This document discusses a study on the effects of civet coffee isolate concentration and fermentation time on the protein profiles of Robusta coffee. Robusta coffee beans were fermented in vitro using different concentrations of civet coffee microbial isolates from feces and incubation times ranging from 24 to 60 hours. The protein concentration and molecular weight profiles of the fermented coffee beans were then analyzed. The results showed that the protein concentration decreased with longer fermentation times and higher isolate concentrations, with the lowest concentration at 50.95 μg/mL after 60 hours of fermentation with the highest isolate concentration. The number of protein bands also increased with higher isolate concentrations.
Microwave assisted extraction was used to obtain water soluble extracts from spent coffee residues that were then added to cookies to increase their antioxidant properties. Two sequential MAE extractions obtained 19% and 21% soluble material from the coffee residues. The extracts contained chlorogenic acids, particularly 3-, 4-, and 5-O-caffeoylquinic acids which made up 67-71% of the total chlorogenic acids extracted. Adding these extracts to cookies in amounts up to 3% increased the antioxidant activity of the cookies by up to 62% without negatively affecting taste, maintaining antioxidant properties for over 55 days of storage. Thus, MAE is an efficient technique for extracting bioactive compounds from coffee residues that can be used as functional food add
Production of Phycocyanin. and efficient methods for their extraction.Niyamat Panjesha
This document presents a project report on cultivating Spirulina spp. using sugar industry effluent to produce phycocyanin. The objectives were to characterize the effluent, study Spirulina growth and phycocyanin production. The effluent was analyzed and found suitable for growth. Spirulina was cultivated and growth was monitored over 5 days. Phycocyanin was extracted using different methods, with homogenization yielding the highest amount. Cultivating Spirulina in effluent can produce phycocyanin more cost effectively.
This document discusses a study on increasing the ratio of rice to barley that can be used in brewing beer. It found that rice naturally contains lower levels of soluble protein compared to barley, limiting how much rice can be used. The study tested adding the protease enzyme Neutrase and modifying mashing methods to increase soluble nitrogen levels in wort made from rice. It determined that mashing rice for 60 minutes and adding 400 μl of Neutrase per 50g of rice was optimal. Activating another enzyme also increased solubilized protein from rice. Various rice-barley mixtures were then mashed and fermented, finding worts with sufficient nitrogen levels could contain up to 80% rice. Therefore, enzyme addition and mashing
This study investigated enzymatic depolymerisation of beta-glucan from oat bran to modify its molecular weight and viscosity properties. Two oat bran concentrates with different beta-glucan contents were treated with commercial enzyme mixtures and a purified endoglucanase at both high and low water contents. Treatment parameters like enzyme type, dosage, and reaction time significantly affected beta-glucan hydrolysis. The suitable range of beta-glucan molecular weight was obtained by treating oat bran concentrate at 50% moisture content for over 3 hours with a specific commercial enzyme preparation, resulting in a peak average molecular weight of 47 kDa for the beta-glucan. This low molecular weight beta-gluc
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
MEASUREMENT OF CAFFEINE IN COFFEE BEANS USING UV/VIS SPECTROPHOTOMETRY.docxukponganthony03
This document outlines a seminar presentation on measuring caffeine content in coffee beans using UV/VIS spectrophotometry. The presentation aims to develop a reliable method for caffeine quantification in coffee beans using this analytical technique. It will involve preparing caffeine standards of known concentrations, extracting caffeine from coffee bean samples using a solvent, measuring absorbance of the solutions, and constructing a calibration curve to correlate absorbance with caffeine concentration. The expected results are a strong linear relationship between concentration and absorbance, allowing accurate determination of caffeine levels in coffee beans.
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
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.
Microwave assisted extraction was used to obtain water soluble extracts from spent coffee residues that were then added to cookies to increase their antioxidant properties. Two sequential MAE extractions obtained 19% and 21% soluble material from the coffee residues. The extracts contained chlorogenic acids, particularly 3-, 4-, and 5-O-caffeoylquinic acids which made up 67-71% of the total chlorogenic acids extracted. Adding these extracts to cookies in amounts up to 3% increased the antioxidant activity of the cookies by up to 62% without negatively affecting taste, maintaining antioxidant properties for over 55 days of storage. Thus, MAE is an efficient technique for extracting bioactive compounds from coffee residues that can be used as functional food add
Production of Phycocyanin. and efficient methods for their extraction.Niyamat Panjesha
This document presents a project report on cultivating Spirulina spp. using sugar industry effluent to produce phycocyanin. The objectives were to characterize the effluent, study Spirulina growth and phycocyanin production. The effluent was analyzed and found suitable for growth. Spirulina was cultivated and growth was monitored over 5 days. Phycocyanin was extracted using different methods, with homogenization yielding the highest amount. Cultivating Spirulina in effluent can produce phycocyanin more cost effectively.
This document discusses a study on increasing the ratio of rice to barley that can be used in brewing beer. It found that rice naturally contains lower levels of soluble protein compared to barley, limiting how much rice can be used. The study tested adding the protease enzyme Neutrase and modifying mashing methods to increase soluble nitrogen levels in wort made from rice. It determined that mashing rice for 60 minutes and adding 400 μl of Neutrase per 50g of rice was optimal. Activating another enzyme also increased solubilized protein from rice. Various rice-barley mixtures were then mashed and fermented, finding worts with sufficient nitrogen levels could contain up to 80% rice. Therefore, enzyme addition and mashing
This study investigated enzymatic depolymerisation of beta-glucan from oat bran to modify its molecular weight and viscosity properties. Two oat bran concentrates with different beta-glucan contents were treated with commercial enzyme mixtures and a purified endoglucanase at both high and low water contents. Treatment parameters like enzyme type, dosage, and reaction time significantly affected beta-glucan hydrolysis. The suitable range of beta-glucan molecular weight was obtained by treating oat bran concentrate at 50% moisture content for over 3 hours with a specific commercial enzyme preparation, resulting in a peak average molecular weight of 47 kDa for the beta-glucan. This low molecular weight beta-gluc
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
MEASUREMENT OF CAFFEINE IN COFFEE BEANS USING UV/VIS SPECTROPHOTOMETRY.docxukponganthony03
This document outlines a seminar presentation on measuring caffeine content in coffee beans using UV/VIS spectrophotometry. The presentation aims to develop a reliable method for caffeine quantification in coffee beans using this analytical technique. It will involve preparing caffeine standards of known concentrations, extracting caffeine from coffee bean samples using a solvent, measuring absorbance of the solutions, and constructing a calibration curve to correlate absorbance with caffeine concentration. The expected results are a strong linear relationship between concentration and absorbance, allowing accurate determination of caffeine levels in coffee beans.
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
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.
Proximate and Microbial Profile of Couscous Yoghurt Produced from Soya MilkIJEAB
The document summarizes a study that investigated the effect of milk type (cow milk, soya milk, or a 50:50 mixture) and mixing ratio on the proximate composition and microbial profile of couscous yoghurt. Yoghurts were produced from the three milk types then mixed with couscous at ratios of 90:10, 80:20, and 70:30. The experiment tested 9 treatments total. Proximate compositions and microbial counts were analyzed. Results showed significant differences in composition and counts based on milk type and ratio. The cow-soya milk mixture at a 70:30 ratio had the highest protein and nutrient content. All samples had microbial levels within acceptable standards.
BIO DECAFFEINATION-A STUDY ON THE EFFECTS OF BREVIBACTERIUM ON DIFFERENT SAMP...EDITOR IJCRCPS
HPLC analysis of caffeine was performed in SHIMADZU LC 20 – AD system, and the caffeine compounds were separated on a
C18 column under isocratic conditions with 40% methanol in water at a flow rate of 1.0 ml/min. Compounds eluting from the
column were detected and the peak areas were compared with those obtained with standards of known concentration. The HPLC
analysis of caffeine degradation by Brevibacterium is done by injecting the sample volume of about 20μl HPLC analysis is done
for the sample at different incubation periods with standard caffeine concentration (Known). The sample is analyzed for every
twelve hours of incubation and peak values are obtained. Caffeine concentration is an important parameter to be checked as
excessive consumption of caffeine leads to many health hazards.
Keywords: , Biodecaffeination, Brevibacterium, HPLC.
Comparative Study of Production of Single Cell Protein from Different Agricul...ijtsrd
Single cell protein SCP also referred as microbial protein is defined as protein derived from cells of microorganisms such as yeast, fungi, algae, and bacteria, which are grown on various carbon sources for synthesis. The dried cells of microorganisms or the whole organism is harvested and consumed. In this work SCP was produced from different agricultural waste substrates like food and vegetable waste, rice husk, pulses husk, bagasse and wheat straw using Aspergillus niger. These substrates not only act as nutritive supplement but also ensure good waste management. Also, carbohydrate content of each sample was determined. For maximizing the yield of SCP, some factors were optimized. Various buffers were used like phosphate buffer, carbonate bicarbonate buffer and 0.1N NaOH. The sample that shows the best result for SCP was identified to be MCD and fruit and vegetable waste in 50 50 ratio and rice husk. In the future SCP could be produced to not only be used to produce protein but multiple products rich in carbohydrate, vitamins, lipids and other amino acids. Also yield could be increased by genetically modifying SCP organisms. Abhishikta Dasgupta | Jasmine Chughasrani "Comparative Study of Production of Single Cell Protein from Different Agricultural Waste Substrates using Aspergillus Niger" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-2 , February 2021, URL: https://www.ijtsrd.com/papers/ijtsrd38339.pdf Paper Url: https://www.ijtsrd.com/home-science/food-biotechnology/38339/comparative-study-of-production-of-single-cell-protein-from-different-agricultural-waste-substrates-using-aspergillus-niger/abhishikta-dasgupta
JBEI Research Highlights - October 2021SaraHarmon4
This document summarizes a study that evaluated the potential for nutrient recovery from wet organic waste processing facilities in California to offset synthetic fertilizer demand. The study found that recovering nitrogen and phosphorus from organic waste streams through anaerobic digestion and separation techniques could meet 11% of the state's nitrogen and 29% of phosphorus fertilizer needs. Recovered nutrients would be in the form of liquid fertilizer, struvite, and compost. The approach provides a foundation for analyzing national-level nutrient flows and recovery potentials from bioenergy production.
The study analyzed the effect of common poultry medicines on ethanol fermentation using Saccharomyces cerevisiae. Various concentrations of Avatec, Cygro, Robenz, and Monesine were tested in batch reactions at 33°C and 36°C. Results showed that higher concentrations of Robenz and Avatec decreased sucrose concentration over time, suggesting reduced bacterial contaminants and increased yeast growth. Specifically, Robenz 100 ppm/L and Avatec 1000 ppm/L at 33°C, and Avatec 100 ppm/L at 36°C minimized bacterial growth. However, very high medicine concentrations also reduced yeast. Further analysis is needed to differentiate sucrose consumption by yeast versus bacteria to
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.
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
The annual Research Poster Session at the conference features cutting-edge food safety research related to fresh and fresh-cut produce from researchers around the world. Posters will be on display during the conference and researchers will be available at their posters on June 21 from 2-4pm to discuss their research. The document then provides summaries of 4 research posters that will be presented on topics including the antimicrobial effects of haskap berry extracts on foodborne pathogens, using whole genome sequencing and genetic analysis to map contamination sources in produce packing facilities, developing alternative seed disinfection methods for sprouted vegetables, and developing methods to encapsulate ethylene to control fruit ripening.
Effects of Fermentation of Cashew Kernel on the Nutrient Value of Cassava Sem...Agriculture Journal IJOEAR
— Protein-energy malnutrition in children is a public health problem. This nutrition problem is attributed to inappropriate complementary feeding. Indeed, the cost of high-quality food supplements is high and traditional food supplements have a low nutritional quality related to the presence of antinutritional factors. The objective of this study is to determine acceptability and antinutritional factors in attiéké / cashew kernel composite flours. The cashew kernel flour is produced after various technological treatments to obtain two types of flour (unfermented flour and fermented flour). Physico-chemical and sensory analyzes are performed. The results showed that fermentation has an influence on the parameters studied. The protein contents of the unfermented formulations range from 7.53% to 10.62% while those of the fermented formulations range from 8.23% to 11.53%. Both formulations contain antinutritional factors.
Evaluation of the Glucuronic Acid Production and Other Biological Activities...IJMER
The document evaluates the production of glucuronic acid and other biological activities of fermented sweetened black tea fermented with Kombucha culture alone or in combination with different Lactobacillus strains isolated from Kefir grains. Key findings include:
1) Lactobacillus casei increased glucuronic acid production in fermented tea by 39.6% compared to Kombucha culture alone.
2) Lactobacillus plantarum enhanced the antibacterial and antioxidant activities of fermented tea to a greater level than normal fermented tea or mixes with other Lactobacillus strains.
3) The study suggests certain Lactobacillus strains from Kefir grains
This document describes a study that used the seed powder of Strychnos potatorum (clearing nut) to harvest the microalga Chlorella vulgaris through a process called bioflocculation. The researchers optimized bioflocculation parameters like bioflocculant concentration, temperature, agitation speed, and incubation time using Response Surface Methodology. They found that 100 mg/L bioflocculant concentration, 35°C temperature, 150 rpm agitation, and 30 minutes incubation time resulted in maximum bioflocculation efficiency of 99.68%. A cell viability test showed cells remained intact after bioflocculation but were destroyed using a chemical flocculant, indicating bioflocculation's advantages. The
This document discusses a study that characterized Ghanaian cocoa bean fermentation using spectroscopic and chromatographic methods. Researchers used colorimetry, fluorescence spectroscopy, near-infrared spectroscopy, and gas chromatography-mass spectrometry to examine cocoa beans sampled at different stages of fermentation. The degree of fermentation could be described well by the spectroscopic methods. Certain aroma compounds like 2-phenylethyl acetate increased during fermentation while others like diacetyl decreased. The study demonstrates the potential of using spectroscopy to objectively determine cocoa bean quality and degree of fermentation.
54.Isolation and purification of cellulase from Aspergillus terreusAnnadurai B
This document describes the isolation and purification of cellulase enzymes from the fungus Aspergillus terreus. The intracellular cellulase was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and gel filtration chromatography. This purification scheme achieved a 270-fold purification with a 22.11% yield. Tests including PAGE, SDS-PAGE, immunodiffusion, and isoelectric focusing confirmed the homogeneity of the purified enzyme. The purified cellulase showed optimal activity between pH 4-7 and temperatures of 40-50°C.
The document describes optimization of the fermentation medium for production of biomass and nattokinase by Bacillus subtilis natto. Initial tests confirmed the bacterium isolated from Vietnamese natto food was Bacillus subtilis natto. Six factors in the fermentation medium were screened using Plackett-Burman design, identifying soybean peptone and CaCl2 as significant for biomass production. Response surface methodology was used to optimize the medium for highest dried cell weight of 3.033 g/L. This optimized medium increased nattokinase yield by over 30% to 31.06 FU/mL compared to the initial medium.
This document discusses upgrading exoglucanase production by Trichoderma reesei NRC 210 through fermentation process optimization. Batch fermentations were performed to study the effects of pH and agitation speed on enzyme production. Results showed that controlling pH from 4-5 and increasing agitation speed to 350 rpm increased exoglucanase activity by about 15-fold and 1.8-fold, respectively. Kinetic models including the Leudeking-Piret and Monod models were applied and showed good fit to experimental data, indicating growth-associated exoglucanase production and substrate consumption patterns over fermentation time. Process optimization led to improved exoglucanase yields useful for industrial applications.
International Journal of Engineering Research and DevelopmentIJERD Editor
Electrical, Electronics and Computer Engineering,
Information Engineering and Technology,
Mechanical, Industrial and Manufacturing Engineering,
Automation and Mechatronics Engineering,
Material and Chemical Engineering,
Civil and Architecture Engineering,
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Marine and Agriculture engineering,
Aerospace Engineering.
Production of banana alcohol and utilization of banana residueeSAT Journals
Abstract Aim of the study was production of alcohol from banana juice which use as complete replacement of malt in alcohol production by utilizing pure culture of Sacharomyces cerevisiae as fermenting organism. Banana juice was made from banana pulp by using pectinase enzyme. Optimization of amount of pectinase enzyme for juice production and optimization of pH of the final product were also aim of this study. Pectinase enzyme used for liquefying the pulp production was 0.0003% (w/v). The sugar percentage found in the banana juice was 18%. A sequential study has been done by consecutive pH levels of 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0 in the final product. The best product was obtained at pH 6.0 with respect to taste; pH was regulated only after the complete fermentation of the banana juice but just before the filtration process. Alcohol percentage of the product was 8% (v/v) at 28oC. Total number of colonies detected was 21 in freshly prepared alcohol and total number of colonies detected was 20 in the beer after 5 months from production. Another aim of the work was utilization of the banana residue for the production of fiber enriched cookies. High fiber enriched cookies were prepared using 5%-20% level of fiber obtained from banana residue. 7%-10 % fiber content was obtained as best parameter for cookie production and final moisture content of cookie was 3%. Keywords: banana pulp, depectinization, pectinase enzyme, Sacharomyces cerevisiae, alcohol, banana fiber, cookies.
CULTIVATION OF OSCILLATORIA SP IN DAIRY WASTE WATER IN TWO STAGE PHOTO BIOREA...civej
This paper presents an integrated approach to cultivate microalgae in dairy wastewater and to
investigate the capability of the organism for biodiesel production. The present study was carried out
using tolerant strains of microalgae collected from dairy effluent treatment plant, Kochi. Selected blue
green algal strains were mass cultured in the laboratory and acclimatized using different concentrations
of synthetic effluent. Blue green algal filaments were immobilized inside the primary and secondary
photobioreactors. The experiment was conducted in two stages including batch and continuous
treatment. The stage 1 of the experiment was designed for the reduction of physical impurities and the
nutrients. Stage 2 was designed mainly for the cultivation of blue green algae in dairy waste water by
utilizing the extra nutrients . Reduction of 94 -99.5% in phosphate was observed after 48 h of treatment
in the primary and secondary photobioreactors. The level of phosphate, total hardness, ammoniacal
nitrogen in the MSE was reduced by 97%,93 %, 81% respectively. BOD was reduced to 370mg L-1 from
1500 mg L-1 after 48 hrs of treatment in the primary reactor. COD was reduced to 85 mg L -1 from an
initial value of 1500 mg L -1 from medium strength effluent (MSE) and 90-95 % removal of COD was
also obtained from high strength effluent(HSE) during the study period. Biomass developed within the
reactor was harvested at every 15 days intervals from the secondary reactor and analyzed for lipids and
fattyacids. Presence of C14:0, C16:0,C18:0, C18:1 and C18:2 fatty acids strongly supports its abilility for
biodiesel production.
The results show that UAE extraction provides higher yields of caffeine (6-8%) and 5-CQA (0.8-0.9%) from spent coffee grounds than magnetic stirring, but the benefits are time dependent. Robusta coffee grounds contain more caffeine and 5-CQA than Arabica grounds and have greater antioxidant activity. While extraction yields are low, the large quantities of spent coffee grounds produced annually represent a potential source of natural antioxidants and caffeine if cost-effective extraction methods can be developed.
Proximate and Microbial Profile of Couscous Yoghurt Produced from Soya MilkIJEAB
The document summarizes a study that investigated the effect of milk type (cow milk, soya milk, or a 50:50 mixture) and mixing ratio on the proximate composition and microbial profile of couscous yoghurt. Yoghurts were produced from the three milk types then mixed with couscous at ratios of 90:10, 80:20, and 70:30. The experiment tested 9 treatments total. Proximate compositions and microbial counts were analyzed. Results showed significant differences in composition and counts based on milk type and ratio. The cow-soya milk mixture at a 70:30 ratio had the highest protein and nutrient content. All samples had microbial levels within acceptable standards.
BIO DECAFFEINATION-A STUDY ON THE EFFECTS OF BREVIBACTERIUM ON DIFFERENT SAMP...EDITOR IJCRCPS
HPLC analysis of caffeine was performed in SHIMADZU LC 20 – AD system, and the caffeine compounds were separated on a
C18 column under isocratic conditions with 40% methanol in water at a flow rate of 1.0 ml/min. Compounds eluting from the
column were detected and the peak areas were compared with those obtained with standards of known concentration. The HPLC
analysis of caffeine degradation by Brevibacterium is done by injecting the sample volume of about 20μl HPLC analysis is done
for the sample at different incubation periods with standard caffeine concentration (Known). The sample is analyzed for every
twelve hours of incubation and peak values are obtained. Caffeine concentration is an important parameter to be checked as
excessive consumption of caffeine leads to many health hazards.
Keywords: , Biodecaffeination, Brevibacterium, HPLC.
Comparative Study of Production of Single Cell Protein from Different Agricul...ijtsrd
Single cell protein SCP also referred as microbial protein is defined as protein derived from cells of microorganisms such as yeast, fungi, algae, and bacteria, which are grown on various carbon sources for synthesis. The dried cells of microorganisms or the whole organism is harvested and consumed. In this work SCP was produced from different agricultural waste substrates like food and vegetable waste, rice husk, pulses husk, bagasse and wheat straw using Aspergillus niger. These substrates not only act as nutritive supplement but also ensure good waste management. Also, carbohydrate content of each sample was determined. For maximizing the yield of SCP, some factors were optimized. Various buffers were used like phosphate buffer, carbonate bicarbonate buffer and 0.1N NaOH. The sample that shows the best result for SCP was identified to be MCD and fruit and vegetable waste in 50 50 ratio and rice husk. In the future SCP could be produced to not only be used to produce protein but multiple products rich in carbohydrate, vitamins, lipids and other amino acids. Also yield could be increased by genetically modifying SCP organisms. Abhishikta Dasgupta | Jasmine Chughasrani "Comparative Study of Production of Single Cell Protein from Different Agricultural Waste Substrates using Aspergillus Niger" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-5 | Issue-2 , February 2021, URL: https://www.ijtsrd.com/papers/ijtsrd38339.pdf Paper Url: https://www.ijtsrd.com/home-science/food-biotechnology/38339/comparative-study-of-production-of-single-cell-protein-from-different-agricultural-waste-substrates-using-aspergillus-niger/abhishikta-dasgupta
JBEI Research Highlights - October 2021SaraHarmon4
This document summarizes a study that evaluated the potential for nutrient recovery from wet organic waste processing facilities in California to offset synthetic fertilizer demand. The study found that recovering nitrogen and phosphorus from organic waste streams through anaerobic digestion and separation techniques could meet 11% of the state's nitrogen and 29% of phosphorus fertilizer needs. Recovered nutrients would be in the form of liquid fertilizer, struvite, and compost. The approach provides a foundation for analyzing national-level nutrient flows and recovery potentials from bioenergy production.
The study analyzed the effect of common poultry medicines on ethanol fermentation using Saccharomyces cerevisiae. Various concentrations of Avatec, Cygro, Robenz, and Monesine were tested in batch reactions at 33°C and 36°C. Results showed that higher concentrations of Robenz and Avatec decreased sucrose concentration over time, suggesting reduced bacterial contaminants and increased yeast growth. Specifically, Robenz 100 ppm/L and Avatec 1000 ppm/L at 33°C, and Avatec 100 ppm/L at 36°C minimized bacterial growth. However, very high medicine concentrations also reduced yeast. Further analysis is needed to differentiate sucrose consumption by yeast versus bacteria to
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.
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
The annual Research Poster Session at the conference features cutting-edge food safety research related to fresh and fresh-cut produce from researchers around the world. Posters will be on display during the conference and researchers will be available at their posters on June 21 from 2-4pm to discuss their research. The document then provides summaries of 4 research posters that will be presented on topics including the antimicrobial effects of haskap berry extracts on foodborne pathogens, using whole genome sequencing and genetic analysis to map contamination sources in produce packing facilities, developing alternative seed disinfection methods for sprouted vegetables, and developing methods to encapsulate ethylene to control fruit ripening.
Effects of Fermentation of Cashew Kernel on the Nutrient Value of Cassava Sem...Agriculture Journal IJOEAR
— Protein-energy malnutrition in children is a public health problem. This nutrition problem is attributed to inappropriate complementary feeding. Indeed, the cost of high-quality food supplements is high and traditional food supplements have a low nutritional quality related to the presence of antinutritional factors. The objective of this study is to determine acceptability and antinutritional factors in attiéké / cashew kernel composite flours. The cashew kernel flour is produced after various technological treatments to obtain two types of flour (unfermented flour and fermented flour). Physico-chemical and sensory analyzes are performed. The results showed that fermentation has an influence on the parameters studied. The protein contents of the unfermented formulations range from 7.53% to 10.62% while those of the fermented formulations range from 8.23% to 11.53%. Both formulations contain antinutritional factors.
Evaluation of the Glucuronic Acid Production and Other Biological Activities...IJMER
The document evaluates the production of glucuronic acid and other biological activities of fermented sweetened black tea fermented with Kombucha culture alone or in combination with different Lactobacillus strains isolated from Kefir grains. Key findings include:
1) Lactobacillus casei increased glucuronic acid production in fermented tea by 39.6% compared to Kombucha culture alone.
2) Lactobacillus plantarum enhanced the antibacterial and antioxidant activities of fermented tea to a greater level than normal fermented tea or mixes with other Lactobacillus strains.
3) The study suggests certain Lactobacillus strains from Kefir grains
This document describes a study that used the seed powder of Strychnos potatorum (clearing nut) to harvest the microalga Chlorella vulgaris through a process called bioflocculation. The researchers optimized bioflocculation parameters like bioflocculant concentration, temperature, agitation speed, and incubation time using Response Surface Methodology. They found that 100 mg/L bioflocculant concentration, 35°C temperature, 150 rpm agitation, and 30 minutes incubation time resulted in maximum bioflocculation efficiency of 99.68%. A cell viability test showed cells remained intact after bioflocculation but were destroyed using a chemical flocculant, indicating bioflocculation's advantages. The
This document discusses a study that characterized Ghanaian cocoa bean fermentation using spectroscopic and chromatographic methods. Researchers used colorimetry, fluorescence spectroscopy, near-infrared spectroscopy, and gas chromatography-mass spectrometry to examine cocoa beans sampled at different stages of fermentation. The degree of fermentation could be described well by the spectroscopic methods. Certain aroma compounds like 2-phenylethyl acetate increased during fermentation while others like diacetyl decreased. The study demonstrates the potential of using spectroscopy to objectively determine cocoa bean quality and degree of fermentation.
54.Isolation and purification of cellulase from Aspergillus terreusAnnadurai B
This document describes the isolation and purification of cellulase enzymes from the fungus Aspergillus terreus. The intracellular cellulase was purified using ammonium sulfate precipitation, DEAE-cellulose chromatography, and gel filtration chromatography. This purification scheme achieved a 270-fold purification with a 22.11% yield. Tests including PAGE, SDS-PAGE, immunodiffusion, and isoelectric focusing confirmed the homogeneity of the purified enzyme. The purified cellulase showed optimal activity between pH 4-7 and temperatures of 40-50°C.
The document describes optimization of the fermentation medium for production of biomass and nattokinase by Bacillus subtilis natto. Initial tests confirmed the bacterium isolated from Vietnamese natto food was Bacillus subtilis natto. Six factors in the fermentation medium were screened using Plackett-Burman design, identifying soybean peptone and CaCl2 as significant for biomass production. Response surface methodology was used to optimize the medium for highest dried cell weight of 3.033 g/L. This optimized medium increased nattokinase yield by over 30% to 31.06 FU/mL compared to the initial medium.
This document discusses upgrading exoglucanase production by Trichoderma reesei NRC 210 through fermentation process optimization. Batch fermentations were performed to study the effects of pH and agitation speed on enzyme production. Results showed that controlling pH from 4-5 and increasing agitation speed to 350 rpm increased exoglucanase activity by about 15-fold and 1.8-fold, respectively. Kinetic models including the Leudeking-Piret and Monod models were applied and showed good fit to experimental data, indicating growth-associated exoglucanase production and substrate consumption patterns over fermentation time. Process optimization led to improved exoglucanase yields useful for industrial applications.
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Petroleum and Mining Engineering,
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Aerospace Engineering.
Production of banana alcohol and utilization of banana residueeSAT Journals
Abstract Aim of the study was production of alcohol from banana juice which use as complete replacement of malt in alcohol production by utilizing pure culture of Sacharomyces cerevisiae as fermenting organism. Banana juice was made from banana pulp by using pectinase enzyme. Optimization of amount of pectinase enzyme for juice production and optimization of pH of the final product were also aim of this study. Pectinase enzyme used for liquefying the pulp production was 0.0003% (w/v). The sugar percentage found in the banana juice was 18%. A sequential study has been done by consecutive pH levels of 4.5, 5.0, 5.5, 6.0, 6.5, and 7.0 in the final product. The best product was obtained at pH 6.0 with respect to taste; pH was regulated only after the complete fermentation of the banana juice but just before the filtration process. Alcohol percentage of the product was 8% (v/v) at 28oC. Total number of colonies detected was 21 in freshly prepared alcohol and total number of colonies detected was 20 in the beer after 5 months from production. Another aim of the work was utilization of the banana residue for the production of fiber enriched cookies. High fiber enriched cookies were prepared using 5%-20% level of fiber obtained from banana residue. 7%-10 % fiber content was obtained as best parameter for cookie production and final moisture content of cookie was 3%. Keywords: banana pulp, depectinization, pectinase enzyme, Sacharomyces cerevisiae, alcohol, banana fiber, cookies.
CULTIVATION OF OSCILLATORIA SP IN DAIRY WASTE WATER IN TWO STAGE PHOTO BIOREA...civej
This paper presents an integrated approach to cultivate microalgae in dairy wastewater and to
investigate the capability of the organism for biodiesel production. The present study was carried out
using tolerant strains of microalgae collected from dairy effluent treatment plant, Kochi. Selected blue
green algal strains were mass cultured in the laboratory and acclimatized using different concentrations
of synthetic effluent. Blue green algal filaments were immobilized inside the primary and secondary
photobioreactors. The experiment was conducted in two stages including batch and continuous
treatment. The stage 1 of the experiment was designed for the reduction of physical impurities and the
nutrients. Stage 2 was designed mainly for the cultivation of blue green algae in dairy waste water by
utilizing the extra nutrients . Reduction of 94 -99.5% in phosphate was observed after 48 h of treatment
in the primary and secondary photobioreactors. The level of phosphate, total hardness, ammoniacal
nitrogen in the MSE was reduced by 97%,93 %, 81% respectively. BOD was reduced to 370mg L-1 from
1500 mg L-1 after 48 hrs of treatment in the primary reactor. COD was reduced to 85 mg L -1 from an
initial value of 1500 mg L -1 from medium strength effluent (MSE) and 90-95 % removal of COD was
also obtained from high strength effluent(HSE) during the study period. Biomass developed within the
reactor was harvested at every 15 days intervals from the secondary reactor and analyzed for lipids and
fattyacids. Presence of C14:0, C16:0,C18:0, C18:1 and C18:2 fatty acids strongly supports its abilility for
biodiesel production.
The results show that UAE extraction provides higher yields of caffeine (6-8%) and 5-CQA (0.8-0.9%) from spent coffee grounds than magnetic stirring, but the benefits are time dependent. Robusta coffee grounds contain more caffeine and 5-CQA than Arabica grounds and have greater antioxidant activity. While extraction yields are low, the large quantities of spent coffee grounds produced annually represent a potential source of natural antioxidants and caffeine if cost-effective extraction methods can be developed.
This document provides an overview of the production process for bread. It discusses the key steps including bread fermentation using yeast, various bread formulations from around the world, common types of flour and yeast used, and the standard bread making procedure which involves steps such as mixing, fermentation, proofing, baking, and cooling. The principles of fermentation and the roles of different yeast strains are also summarized.
The document discusses various processes used in milk production and preservation including pasteurization, sterilization, evaporation, and drying. It describes methods of pasteurization like high temperature short time and low temperature long time. It also explains milk products like butter, ghee, condensed milk, dry milk, and cheeses along with how enzymes and additives are used in their production.
Canning involves preserving foods in hermetically sealed containers using heat treatment to prevent spoilage. Common foods canned include fruits, vegetables, meats. The canning process involves selection, washing, grading, cutting, filling containers, exhausting air, sealing, heat processing then cooling of packaged food. Canning destroys spoilage microbes and allows storage of food without refrigeration for years if properly processed in sterile containers.
1. Journal of Physics: Conference Series
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The effect of civet coffee isolate and time fermentation on Robusta
coffee protein profiles
To cite this article: M Uliyandari et al 2021 J. Phys.: Conf. Ser. 1731 012019
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Mathematics and Science Education International Seminar (MASEIS) 2019
Journal of Physics: Conference Series 1731(2021) 012019
IOP Publishing
doi:10.1088/1742-6596/1731/1/012019
1
The effect of civet coffee isolate and time fermentation on
Robusta coffee protein profiles
M Uliyandari1,*
, S Sumpono2
and C Muslim3
1
Program Studi Pendidikan IPA, Universitas Bengkulu, Jl. WR Supratman, Kota
Bengkulu 38122, Indonesia
2
Pascasarjana Pendidikan IPA, Universitas Bengkulu, Jl. WR Supratman, Kota
Bengkulu 38122, Indonesia
3
Fakultas Matematika dan Ilmu Pengetahuan Alam, Universitas Bengkulu, Jl. WR
Supratman, Kota Bengkulu 38122, Indonesia
*mellytauliyandar@unib.ac.id
Abstract. This research aims is to determine the effect of the isolate’s concentration of civet
coffee and fermentation time on the protein concentration profile and protein molecular weight
profile of Robusta coffee which fermented in vitro, in vivo, and without fermentation. Isolates
of civet coffee made from microbial civet feces and made in five variants concentrations
(2,91x108 CFU/mL; 2,91x107CFU/mL; 2,91x 106CFU/mL, 2,91x105 CFU/mL; 2,91x104
CFU/mL). In the fermentation process coffee Robusta beans marinated with various
concentrations of coarse isolates. The coffee beans are fermented analysed by Uv-Vis
spectrophotometer and the results obtained in vitro protein concentration of the lowest 50,95
μg/mL (fermentation time of 60 hours and isolates concentration of 2,91 x 108 mg/mL), coffee
fermentation in vivo (167, 44 μg/mL), and coffee without fermentation (217,61 μg/mL). The
protein molecular weight profile is determined based on the results of electrophoresis. The
higher the concentration of isolates civet coffee in the fermentation process, the more protein
bands appear, and vice versa.
1. Introduction
One of Indonesia's leading plantation commodities that has a high export value and to provide
substantial foreign exchange for the country is coffee. About 60% of the total national coffee
production is exported to major destination countries such as the United States, Germany, and Japan
[1].
Currently, Indonesian coffee which is exported is only limited to coffee beans, not a product that is
ready for consumption. Therefore it is necessary to research the semi-wet or dry fermentation coffee
processing technology which is more easily applied by farmers. One method that needs to be
developed is the natural fermentation process of the digestive system of mongoose animals which has
been proven to produce coffee beans with better quality and flavour [2].
Civet coffee is a type of coffee that has been processed through a short fermentation in the
digestion of civet animals (Paradoxurus Hermaphroditus). The enzymes in the civet digestive tract can
produce coffee with distinctive flavour and aroma [3]. Improved quality of civet coffee flavour is
caused by low protein content and high-fat content compared to regular coffee [4]. Coffee beans that
3. Mathematics and Science Education International Seminar (MASEIS) 2019
Journal of Physics: Conference Series 1731(2021) 012019
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doi:10.1088/1742-6596/1731/1/012019
2
are usually processed into civet coffee are Robusta coffee and Arabica coffee, where these beans
generally contain minerals, caffeine, trigonelline, fat, chlorogenic, aliphatic acids, oligosaccharides,
polysaccharides, amino acids, proteins, and human acids [5].
Bengkulu Province is one of the Robusta coffee-producing provinces in Indonesia, especially in the
Rejang Lebong district. Robusta superior grade coffee can produce coffee beans throughout the year,
more resistant to pests and leaf rust, and resistant to extreme temperatures. This is the reason why
civet coffee producers in Bengkulu prefer to use this type of coffee as a basic ingredient for civet
coffee. The optimal temperature for Robusta coffee growth is 21-24 0C.
Civet coffee production on a large scale causes problems for the civet population. Many civets are
hunted to be used as civet coffee-producing animals. this directly disrupts the mongoose population.
To cope with the growing demand for civet coffee, producers cannot only expect production from
mongoose animals. The utilization of mongoose animals as fermentation agents is considered to
torture mongoose animals as well as threatening the survival and preservation of mongoose animals.
One way to produce civet coffee without disturbing the mongoose animal population is to do
fermentation by using coarse isolate of mongoose coffee that comes from civet feces.
2. Methods
2.1. Research time and location
This research was conducted in November 2015 - March 2016 at the IHPT Laboratory, Argo
ecotechnology Laboratory, Chemistry Education Laboratory, Biomedicine Laboratory, Faculty of
Medicine, Bengkulu University and SBIH Ruyani Laboratory (Learning Nature Harmony From the
Facts).
2.2. Determination of Robusta coffee protein concentration profile
Microbes from civet feces mixed with Robusta coffee beans were incubated for 24 hours in a 0.9%
Physiological NaCl solution, forming a precipitate. The liquid which is at the top of the sediment is
mixed into the liquid Nurien Borth (NB) and incubated for 24 hours, the result of this incubation is
called the coarse isolate. The coarse isolates obtained are then made into dilution series variations
10-1–10-6. The smallest dilution series (10-6) is taken and implanted on PCA media. Planting is
carried out by the top planting method and incubated for 24 hours. The results obtained were then
calculated the number of microbial colonies using "Colony Counter" to determine the amount of
microbial concentration for each dilution series.
This fermentation process is done by soaking the Robusta coffee beans and a little flesh with
isolates from various concentration variants (2.91x108 CFU / mL; 2.91x107CFU / mL; 2.91x 106CFU
/ mL, 2.91x105 CFU / mL; 2, 91x104 CFU / mL). Then incubated according to a predetermined time
variation (24,36,48, and 60 hours). And stirring is done every 3 hours. After the fermentation process
is complete, Robusta coffee beans are washed with 70% alcohol and then rinsed with distilled water.
Protein isolation is done by grinding fermented coffee beans, then homogeneous using Tris-HCL
buffer pH 7.4 for 24 hours. Next homogenate is filtered with filter paper and centrifuged. The
centrifugation process is carried out in 2 stages, the first is carried out at 4.500 rpm for 15 minutes and
the second stage is carried out at 13.500 rpm for 30 minutes to separate the precipitated protein by
dissolution, the pellet is taken because it is a precipitation protein, the determination of protein
concentration from the pellet will be carried out.
The determination of protein concentration was carried out using a Uv-vis spectrophotometer at a
wavelength of 570 nm. The method of determining the concentration of protein used is the Biuret
method using a standard protein BSA (Bovin Serum Albumin). BSA solution is made in a biuret
solution with various concentrations of 20 μg / mL, 40 μg / mL, 60 μg / mL, 80 μg / mL, 100 μg / mL.
4. Mathematics and Science Education International Seminar (MASEIS) 2019
Journal of Physics: Conference Series 1731(2021) 012019
IOP Publishing
doi:10.1088/1742-6596/1731/1/012019
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2.3. Determination of Robusta coffee protein molecular weight profile
The electrophoretic gel is made for bottom-PAGE and upper-PAGE with a concentration of 12%. The
gel is allowed to stand for several minutes. Chip with some wells that are ready to be filled with
protein samples. The protein standards used in this analysis are broad range prestained SDS-PAGE
BIO-RAD with the following criteria: myosin (210KDa), β-Galactosidase (125KDa), bovine serum
albumin (101KDa), Ovalbumin (56.2 KDa) Carbonic anhydrase (35.8KDa), soybean trypsin inhibitors
(29 KDa), lysozyme (21 KDa), aprotinin (6.9 KDa).
Electrophoresis was carried out at a constant voltage of 220 V and stopped when the tracer colour
(bromophenol blue) had moved to reach the lower end of the gel or ± 60 minutes. The 1-D PAGE
produced was then coloured with Coomassie Bio-safe [6]. Protein bands are observed and
photographed directly using a digital camera. The number of components is determined by the number
of stains obtained from the separation results. Then the molecular weight is analysed by comparing the
protein band in the sample with the standard protein band.
2.4. Data analysis techniques
Data analysis techniques for laboratory experiments were carried out based on the results of
calculations of protein concentrations contained in the samples used. The concentration of this protein
can be determined by calculating the absorbance value shown by the Uv-Vis spectrophotometer. The
protein concentrations obtained were then analysed using Microsoft Excel 2010. In this research, a real
difference test was performed using the parametric statistical analysis of variance (ANOVA) test,
namely the Randomized Group Design (RCBD) and continued with the CRD (Randomized Complete
Design).
3. Result and discussion
3.1. Protein concentration profile of Robusta coffee
Based on the results of this research showed that the Robusta coffee protein concentration decreased to
166.67 μg / mL when compared with Robusta coffee fermented in vitro (fermentation time of 60 hours
and microbial concentrations of rough isolates 2.91 x 108 CFU / mL). this result is different when
compared with in vivo fermented civet coffee, the concentration of pure Robusta coffee protein has
decreased to 50.17 μg / mL. Average in vitro fermentation concentration data for each concentration
and time variation can be plotted in Figure 1:
Figure 1. The average concentration curve of In Vitro fermentation.
5. Mathematics and Science Education International Seminar (MASEIS) 2019
Journal of Physics: Conference Series 1731(2021) 012019
IOP Publishing
doi:10.1088/1742-6596/1731/1/012019
4
Based on Figure 1. it can be seen that the lowest protein concentration of Robusta coffee beans in vitro
is 50.95 μg / mL (fermentation time is 60 hours and the concentration of crude isolates is 2.91 x108
CFU / mL). Based on the curve it can be seen that the longer the fermentation time and the greater the
concentration of civet coffee isolates used, the concentration of Robusta coffee protein will be lower.
This is because the longer the fermentation time, the more content of substances used by bacteria to
survive so that the amount of food remaining less and less including protein compounds.
As for the concentration of microbes, the greater the concentration of microbes in coarse isolates,
the greater the concentration of protein which decreases, this is due to the large amount of microbial
concentration that works to reduce the protein in coarse isolates so that the content of substances used
by microbes to survive will be more numerous, so that food substances the remaining will be less so as
well as protein compounds [7]. The longer the fermentation time, the longer bacteria will decompose
the compounds contained therein such as sugar, protein, cellulose [8].
Several factors that influence microbial growth are the availability of nutrients, water, temperature,
pH, oxygen, reduction oxidation potential, the presence of inhibitors, and the presence of other
microorganisms [9]. Differences in growth in microbes are caused by physiological diversity and
different responses to physical conditions and the environment [10].
Robusta coffee with low protein concentration is indeed the coffee desired in this research because
coffee with low protein concentration has a delicious taste and has a bitter taste. Protein generally acts
as a form of bitter taste in roasted coffee so that civet coffee is not as bitter as ordinary coffee because
of its low protein content. This is consistent with the opinion of Marcone [4] improvement in the
quality of civet coffee flavour caused by a low protein content and high fat content compared to
ordinary coffee. Low protein content can reduce bitter taste, while high fat content can increase body.
In addition to reducing the bitter taste in coffee, low protein content in civet coffee can also support a
diet program without protein.
3.2. Molecular weight profile of Robusta coffee
Determination of the molecular weight of Robusta coffee protein in vitro fermentation was carried out
using electrophoresis. Electrophoresis is a protein separation technique based on the movement of
charged protein molecules in an electric field (isoelectric point). The speed of molecules moving in the
electric field depends on the charge, shape and size. Electrophoresis results can be seen in the image
below:
Figure 2. Gel electrophoresis 1. Figure 3. Gel electrophoresis 2.
Based on figures 2 and 3, the higher the concentration of coarse isolates used in the fermentation
process, the colour of the protein bands produced in the electrophoretic gel will be thicker and the
number of bands produced will be more than that of samples fermented with isolates that have lower
concentrations. This occurred in almost all samples of Robusta coffee protein fermented in vitro,
6. Mathematics and Science Education International Seminar (MASEIS) 2019
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doi:10.1088/1742-6596/1731/1/012019
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except for Robusta coffee protein samples with an isolate concentration of 2.91 x 108 CFU / mL and
isolate concentration of 2.91 x 108 CFU / mL with 48 hours fermentation time (Figure 1). This is
thought to occur because the Robusta coffee protein has been completely hydrolysed into amino acids
during the fermentation process.
Samples with high isolate concentrations generally form more bands during the electrophoresis
process. This shows that the hydrolysis process is going quite well, although not perfectly. This means
that most protein molecules are hydrolysed into amino acids, but some are not. Proteins that are
hydrolysed into amino acids will later form many bands on the electrophoretic gel according to their
respective molecular weights.
Figures 2 and 3, several Robusta coffee protein samples only have one band on the top of the
electrophoresis gel. The protein band at the top of the electrophoresis gel is a large molecular weight
protein, a protein with a large molecular weight will not be able to go down to the bottom of the well,
because the protein down to the bottom of the well is a protein with smaller molecular weight.
The fermentation time did not significantly influence the results of the research. This can be seen
where the protein band produced does not decrease based on the length of time of fermentation, but
decreases along with the decline in the concentration of microbes used in the fermentation process for
each time of fermentation. This might be due to the imperfect fermentation process or it could also be
caused by protein denaturation before the fermentation process is carried out.
3.3. The linkages between the results of the analysis of concentration profiles and the molecular
weight profile of Robusta coffee proteins
The results obtained from this research indicate that the results of the analysis of protein concentration
profiles (Figure 1) are not always directly proportional to the results of the protein molecular analysis
weight profiles in Robusta coffee (Figures 2 and 3). This can be seen from protein samples with
(Luwak coffee isolate concentration 2.91 x 108 CFU / mL and isolate concentration 2.91 x 107 CFU /
mL with fermentation time 48 hours) which has a high protein concentration value based on the results
of the UV-vis spectrophotometer, but it does not appear on the molecular weight band on the
electrophoretic gel. It is suspected, the protein read by the UV-Vis spectrophotometer is the total
protein of all types of proteins, including protein fractions (oligoproteins).
The absence of protein bands in electrophoresis process, may be caused by the undetectable
amount of protein as a whole, which is thought to be only protein with a molecular weight that is not
too small that can form a band on the electrophoresis gel, whereas for proteins with very small
molecular weights as in the form of oligoproteins or even in the form of fractions, cannot form a band
because it is suspected that when the electrophoresis process takes place this type of protein molecule
has come down perfectly to the part that the well, making it difficult to detect even though the process
of purification and colouring in the gel has been carried out.
The purity of the analysed protein also seems to be one of the gaps in the results obtained. The
coffee protein analysed is thought to be not purely Robusta coffee protein but has been mixed with
proteins from proteolytic bacteria and proteins from protease enzymes. When the process of protein
hydrolysis takes place, it is suspected that the protease enzyme derived from proteolytic bacteria and
proteolytic bacteria itself also hydrolyses along with proteins from coffee beans. As a result, the
protein analysed both on the spectrophotometer and with electrophoresis is a mixture of protein from
coffee bean protein, protease enzyme protein, and protein from proteolytic bacteria. It is strongly
suspected that the band at the top of the electrophoresis gel is a Robusta coffee protein sample band,
while the protein band that appears irregularly in the middle and bottom of the electrophoresis gel is
thought to be a protein derived from proteolytic bacteria and protease enzymes that have undergone
hydrolysis in during the fermentation process.
The thickness of the polish of electrophoresis gel (the blue colour that extends along the
electrophoresis well) produced by each protein sample is different, depending on the concentration of
protein remaining after the running process is complete. Samples that have a thicker polish colour are
thought to have more protein concentrations when compared to samples that have a thinner polish
7. Mathematics and Science Education International Seminar (MASEIS) 2019
Journal of Physics: Conference Series 1731(2021) 012019
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doi:10.1088/1742-6596/1731/1/012019
6
colour. This has a connection with the fermentation process, in which a properly hydrolysed protein
will go down the gel completely when the running process is carried out, so that the protein
concentration in the gel well is reduced or even absent so that when reacted with Coomassie blue, the
resulting blue colour is not so thick or even does not appear, whereas in samples with BM it is likely
that when the running process is complete there is still a lot of protein concentration in the gel well so
that when Coomassie blue is added, the protein will react with Coomassie blue to form a blue colour
along the gel electrophoresis well.
4. Conclusion
Fermentation time and concentration of coarse civet coffee isolates are inversely proportional to the
Robusta coffee protein concentration. Whereas the concentration of Luwak Robusta coffee isolates
used in the fermentation process is directly proportional to the appearance of protein sample bands on
the resulting electrophoretic gel. While the fermentation time did not have a significant effect because
no significant differences were found between each sample with different time variants.
Acknowledgments
The author would like to thank all those who have helped in completing this paper, especially to
Universitas Bengkulu and the SBIH Ruyani laboratory.
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