• Asian-Australasian Journal of Animal Sciences
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• v.16 no.5
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• pp.677-684
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• 2003

Twenty eight multiparous lactating cows were utilized in an experiment to evaluate the response to an exogenous fibrolytic enzyme on their lactational performance during early lactation period (in terms of milk production, milk composition, feed intake, milking efficiency, body weight change) and the exact time of this response. Cows were randomized into two groups (14 each) with similar parities and were fed a concentrate ration of barley, ground corn, soybean meal, and wheat bran and roughage ration of alfalfa hay. One of the two groups was supplemented with the fibrolytic enzyme immediately after parturition up to 100 post partum. The experiment was of two phases with 50 days each. The enzyme, which has a cellulase/hemicellulase activity (derived from Trichoderma group), was added to the concentrate part of the ration in a dry powder form. Milk production, 3.5% fat corrected milk, energy corrected milk were higher (p<0.05) for cows fed treated diet. At the same time, No differences were observed in percentages of milk components, feed intake, body weight, body weight change, or rectal temperature for the whole experimental period or during any of the two phases. Efficiency of milk production was higher (p<0.05) for treatment group cows than for that of the control ones. However, efficiency was better during the second phase than during the first phase. Feeding enzyme treated diets to dairy cows improved lactational performance during early 100 day of the lactation period. However, the first 50 days of lactation looked to be the critical.

• Korean Journal of Food Science and Technology
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• v.3 no.3
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• pp.172-177
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• 1971

The amount of the milk clotting enzyme which is produced by Dothiorella ribis in wheat bran was 950 Soxhlet units per gram of wheat bran. The milk clotting activity of this enzyme was increased by the elevation of clotting temperature and by the increase of the addition of $CaCl_2$ to milk. It was also increased when the pH of milk was lower than that of the fresh milk. When milk was diluted by distilled water, the milk clotting activity of the enzyme was decreased. And its milk clotting activity was good when milk was pasteurized at low temperature. The enzyme of Dothiorella ribis has larger proteolytic activity per Soxhlet unit than that of the milk clotting enzyme of Mucor pusillus Lindt. This enzyme was rather stable between pH 6 and pH 8 when it was conditioned for ten minutes. The heat stability of enzyme was tested by treating it under the condition for $10{\sim}60$ minutes. And the enzyme was stable under the temperature of $45^{\circ}C$. Also the recovery of protein as a form of curd was 76.2 percent to the total protein content of milk.

• Journal of Animal Science and Technology
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• v.45 no.1
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• pp.131-142
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• 2003

We evaluated the effects of adding fibrolytic enzyme into ruminant diets on ruminal fermentation (in vitro) and lactational performances of dairy cows (in vivo). Through the in vitro experiment that was carried out with different contents of NDF (34, 38, 43%) in diets, digestibilities of NDF in the rumen appeared not significantly different by the addition of enzyme but were different by NDF content in diets showing higher digestibility in NDF 43% diet. It could be attributed by the relatively higher amount of hemicellulose in the current experimental diets than in conventional diets that might have been digested easily by the addition of fibrolytic enzyme in the rumen. The addition of fibrolytic enzyme tended to increase NDF digestibilities to a little extent both in 0.05 and 0.1% enzyme levels. Ruminal pH, NH3-N concentrations and VFA production in the rumen were not affected by the addition of fibrolytic enzyme. Activities of CMCase and xylanase were higher in enzyme treated diets of both NDF 34 and 38%. In particular, the activities of xylanase that slowly decreased from 0 to 12 hr but rapidly after 24 hr indicates that the major action of the enzyme in the rumen occurs in early period of incubation. Through an in vivo experiment, fibrolytic enzyme addition into the diets of dairy cows indeed affected lactational performance of milk yield. The cows fed enzyme treated diets produced 8% (1.9kg/d) more amounts of milk than with no enzyme addition. Milk composition of milk fat and protein was not affected by enzyme addition. Overall, the results of this in vivo study indicates that fibrolytic enzyme can be used to improve milk production in lactating cows. In respect that animals in different treatments of this study had the same amounts of intake, the increased milk yield with enzyme addition may be attributed to the improved utilization of nutrients in the digestive tract.

• Archives of Pharmacal Research
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• v.17 no.5
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• pp.318-322
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• 1994

Upon gel filtration, the commercial bovine milk xanthine oxidase preparation was fractionated into two preparations showing enzyme activity. Native polyacrylamide gel electrophoresis showed that one was in a dimeric form and the other was a monomer having molecular weight of 150 kDa. It was also found that this commercial enzyme existed mostly in an active monomeric form without loss of enzyme activity. The rabbit antisera produced against two enzyme preparations cross-reacted well each other. In SDS-polyacrylamide gtel electro-phoresis, however, both enzyme preparations yielded two smaller protein bands below 150 kDa, which appeared to bind with both antisera with high affinity but not to retain enzyme activity. It implies that bovine milk xanthine oxidase can lose its activity when monomeric subunit is further degraded.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.4
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• pp.418-422
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• 1992

Mucor mucedo C-7 selected as a potent fungus for producing milk-clotting enzyme was cultured on wheat bran solid medium and the optimum culture conditions for the production of milk clotting enzyme were ot tamed as follows. Amount of water added to wheat bran was 100% to the weight of wheat bran and culture temperature and time was 3$0^{\circ}C$ and 72hrs, respectively. The production of milk-clotting enzyme was markedly increased by the addition of Macllvaine buffer solution (pH4.5) instead of water added to wheat bran solid medium and milk-clotting activity was stable for culture period.

• Microbiology and Biotechnology Letters
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• v.19 no.2
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• pp.109-115
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• 1991

Twenty-five fungal strains producing an extracellular soybean milk clotting enzyme were isolated from 146 soil samples, and identified as 11 species belonging to six genera of Aspergillus oryzae (5 strains), Aspergillus flavus (2 strains), Aspergillus parasiticus (1 strain), Aspergillus tamarii (2 strains), Aspergillus niger (4 strains), Aspergillus fumigatus (2 strains), Mucor hiemalis (2 strains), Wallemia sebi (4 strains), Scopulariopsis condida (1 strain), Fusarium redolens(1 strain) and Verticillum lecanii (1 strain). Among them, Aspergillus oryzae 020 and Aspergillus tamarii 287 showed relatively high soybean milk clotting activity. The coagulabilities of the enzyme from representative strains of those species decreased as the pH of soybean milk increased from 6.0 to 7.0 The optimum temperature for soybean milk clotting enzymes of those strains were 65$^{\circ}C$.

• Applied Biological Chemistry
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• v.12
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• pp.7-11
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• 1969

Mucor-rennin, the crystalline milk-clotting enzyme, isolated from Mucor pusillus var. Lindt, has an acid protease activity. The optimum pH for the digestion of k-casein is 4.5, while that for hemoglobin digestion is 4.0. The skim milk solution was easily clotted acidic solution than alkalin solution, and the milk clotting activated by Ca ion. The enzyme was heat stable against heat from pH 4.0 to 6.0 but was more stable at pH 5.0. The activity of the enzyme at pH 5.0 did not decrease at 30 C for 15 days and the activity was not effected by sodium propionate and salicilic acid. Therefore, the enzyme of liguid type could store for a long time and could be transported from Erzyme production Co. to Manufacture of cheese Co. by adding the antiesptic and by adjusting pH to 5.0.

• Microbiology and Biotechnology Letters
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• v.17 no.2
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• pp.109-114
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• 1989

Seventeen bacterial strains producing an extracellular soybean milk clotting enzyme were Isolated from 150 soil samples, and identified as Bacillus cereus(8 strains), Bacillus pumilus(8 strains) and Bacillus licheniformis (1 strain). Among them, Bacillus pumilus strain 118 and Bacillus licheniformis strain 192 showed relatively high soybean milk clotting activity. The coagulability of enzymes from these strains decreased as the pH of soybean milk was increased from 6.0 to 7.0. The optimum temperature for soybean milk clotting activity was $65^{\circ}C$.

• Korean Journal of Agricultural Science
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• v.14 no.1
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• pp.144-155
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• 1987

A potent fungus producing milk clotting enzyme with fairly weak proteolytic activity was isolated from various soil and sewage, which the selected strain, SA-101, was identified as Mucor sp. with microbiological characteristics. Its milk clotting enzyme production was maximized when grown on 10g of wheat bran media added to 8ml of tap water containing 0.1M HCl for 60hrs at $30^{\circ}C$. This enzyme production was stimulated by addition of 6% lactose, 0.05% NaCl and reached a maximal level of 9810 unit/g wheat bran. The crude enzyme product could be produced effectively by salting out with ammonium sulfate fractionation and lyophilization. The ratio of milk clotting activity to proteolytic activity of crude enzyme product was lower than Hansen rennet, but resembled to Meito rennet. The optimal temperature of milk clotting activity of crude enzyme product was abound $60^{\circ}C$ on a substrate of 10% reconstituted skim milk containing 1/100M $CaCl_2$.

• Journal of Microbiology and Biotechnology
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• v.23 no.8
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• pp.1084-1091
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• 2013

In this paper, two statistical methods were applied to optimize medium components to improve the production of the milk-clotting enzyme by Bacillus amyloliquefaciens D4. First, wheat bran juice, skim milk powder, and $Na_2HPO_4$ were shown to have significant effects on D4 enzyme production using the Plackett-Burman experimental design. Subsequently, an optimal medium was obtained using the Box-Behnken method, which consisted of 3.31 g/l of skim milk powder, 5.0 g/l of sucrose, 0.1 g/l of $FeSO_4{\cdot}7H_2O$, 0.1 g/l of $MgSO_4{\cdot}7H_2O$, 0.1 g/l of $MnSO_4{\cdot}2H_2O$, 0.1 g/l of $ZnSO_4{\cdot}7H_2O$, 1.52 g/l of $Na_2HPO_4$, and 172.45 g/l of wheat bran juice. With this optimal medium, the milk-clotting enzyme production was remarkably enhanced. The milk-clotting enzyme activity reached 3,326.7 SU/ml after incubation of 48 h, which was 1.76-fold higher than that of the basic medium, showing that the Plackett-Burman design and Box-Behnken response surface method are effective to optimize medium components, and B. amyloliquefaciens D4 possessed a high rennet-producing capacity in the optimal medium.