage temperature, and time on the activity of 6 com-
mercially available phytase sources. Because the vi-
tamin and VTM premixes were stored only at room
temperature and in the environmentally controlled heat
chamber, 2 analyses were performed. The first was with
the pure forms only, and the second was for the pure
forms, vitamin premixes, and VTM premixes at 23 and
37°C. Least squares means were calculated for each in-
dependent variable. When treatment effect was a sig-
nificant source of variation, differences were determined
by using the preplanned pairwise comparisons (PDIFF
option of SAS). Statistical significance and tendencies
were set at P ≤ 0.05 and P < 0.10, respectively, for all
statistical tests.
RESULTS
Initial Phytase Activity
The calculated and analyzed initial (d 0) phytase
activities of the samples are shown in Table 2. On
the basis of the AOAC assay, the control samples for
the pure product, the vitamin premix, and the VTM
premix contained 4,967 to 10,500 phytase units/kg.
However, the phytase activities of the control samples
analyzed using the Phytex assay were much greater
than those analyzed using the AOAC assay. For all 3
products, the AOAC-analyzed phytase activities of Op-
tiPhos, Phyzyme XP, and Ronozyme P were 1.96 to
2.95, 0.97 to 1.57, and 1.03 to 1.42 times greater than
their calculated phytase activities, respectively. On the
basis of the Phytex assay, samples of the pure Opti-
Phos 2000-M and OptiPhos 2000-PF had similar (1.01
to 1.02 times) phytase activities compared with their
calculated activities. In contrast, the phytase activities
of both OptiPhos products added to the vitamin and
the VTM premix were less, ranging from 0.30 to 0.68 of
their calculated activities.
Pure Products
All interactive and main effects of phytase product,
coating, time, and storage temperature were significant
(P < 0.01; Table 3), except for the time × coating in-
teraction.
When the phytases were stored at 23°C or less, their
retained activity when stored in pure form decreased
(P < 0.01) as the duration of storage increased regard-
less of the phytase source or coating (Figures 1, 2, and
Table 1. Composition of the vitamin and vitamin and trace mineral (VTM) premixes
used in the study1
Item
Added per kilogram of
vitamin premix
Added per kilogram of
VTM premix
Vitamin
Vitamin A, IU 3,674,371 1,837,186
Vitamin D, IU 459,296 229,648
Vitamin E, IU 14,697 7,349
Vitamin K, mg 1,470 735
Riboflavin, mg 2,756 1,378
Niacin, mg 16,535 8,267
Pantothenic acid, mg 9,186 4,593
Cobalamin, mg 13 6
Trace mineral
Copper (CuSO4), mg — 4,593
Iodine [Ca(IO3)2], mg — 83
Iron (FeSO4), mg — 45,930
Manganese (MnSO4), mg — 11,023
Selenium (NaSeO3), mg — 83
Zinc (ZnSO4), mg — 45,930
1
The amount added for each phytase product was determined such that including 0.30% premix in the diet
would provide the phytase recommended by their respective manufacturers [250 phytase units (FTU)/kg,
OptiPhos 2000-M and OptiPhos 2000-PF (Phytex LLC, Sheridan, IN); 500 FTU/kg, Phyzyme XP 5000 G
and Phyzyme XP 10000 TPT (Danisco Animal Nutrition, Marlborough, UK); 1,850 phytase units (FYT)/kg,
Ronozyme P-M and Ronozyme P-CT (DSM Nutritional Products, Basel, Switzerland)].
4264 Sulabo et al.
4. 3). At d 30, 60, and 90, pure phytases retained at least
91, 85, and 78% of their initial phytase activity, respec-
tively. Until d 120, the pure forms retained 71 to 102%
of their initial phytase activity, except for uncoated
Ronozyme P-M, which retained 59% at 5°C. However,
storing pure products at 37°C had greater (P < 0.01)
effects on phytase stability. At d 30, both coated and
uncoated OptiPhos products stored in pure form re-
tained 91 to 93% of their initial activity when stored
at 37°C, whereas the coated and uncoated Phyzyme
phytases retained 69 to 74%. Coated Ronozyme P-CT
retained 69% of its initial phytase activity at d 30, but
uncoated Ronozyme P-M retained only 36%. After-
ward, phytases stored in pure forms retained at least
44, 39, and 33% of their initial phytase activities at
d 60, 90, and 120, respectively, except for uncoated
Ronozyme P-M, which retained only 5% of its initial
phytase activity at d 120. At d 180, 270, and 360, phy-
tases stored in pure forms at 37°C had retained phytase
activities ranging from 1 to 53%, compared with 50 to
109% when stored at 23°C.
Coated OptiPhos 2000-PF had similar retention
rates compared with uncoated OptiPhos 2000-M at d
Table 2. Calculated and analyzed phytase composition of samples at d 01
Item
Phytase composition
Calculated,
PU2
/kg
AOAC analysis,
PU/kg
AOAC
ratio3
Phytex analysis,
PU/kg
Phytex
ratio4
Pure product
Control5
0 10,500 — 3,343,000 —
OptiPhos 2000-M6
2,000,000 3,932,000 1.96 2,046,000 1.02
OptiPhos 2000-PF6,7
2,000,000 5,179,000 2.58 2,022,000 1.01
Phyzyme XP 5000 G8
5,000,000 5,144,000 1.03 — —
Phyzyme XP 10000 TPT7,8
10,000,000 10,587,000 1.06 — —
Ronozyme P-M9
50,000,000 52,148,500 1.04 — —
Ronozyme P-CT7,9
10,000,000 12,057,500 1.20 — —
Vitamin premix
Control5
0 4,967 — 37,000 —
OptiPhos 2000-M6
83,333 214,425 2.51 41,000 0.49
OptiPhos 2000-PF6,7
83,333 250,853 2.95 57,000 0.68
Phyzyme XP 5000 G8
166,666 266,339 1.57 — —
Phyzyme XP 10000 TPT7,8
166,666 266,116 1.57 — —
Ronozyme P-M9
616,666 738,388 1.19 — —
Ronozyme P-CT7,9
616,666 637,467 1.42 — —
Vitamin and trace mineral premix
Control5
0 4,948 — 77,000 —
OptiPhos 2000-M6
83,333 209,424 2.45 25,000 0.30
OptiPhos 2000-PF6,7
83,333 244,067 2.87 55,000 0.66
Phyzyme XP 5000 G8
166,666 209,437 1.23 — —
Phyzyme XP 10000 TPT7,8
166,666 166,239 0.97 — —
Ronozyme P-M9
616,666 699,542 1.13 — —
Ronozyme P-CT7,9
616,666 877,884 1.03 — —
1
Values represent means of 3 replicates sampled in duplicate. The AOAC analysis was performed at the DSM Nutritional Products laboratory
(Belvidere, NJ); the Phytex analysis was performed at Phytex LLC (Sheridan, IN).
2
PU = phytase units.
3
Ratio of average AOAC analyzed values to calculated values.
4
Ratio of Phytex analyzed values to calculated values.
5
Cornstarch used as the negative control.
6
Phytex LLC.
7
Coated phytase.
8
Danisco Animal Nutrition, Marlborough, UK.
9
DSM Nutritional Products, Basel, Switzerland.
Table 3. Probabilities of interactive and main effects
of storage time, temperature, coating, and phytase
product on stability (as defined by percentage of ini-
tial phytase activity) of commercially available phytase
products in pure forms
Item P-value
Interactive effect
Time × temperature × coating × product <0.001
Time × temperature × product <0.001
Time × temperature × coating <0.001
Time × coating × product <0.001
Temperature × coating × product <0.001
Temperature × coating <0.001
Temperature × product <0.001
Time × temperature <0.001
Time × coating 0.43
Time × product <0.001
Coating × product <0.001
Main effect
Time <0.001
Temperature <0.001
Coating <0.001
Product <0.001
4265Storage stability of phytase
5. 30 and 60 when stored at 37°C, but coating improved
(P < 0.01) retention rates from d 90 onward. Coat-
ing also improved (P < 0.01) the retained phytase ac-
tivities of Ronozyme phytase throughout the duration
of the study; however, coated Phyzyme had less (P <
0.01) phytase activity than uncoated Phyzyme until d
360. Among the coated phytases, the retention rates of
Ronozyme P-CT were less (P < 0.01) than those of Op-
tiPhos 2000-PF until d 120, whereas the retention rates
of Ronozyme P-CT were similar to those of Phyzyme
10000 TPT until d 90. Among the uncoated phytases,
OptiPhos 2000-M had greater (P < 0.01) phytase activ-
ity than both Phyzyme 5000 G and Ronozyme P-M at
d 30, but Phyzyme 5000 G retained more (P < 0.01)
phytase activity than the other 2 uncoated phytases
from d 90 onward.
Premixes
All interactive and main effects of phytase product,
form, coating, time, and storage temperature were sig-
nificant (P < 0.01; Table 4), except for the time × form
× coating and coating × temperature interactions (P
< 0.08).
When stored at 23°C, pure uncoated forms of Opti-
Phos 2000-M and Phyzyme 5000 G retained more (P
< 0.01) phytase activity with an increasing duration
of storage than did the phytase-supplemented vitamin
or VTM premixes (Figures 4, 5, and 6). However, the
phytase activity of pure, uncoated Ronozyme P-M was
similar among the pure, vitamin, and VTM samples.
Pure phytase products retained at least 85 and 72% of
Figure 1. Residual phytase activity (% of initial) for OptiPhos
2000-PF (coated; Phytex LLC, Sheridan, IN) and OptiPhos 2000-M
(uncoated; Phytex LLC) as affected by storage temperature [freezer
(−18°C), refrigerator (5°C), room temperature (23°C), and controlled
environment chamber (37°C and 75% humidity)] and time (30 to 120
d). Each data point (least squares means ± 2.32) is the mean of 3
observations.
Figure 2. Residual phytase activity (% of initial) for Phyzyme XP
10000 TPT (coated; Danisco Animal Nutrition, Marlborough, UK)
and Phyzyme XP 5000 G (uncoated; Danisco Animal Nutrition) as
affected by storage temperature [freezer (−18°C), refrigerator (5°C),
room temperature (23°C), and controlled environment chamber (37°C
and 75% humidity)] and time (30 to 120 d). Each data point (least
squares means ± 2.32) is the mean of 3 observations.
4266 Sulabo et al.
6. their initial phytase activities until d 180 and 360, re-
spectively, except for uncoated Ronozyme P-M (50%).
In contrast, phytase-supplemented vitamin premixes
retained at least 73% of their activity until d 180, ex-
cept for uncoated Phyzyme 5000 G (67%). At d 270
and 360, uncoated Phyzyme 5000 G and Ronozyme
P-M retained 56 to 59% of their initial phytase activi-
ties, whereas the rest of the phytases retained at least
68%. Among all the phytases, coated OptiPhos 2000-
PF retained the most activity (>92%; P < 0.01) until
d 360 when mixed with the vitamin premixes. In com-
parison, coated Ronozyme P-CT retained at least 83%
of its initial phytase activity, whereas coated Phyzyme
10000 TPT retained at least 73% of its activity until
d 360. For the phytase-supplemented VTM premixes,
retained phytase activities were at least 60% until d
180, except for uncoated OptiPhos 2000-M (43%). At d
270 and 360, uncoated OptiPhos 2000-M had only 28%
of its initial phytase activity, compared with at least
52% for the rest of phytases when mixed into the VTM
premixes. As in the vitamin premixes, coated OptiPhos
2000-PF retained the most activity (P < 0.01) among
all the phytases when mixed into the VTM premix-
es; however, its retention rates were less (P < 0.01)
than the rates obtained in the vitamin premixes. At d
360, coated OptiPhos 2000-PF, Ronozyme P-CT, and
Phyzyme 10000 TPT retained at least 83, 75, and 63%
of their initial phytase activities, respectively.
When stored at 37°C, retained phytase activities
were much less (P < 0.01) than the retention rates
observed in samples stored at 23°C regardless of the
phytase source, coating, or form of storage. For the
phytase-supplemented vitamin and VTM premixes, re-
tained phytase activities after only 30 d of storage were
59 and 62% on average, which were less (P < 0.01)
than the 72% for the pure phytase products. Uncoated
Ronozyme P-M was the least stable when mixed into
Figure 3. Residual phytase activity (% of initial) for Ronozyme
P-CT (coated; DSM Nutritional Products, Basel, Switzerland) and
Ronozyme P-M (uncoated; DSM Nutritional Products) as affected by
storage temperature [freezer (−18°C), refrigerator (5°C), room tem-
perature (23°C), and controlled environment chamber (37°C and 75%
humidity)] and time (30 to 120 d). Each data point (least squares
means ± 2.32) is the mean of 3 observations.
Table 4. Probabilities of interactive and main effects
of storage time, form, temperature, coating, and phy-
tase product on stability (as defined by percentage of
initial phytase activity) of commercially available phy-
tase products
Item P-value
Interactive effect
Time × form × coating × product × temperature <0.001
Time × form × coating × product <0.001
Time × form × coating × temperature <0.001
Time × form × product × temperature <0.001
Time × coating × product × temperature <0.001
Form × coating × product × temperature <0.001
Time × form × coating 0.07
Time × form × product <0.001
Time × form × temperature <0.001
Time × coating × product <0.001
Time × coating × temperature <0.001
Time × product × temperature 0.003
Form × coating × product <0.001
Form × coating × temperature 0.004
Form × product × temperature <0.001
Coating × product × temperature <0.001
Time × form <0.001
Time × coating 0.003
Time × product <0.001
Time × temperature <0.001
Form × coating <0.001
Form × product <0.001
Form × temperature <0.001
Coating × product <0.001
Coating × temperature 0.08
Product × temperature <0.001
Main effect
Time <0.001
Form <0.001
Coating <0.001
Product <0.001
Temperature <0.001
4267Storage stability of phytase
7. vitamin premixes, retaining only 31% of its initial phy-
tase activity at d 30. For the VTM premixes, uncoated
OptiPhos 2000-M was the most affected; it retained
only 20% of its initial phytase activity after a month
of storage. At d 180, the phytase treatments had 3 to
53% of their initial phytase activities. At the end of the
study (d 360), all the phytases had less than 28% of
their initial phytase activities.
The coated phytases stored in pure form or as phy-
tase-supplemented vitamin or VTM premixes had
greater (P < 0.01) phytase activities than the uncoated
phytases at all sampling periods. However, the differ-
ences in phytase activity between the coated and un-
coated phytases were smaller (P < 0.01) when they
were stored in pure forms than when the phytases were
in the vitamin and VTM premixes. At d 30, 60, and 90,
the differences in retained phytase activities between
the coated and uncoated phytases ranged from 4.2 to
4.5, 11.5 to 28.6, and 33.4 to 44% when the phytases
were in their pure form or as vitamin VTM premixes.
At d 30, coated phytases had similar phytase activities
among the 3 sources when stored at 37°C; however, un-
coated OptiPhos 2000-M and Phyzyme 5,000 XP phy-
tases stored in pure form had greater (P < 0.01) phy-
tase activities than those mixed with the vitamin and
VTM premixes. Likewise, uncoated OptiPhos 2000-M
retained greater (P < 0.01) phytase activity in vitamin
premixes than in VTM premixes.
DISCUSSION
Phytase Assays
Jones et al. (2010) previously demonstrated that ac-
curacy of the analysis for phytase activity depended
on the phytase product and the assay method. On the
Figure 4. Residual phytase activity (% of initial) for OptiPhos
2000-PF (coated; Phytex LLC, Sheridan, IN) and OptiPhos 2000-M
(uncoated; Phytex LLC) as affected by form of storage [as pure prod-
uct (PUR), in a vitamin premix (VIT), or in a vitamin and trace min-
eral premix (VTM)], storage temperature [room temperature (23°C)
and controlled environment chamber (37°C and 75% humidity)], and
time (30 to 360 d). Each data point (least squares means ± 3.75) is
the mean of 3 observations.
Figure 5. Residual phytase activity (% of initial) for Phyzyme XP
10000 TPT (coated; Danisco Animal Nutrition, Marlborough, UK)
and Phyzyme XP 5000 G (uncoated; Danisco Animal Nutrition) as
affected by form of storage [as pure product (PUR), in a vitamin
premix (VIT), or in a vitamin and trace mineral premix (VTM)],
storage temperature [room temperature (23°C) and controlled environ-
ment chamber (37°C and 75% humidity)], and time (30 to 360 d). Each
data point (least squares means ± 3.75) is the mean of 3 observations.
4268 Sulabo et al.
8. basis of the AOAC method, the initial phytase activity
of OptiPhos was 2 to 3 times greater than the calcu-
lated activity, which is similar (2.5 times) to the dif-
ference observed by Jones et al. (2010). The analyzed
initial phytase activities for Phyzyme and Ronozyme
were closer (1 to 1.6 times greater) to the calculated
activities, which was expected because the AOAC as-
say is the recommended method of analysis for these
products. For OptiPhos, the analyzed initial phytase
activity was similar to the calculated activity when the
Phytex assay (the recommended assay for this product)
was used. Therefore, these results confirmed our earlier
observations.
Phytases in Pure Forms
As with all catalytic proteins, phytases are sensitive
to denaturation reactions. Denaturation is the unfold-
ing of the enzyme tertiary structure to a disordered
polypeptide, which may lead to an irreversible loss of
activity or inactivation (Iyer and Ananthanarayan,
2008). Exposure of phytases to heat, added moisture,
and mechanical pressure are factors that lead to dena-
turation of the enzyme (Ward, 2002). For animal feeds,
the largest potential for denaturation in feed processing
is pelleting, where temperatures may reach 60 to 90°C.
Aspergillus niger phytase, which was the first com-
mercially available phytase, yielded the most activity
compared with other microorganisms tested (Shieh and
Ware, 1968); however, it was stable only to 63°C (Ullah
and Gibson, 1987). Potentially greater losses in phytase
activity may be incurred if expanders or extruders are
used to produce the diets because feed is subjected to
temperatures as high as 115 to 140°C (Fancher, 1996).
Therefore, optimizing the thermostability of exogenous
phytases, especially during feed processing, has received
much attention in the past decade and has contributed
significantly to the expanded use of phytases in swine
and poultry diets.
It is common practice for phytase manufacturers to
provide overages in phytase activity to account for po-
tential losses during feed processing treatments and
storage. However, data are limited regarding the stor-
age stability (defined as percentage of initial phytase
activity) of commercial phytases, except for those re-
ported by manufacturers in product registrations (Eu-
ropean Food Safety Authority, 2006, 2008, 2009). Al-
though temperatures and conditions from manufacture,
transport, and storage of phytases may not approxi-
mate conditions during feed processing, enough varia-
tion occurs in storage conditions and time among phy-
tase users to expect further losses in phytase activity.
Most nutritionists do not measure phytase activity at
the time of use; therefore, understanding that the sta-
bility of different commercial phytases during storage is
affected by temperature and time is important.
The results of this study demonstrated that when
phytase is stored at room temperature (23°C) or lower,
the pure product retained most (approximately 85%) of
its activity up to 60 d of storage regardless of the phy-
tase source or coating. However, phytase source influ-
enced stability when storing the product for more than
60 d at 23°C or lower, with OptiPhos and Phyzyme
retaining more activity than Ronozyme. In our study,
coated Phyzyme XP 5000 G and Phyzyme XP 10000
TPT retained 90.9 and 86.3% of their initial activity,
respectively, when stored at 23°C for 180 d; these reten-
tion rates are similar to those reported to the European
Food Safety Authority (2006, 2008). In these reports,
the product had 87 and 80% of its initial activity after
365 d of storage at 20°C. However, our results did not
confirm the retention rates reported for uncoated Rono-
zyme P-M (European Food Safety Authority, 2009).
It was reported that after 180 d, uncoated Ronozyme
P-M retained 99 and 90% of its initial phytase activity
when stored at 10 and 25°C, respectively, which was
Figure 6. Residual phytase activity (% of initial) for Ronozyme
P-CT (coated; DSM Nutritional Products, Basel, Switzerland) and
Ronozyme P-M (uncoated; DSM Nutritional Products) as affected by
form of storage [as pure product (PUR), in a vitamin premix (VIT),
or in a vitamin and trace mineral premix (VTM)], storage temperature
[room temperature (23°C) and controlled environment chamber (37°C
and 75% humidity)], and time (30 to 360 d). Each data point (least
squares means ± 3.75) is the mean of 3 observations.
4269Storage stability of phytase
9. greater than our observations (58.7% for 120 d at 5°C
and 60.6% after 180 d at 23°C).
Storing phytase in ambient temperatures greater than
23°C with high humidity proved detrimental to the sta-
bility of the pure product. More important, phytase
source affected retention rates with increasing storage
time, with the greatest rates recovered from OptiPhos,
followed by Phyzyme and, finally, by Ronozyme. This
ranking of the 3 phytase sources persisted throughout
the study. The difference in retained phytase activities
was extremely large between OptiPhos and Ronozyme
(91.5 vs. 52.6% after 30 d, 45.8 vs. 8.9% after 180 d). In
the European Food Safety Authority (2009) report, un-
coated Ronozyme P-M kept at 40°C and 60% relative
humidity retained only 50% of its initial phytase activi-
ty after 30 d, which is similar to the rate retained in our
study. The stability limit of Escherichia coli phytases,
such as OptiPhos and Phyzyme, is 60°C (Golovan et
al., 2000), whereas the stability limit for Peniophora
lycii phytases is 80°C (Phillippy, 2002). Both tempera-
tures are higher than the heat treatment used in this
study; however, one major difference is that these ther-
mal stability rates were determined by incubating the
enzyme at low pH for a short duration (approximately
30 min), whereas the enzyme was subjected to lower
but sustained heat for a longer duration (up to 180
d) in this study. Another factor may be the high hu-
midity (75%) in the chambers in our study. Yang et
al. (2007) evaluated the effects of increasing ambient
humidity (from 53 to 90%) on the stability of com-
mercial phytases stored at high ambient temperatures
(40°C for 70 d) and observed that phytase activity de-
creased significantly with increasing ambient humidity.
This suggests that regardless of the phytase source, the
environmental conditions set in the current study were
sufficient to denature the enzyme and reduce activity.
These conditions did not attempt to mimic real condi-
tions during transport or storage of the product, when
temperatures and humidities may be more variable, but
the results demonstrate the importance of maintaining
good conditions (e.g., 23°C or lower and low ambient
humidity) during storage to achieve greater stability
from phytase products.
Fat or carbohydrate coating of phytases is one widely
adopted technology used to improve phytase thermo-
stability during feed processing. Numerous studies on
the effects of coating have focused more on retention
of phytase activity after feed processing (Eeckhout,
2002; Sorbara et al., 2007) or on effects on subsequent
growth performance (Emiola et al., 2007; Timmons et
al., 2008). No study has evaluated the effects of coat-
ing on the storage stability of phytase products. Over-
all, coated pure products had greater phytase activities
than uncoated pure products when exposed to 37°C
and increased storage time, but this differed between
phytase sources. Coating was beneficial for Ronozyme
and OptiPhos from d 90 onward, but not for Phyzyme,
for which the uncoated product retained more activity
than the coated product throughout the study. This
suggests that the type of coating may differ between
phytase manufacturers and that some coated phytase
products may provide better protection during storage
than others.
Phytases in Premixes
For most of the commercial phytase products tested,
retained phytase activities were more than 85, 73, and
60% of their initial activity up to 180 d when stored
as pure products, vitamin premixes, or VTM premix-
es, respectively, and when storage temperatures were
23°C. The exceptions were uncoated Ronozyme P-M
for the pure phytase products, uncoated Phyzyme XP
5000 G for the vitamin premixes, and uncoated Opti-
Phos 2000-M for the VTM premixes. Previously, the
stability of coated Phyzyme XP 10000 TPT was tested
in a VTM premix that was stored at 20°C for 180 d
(European Food Safety Authority, 2008). The prod-
uct retained 76% if its initial phytase activity, which is
less than the 92% observed in our study. The stability
of uncoated Ronozyme P-M was also evaluated previ-
ously when mixed with VTM premixes and stored at
25°C (European Food Safety Authority, 2009). After 90
d, 89% of the initial activity was retained, which was
greater than the 77% observed in this study. In general,
greater retention was observed with increasing storage
time when phytases were stored as pure products than
when they were mixed into either of the premixes. This
suggests that storing phytases in pure forms may have
advantages in retaining the original phytase activity
compared with including it in premixes when stored at
room (23°C) temperature or lower.
When phytase was mixed into vitamin or VTM pre-
mixes and exposed to heat treatment (37°C), coated
phytases retained greater activities than uncoated phy-
tases, especially when stored for more than 90 d. How-
ever, some differences surfaced among phytase sources
in that coating had the greatest benefits for OptiPhos.
Results also showed that uncoated phytases had very
poor stability when stored as premixes for as little as
30 d. The loss of phytase activity was greater when
phytase was mixed with VTM premixes than when it
was mixed with vitamin premixes, suggesting that high
heat and humidity as well as potential interactions with
some components of the premixes increased the rate of
phytase denaturation. Previous work has shown that
mixing inorganic trace minerals with vitamins leads to
significant losses in vitamin activity, which is thought
to be due to the presence of ionic charges in mineral
salts that can act as oxidizing agents (Shurson et al.,
1996). The objective of this study was not to identify
specific vitamins or trace minerals that may have con-
tributed to greater losses in phytase activity, but re-
sults indicate that that coating mitigated the negative
effects of heat and humidity for Ronozyme and Opti-
Phos, but not Phyzyme. This study also demonstrated
the differing abilities of coating technologies to protect
phytases against not only environmental degradation
4270 Sulabo et al.
10. but also the negative effects of certain components in
vitamin and VTM premixes.
The stability of commercially available phytases dur-
ing storage is affected by numerous factors, such as
storage time, temperature, product form, coating, and
source. Pure phytase products stored at 23°C or lower
were the most stable. In premixes, longer storage times
and higher temperatures reduced phytase activity, but
coating mitigated some of these negative effects.
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4271Storage stability of phytase