Studies on the Production of Acid Protease by Submerged Fermentation

2009 ◽  
Vol 5 (1) ◽  
Author(s):  
Shishir Sinha ◽  
Shalini Sinha
Keyword(s):  
Enzyme Activity ◽  
Proteolytic Activity ◽  
Enzyme Production ◽  
Milk Powder ◽  
Skim Milk ◽  
Peanut Meal ◽  
Tween 80 ◽  
Skim Milk Powder ◽  
Acid Protease ◽  
Aspergillus Awamori

Proteases are enzymes that hydrolyze peptide bonds and, therefore, lead to the disassembly of proteins. Commercially these are extremely important as more than 60% of the total enzyme market is made up of proteases, out of which 40% are acid proteases. The objective of this study was to compare the available standard strains, Rhizopus oligosporus MTCC-556, Rhizomucor miehei MTCC-546 and Aspergillus awamori MTCC-548, which were examined for the production of acid protease by submerged fermentation. Aspergillus awamori showed maximum proteolytic activity and was selected for further optimization studies. During the course of study the medium was altered. Effect of different carbon sources (lactose, sucrose, and combination of these two in same ratios and glucose) on the proteolytic activity of acid protease produced by A. awamori MTCC 548 was studied and it was found that glucose showed highest proteolytic activity. The effect of various concentrations of glucose was also studied on the acid protease production and its 1% concentration was found to be optimum; it showed proteolytic activity of 0.11 U/ml. Among the different nitrogen sources, such as casein, peptone, skim-milk powder and peanut meal, the peanut meal was found better for enzyme production. Peanut meal, with a concentration of .2% in the medium, increased proteolytic activity up to 0.218 U/ml. The effect of additives, such as Tween-80 and chemicals like CaCl2 and skim-milk powder, was also studied and it was found that 0.05% Tween-80 was effective in enzyme production and a proteolytic activity of 0.225 U/ml was obtained. The enzyme extract was separated in the form of supernatant by using centrifuge and the enzyme activity was analyzed by Anson’s method using a spectrophotometer. The enzyme produced was recovered by using a saturated solution of 80% ammonium sulphate and protein content was determined by Lowery method using a spectrophotometer. It was found that the specific enzyme activity was increased from 0.155 U/mg proteins to 0.174 U/mg proteins showing a purification fold by a factor of 1.12 by using 80% saturated ammonium sulphate.

Concurrent studies of the flow of digesta in the duodenum and of exocrine pancreatic secretion of calves

1974 ◽  
Vol 31 (1) ◽  
pp. 13-26 ◽  
Author(s):  
J. H. Ternouth ◽  
J. H. B. Roy ◽  
R. C. Siddons
Keyword(s):  
Enzyme Activity ◽  
Pancreatic Secretion ◽  
Pancreatic Enzyme ◽  
Milk Powder ◽  
Skim Milk ◽  
Skim Milk Powder ◽  
Protein Nitrogen ◽  
Postprandial Period ◽  
Pancreatic Protease ◽  
Protease Secretion

1. The duodenal flow of digesta and the concurrent pancreatic secretion were compared when six Ayrshire calves, with duodenal re-entrant and pancreatic sac cannulas, were fed on three reconstituted milks. The diets were: reconstituted, ‘mildly’ preheated, spray-dried skim-milk powder (SK); the same skim milk containing 20 g fat/l (SKF); and reconstituted, ‘severely’ preheated skim-milk powder containing 20 g fat/l (HSKF). The calves were fed ad lib. from teats twice daily from 9 to 21 d of age, each diet being offered for 4 d. Collections of duodenal digesta and pancreatic secretions were made for 12 h after the fourth and eighth meals on each diet.2. The calves tended to have the highest liquid intakes when diet SK was given. After adjustment for differences in intake, diet SK resulted in the appearance of more hydrogen, chloride and potassium ions but less undigested protein nitrogen in the duodenal digesta than with either diet SKF or HSKF.3. Compared with diets SK and SKF, the whey fluids from diet HSKF took significantly longer to leave the abomasum, less H+ passed through the duodenum during the first 6 h after feeding and less Cl− during the whole postprandial period. More undigested protein N and fat from diet HSKF passed through the duodenum during the first 6 h after feeding, although this difference was significant only for protein N during the 1st hour after feeding.4. Over the 12 h postprandial period, the duodenal digesta contained almost exactly the same quantities of polyethylene glycol (PEG), N and fat as those in the meal. The total volume of digesta was 2.25 l greater than the quantity of milk ingested. When the hourly duodenal flows of PEG and fluid were expressed as the square root of the hourly quantities recovered, the pattern of abomasal emptying was rectilinear. The flows of N and fat were curvilinear, when expressed on the same basis.5. The concentration of ‘sodium-free’chloride in the duodenal digesta, in excess of that ingested in the milk, was used as an indicator of the quantity of acid secreted by the abomasum. The relative quantity of acid secreted was greatest with diet SK and least with diet HSKF.6. The pancreatic secretion of fluid was highest during the period 5–9 h after feeding but the secretion of enzyme activity was highest during the first 2 h after feeding.7. Considerable variability in the secretion of enzyme activity was observed and the rate of secretion did not appear to be related to any component of the duodenal digesta.8. Diet SKF was associated with a greater volume of pancreatic secretion and more pancreatic protease secretion than either diet SK or HSKF, but most amylase activity was secreted when diet HSKF was given. Evidence is presented which suggests that pancreatic enzyme activity adaptation occurred when diet HSKF was offered in succession to diet SK or SKF. The secretion of trypsin activity did not differ between diets.

scholarly journals Concurrent studies of the flow of digesta in the duodenum and of exocrine pancreatic secretion of calves

1975 ◽  
Vol 33 (2) ◽  
pp. 181-196 ◽  
Author(s):  
J. H. Ternouth ◽  
J. H. B. Roy ◽  
S. Y. Thompson ◽  
Joyce Toothill ◽  
Catherine M. Gillies ◽  
...  
Keyword(s):  
Enzyme Activity ◽  
Pancreatic Enzyme ◽  
Milk Powder ◽  
Skim Milk ◽  
Milk Proteins ◽  
Skim Milk Powder ◽  
Protein Marker ◽  
Total N ◽  
Postprandial Period ◽  
Pancreatic Fluid

1. The flow of digesta through the duodenum and the concurrent secretion of the pancreas were studied in four Friesian calves given four milk-substitute diets. The diets were: reconstituted, ‘mildly’ pre-heated, spray-dried skim-milk powder with (SKF) or without (SK) margarine fat or with 50% of the skim-milk powder in diet SKF replaced by soya-bean flour (ASKF) or fish-protein concentrate (BSKF), together with dried whey. The diets were given ad lib. twice daily from 13 to 37 d of age, each diet being given for 6 consecutive days. Collections of duodenal digesta and pancreatic secretions, from cannulas, were made for 12 h after feeding the 6th and 12th meals (‘experimental’ meals) for each diet2. The diets fed as ‘experimental’ meals contained polyethylene glycol (PEG) as a fluid (whey) marker and goat's milk containing [3H]lysine as a marker for total protein; β-carotene was added as a lipid marker to the three diets containing margarine fat3. Over the 12 h postprandial period, the patterns of duodenal digesta flow and secretion of pancreatic fluid did not differ markedly between the four diets. The abomasal outflow of both nitrogen and lipid in a 12 h postprandial period was related to their intakes from the ‘penultimate’(5th and 11th) meals for diets SKF and SK but to their intakes at the ‘experimental meals’ for diets ASKF and BSKF. Secretion of pancreatic enzyme activity was highest during the 1st hour after feeding but the main outflow from the abomasum of total N and lipid occurred 5–10 h after feeding4. The time required for all the whey marker (PEG) to pass through the duodenum was similar for diets SKF and SK, but only 53 and 42% respectively of the ingested protein marker passed through the duodenum in the 12 h after feeding. More acid appeared to be secreted by the abomasum when diet SK was given; also less undigested protein passed out of the abomasum after giving this diet. It is concluded that the physical absence of fat globules in the abomasal clot increases the degree of proteolysis5. The secretions of pancreatic fluid and pancreatic enzyme activity were all markedly lower for diet SK than for diet SKF6. With diets containing non-milk proteins (ASKF and BSKF), abomasal proteolysis was less efficient and the ingested protein passed out of the abomasum more rapidly than for diet SKF. There was no difference in the rate of abomasal outflow of the whey fluids between diets SKF, ASKF and BSKF7. In comparison with diet SKF, diets ASKF and BSKF tended to induce less pancreatic enzyme secretion over a 12 h postprandial period, with the exception of lipase8. There appeared to be no direct relationship between the quantities of any of the pancreatic enzymes secreted during a postprandial period and either the concurrent flow of duodenal digesta or the total quantities of dietary constituents passing through the duodenum.

scholarly journals Irinotecan Engineered Proniosomes: In vitro and In vivo Characterization

2021 ◽  
pp. 230-238
Author(s):  
Prakash Goudanavar ◽  
B. Ramesh ◽  
Santosh Fattepur ◽  
Nagaraja Sreeharsha
Keyword(s):  
In Vitro Release ◽  
Milk Powder ◽  
Skim Milk ◽  
Tween 80 ◽  
Pharmacokinetic Profile ◽  
Skim Milk Powder ◽  
Tween 20 ◽  
Acceptable Method

Objective: The focus of this research has been to improve efficacy, decrease tolerance and increase the irinotecan pharmacokinetic profile. Methods: Proniosomesformulated with various surfactants, cholesterol and dicetyl phosphate using the slurry method. A slurry process was used to prepare proniosomes with maltodextrin as the carrier by using surfactants span 20, span 60, tween 20 and tween 80. Results: The preparations were characterized in terms of shape and specific surface area, entrapment efficacy, in vitro release studies, in vivo tissue diffusion and stability testing. The proniosome surface was found to be smoother in nature showing thin and compact layer with skim milk powder. For formulation 2 (73.94±2.8%), the maximum entrapment efficacy was found. Conclusion: The formulation 3 obtained the desired maximum release profile within 24 hours (98.06%). The in vivo tissue distribution studies for the proniosomes reveal that the drug was preferentially targeted to liver followed by the alveolus and lymphatic system.Stability studies have indicated that the most acceptable condition for storage of the formulation 2 was 4o C. Proniosomes provide an acceptable method to the carrier for targeted therapy. These can be held at specific sites and can release the drug for a prolonged period of time.

Improved heat stability of recombined evaporated milk emulsions by wet heat pretreatment of skim milk powder dispersions

2021 ◽  
pp. 106757
Author(s):  
Jianfeng Wu ◽  
Simin Chen ◽  
Teng Wang ◽  
Hao Li ◽  
Ali Sedaghat Doost ◽  
...  
Keyword(s):  
Milk Powder ◽  
Skim Milk ◽  
Heat Stability ◽  
Skim Milk Powder ◽  
Heat Pretreatment ◽  
Wet Heat

Anti-obesity effects of Lactobacillus rhamnosus 4B15, and its synergy with hydrolysed lactose skim milk powder

2021 ◽  
pp. 104997
Author(s):  
Sejeong Kim ◽  
Jae Yeon Joung ◽  
Daekyoung Kang ◽  
Nam Su Oh ◽  
Yohan Yoon
Keyword(s):  
Milk Powder ◽  
Lactobacillus Rhamnosus ◽  
Skim Milk ◽  
Skim Milk Powder

A note on the performance and management of calves reared on cold acidified milk replacer fed ad libitum

1983 ◽  
Vol 36 (1) ◽  
pp. 147-150 ◽  
Author(s):  
W. S. Thickett ◽  
N. H. Cuthbert ◽  
T. D. A. Brigstocke ◽  
M. A. Lindeman ◽  
P. N. Wilson
Keyword(s):  
Milk Powder ◽  
Skim Milk ◽  
Live Weight ◽  
Skim Milk Powder ◽  
Barley Straw ◽  
Milk Replacer ◽  
Bull Calves ◽  
The Mean ◽  
Ad Libitum ◽  
Dry Food

ABSTRACTResults are presented from six trials dealing with aspects of management on the cold ad libitum system of calf rearing using an acidified milk replacer containing over 600 g skim milk powder per kg.Thirty-six calves were housed in pens of six for each trial and were fed through a teat and pipeline from a storage barrel. Acidified milk replacer, pH 5·6, was mixed cold at 125 g/1 and made available ad libitum to 3 weeks. A rationed allowance was given daily, on a reducing scale, over the following 2 weeks with weaning completed at 35 days. A pelleted dry food containing 180 g crude protein per kg, together with water in buckets and barley straw in racks, was available ad libitum throughout. Each trial lasted 8 weeks. Results for the mean of the six cold ad libitum trials involving 216 calves were compared with the mean results of 10 conventional bucket-fed trials carried out separately at the same unit, involving 912 calves. All calves were purchased British Friesian male (bull) calves.Calves on the ad libitum system showed improved live-weight gains of 9·4 kg at 3 weeks, 8·8 kg at 5 weeks and 7·5 kg at 8 weeks, compared with the conventional system. The consumption of milk replacer powder was higher in ad libitum trials at 29·4 kg cf. 12·5 kg by bucket but intake of pelleted dry feed was lower on the ad libitum system at 50·7 kg cf. 71·3 kg to 8 weeks. Calf appearance scores were significantly improved on the ad libitum system which gave the main improvement in performance in the first 3 weeks.

scholarly journals Staphylococcal Food Poisoning Associated with Spray-Dried Milk

1955 ◽  
Vol 53 (4) ◽  
pp. 387-397 ◽  
Author(s):  
P. H. R. Anderson ◽  
Doris M. Stone
Keyword(s):  
Spray Drying ◽  
Clinical Features ◽  
Milk Powder ◽  
Food Poisoning ◽  
Skim Milk ◽  
Skim Milk Powder ◽  
Large Numbers ◽  
Heat Treated ◽  
Drying Chamber ◽  
Spray Dried

SummaryEight explosive outbreaks of food poisoning, occurring in school canteens in England during 1953 and affecting 1190 known cases, are described. The clinical features were characteristic of the toxin type of illness. No deaths occurred.The food causing all of these outbreaks was prepared from spray-dried skim milk powder. It was not subsequently heat-treated and was usually consumed 3–4 hr. after preparation.The spray-dried milk powder proved to contain a high content of bacteria, including large numbers of Staph. aureus, of a phage pattern often associated with food poisoning. The assumption was therefore made that these outbreaks were caused by staphylococcal enterotoxin.Because the food was often consumed within 3–4 hr. of reconstitution of the milk powder—before, in fact, the staphylococci had had time to grow—it is concluded that the poisoning must have been due mainly to pre-formed toxin.Consideration is given to the opportunities for the formation of toxin in a spray-drying plant, and reasons are brought forward for believing that it is formed mainly in the balance tank where the warm milk is kept, sometimes for several hours, before passing into the final drying chamber.The processing of the milk and the precautions for preventing contamination of the finished product are discussed.

Probiotic Fermented Food Mixtures: Possible Applications in Clinical Anti-Diarrhoea Usage

1998 ◽  
Vol 12 (2) ◽  
pp. 97-105 ◽  
Author(s):  
Binita Rani ◽  
N. Khetarpaul
Keyword(s):  
Milk Powder ◽  
Skim Milk ◽  
Titratable Acidity ◽  
Experimental Period ◽  
Skim Milk Powder ◽  
Fermented Food ◽  
Shigella Dysenteriae ◽  
E Coli ◽  
Tomato Pulp ◽  
Food Mixture

A probiotic fermented PCMT food mixture was developed by fermentation of an autoclaved and cooled slurry of pearl millet flour, chickpea flour, skim milk powder and fresh tomato pulp (PCMT 2:1:1:1, w/w) with Lactobacillus acidophilus (105 cells/ml), a probiotic organism at 37°C for 24 h. Such a fermented mixture inhibited the growth of pathogenic organisms, namely Shigella dysenteriae, Salmonella typhosa and E. coli. A significant decline in pH with a corresponding increase in titratable acidity due to probiotic fermentation occurred in the developed food mixture. Feeding of the freshly developed fermented. mixture to mice suffering from E. coli induced diarrhoea, could help to arrest diarrhoea, reduce moisture, protein and ash contents in their faeces. The counts of lactobacilli increased whereas those of E. coli decreased remarkably in the faeces of mice from the 3rd day of the feeding trial till the end of experimental period. The beneficial effect of probiotic feeding may be due to antimicrobial substances produced by L. acidophilus, which might have neutralized the enterotoxins from E. coli. The cost of one 200 ml glass full of this probiotic drink is no more than one rupee.

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