• Journal of Animal Science and Technology
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• v.66 no.2
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• pp.326-339
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• 2024

This study was conducted to investigate the effects of protease supplementation and different nutrient density of diets in growing-finishing pigs. A total of one hundred-eight crossbred growing pigs ([Landrace × Yorkshire] × Duroc) with an initial body weight (BW; 18.74 ± 3.46 kg) were used for 15 weeks. Pigs were randomly assigned to six dietary treatments with 6 replicates of 3 pigs per pen in a 3 × 2 factorial through the following arrangement: Three groups of protease (1, Basal diets; 2, Protease A: 125 mg/kg protease derived from Streptomyces sps; 3, Protease B: 100 mg/kg protease derived from Bacillus licheniformis) at two different nutrient density diets (1, Basal requirement; 2, 0.94%-0.98% higher than requirement in dietary protein and 50 kcal/kg in energy). High nutrient (HN) diets showed higher average daily gain (ADG) (p < 0.05) and apparent total tract digestibility (ATTD) of crude protein (CP) (p < .0001) compared to basal nutrient (BN) diets during growing periods. Supplementation of protease showed higher BW (p < 0.05) and ADG (p < 0.05) compared to non-supplementation of protease during growing periods. Also, supplementation of protease showed higher ATTD of CP (p < 0.01), ATTD of gross energy (p < 0.05) and decreased blood urea nitrogen (BUN) level (p = 0.001) compared to non-supplementation of protease during finishing periods. Pigs which fed the protease showed decreased ammonia (NH₃) emissions (p < 0.05) during experiment periods and decreased hydrogen sulfide (H₂S) emissions (p < 0.01) during finishing periods. Interactions between nutrient density and protease were observed, which decreased the feed conversion ratio (p < 0.05) in HN diets without protease compared to BN diets without protease during weeks 4 to 6. Also, interaction between nutrient density and protease was observed, which resulted in improved ATTD of CP (p < 0.01) in response to PTA supplementation with HN diets during the finishing period. In conclusion, supplementation of protease reduces NH₃ in feces and BUN in whole blood by increasing the digestibility of CP and improves growth performance. Also, diets with high nutrient density improved growth performance and nutrient digestibility in growing periods.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.9
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• pp.1303-1313
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• 2017

Objective: This study evaluated the change and function of the pancreas, and small intestine in relation to growth performance of broilers on diets supplemented with raw soybean meal (RSBM) and protease. Samples of test ingredients and diets, after mixing and prior to being used were also assessed on contents of anti-nutritional factors. Methods: A $3{\times}3$ factorial study was used, with three levels of RSBM (commercial soybean meal [SBM] was replaced by RSBM at 0, 10%, or 20%) and protease (0.1, 0.2, or 0.3 g/kg). Each treatment was replicated six times with nine birds per replicate. Birds were housed in cages, in climate-controlled room and fed starter, grower and finisher diets. Results: Levels of trypsin inhibitors in the diets, containing varying levels of RSBM ranged between 1,730.5 and 9,913.2 trypsin inhibitor units/g DM. Neither RSBM nor protease supplementation in diets significantly affected (p>0.05) the body weight of broilers in the entire periods (0 to 35-d). Increasing the level of RSBM in diets increased the weight of the pancreas at d 10 (p<0.000), d 24 (p<0.001), and d 35 (p<0.05). Increasing levels of RSBM in the diets reduced the apparent ileal digestibility of crude protein (CP), and amino acid (AA) at d 24. Increasing level of RSBM in the diets decreased (p<0.01) pancreatic protein content, but this was increased (p<0.05) when protease was added to the diets (0 to 10-d). Increasing the level of protease improved the pancreatic digestive enzymes, including trypsin (p<0.05), chymotrypsin (p<0.01), and general proteolytic enzymes (p<0.05). Conclusion: The commercial SBM could be replaced at up to 20% by RSBM for broilers. Although protease supplementation slightly improved the digestive enzymes, and the ileal digestibilities of CP and AA, the CP and AA were negatively affected by increasing RSBM.

• Applied Biological Chemistry
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• v.15 no.1
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• pp.27-33
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• 1972

The alkaline protease was isolated from the culture material of monascus sp. on wheat bran culture. The crude purification of this enzyme was extracted with distilled water and precipitated with ammonium sulfate of 0.5 saturation. And, the activity of this enzyme was determind very strongly by folin's colorimetric method. The optimal pH of this enzyme was ranging from pH 10 to 13 and the optimal temperature was $50^{\circ}C$. The pH stability was ranging from pH 5 to 12 and the enzyme activity was not inactivated by heat treatment in lower temperature than $40^{\circ}C$. The enzyme was protected from heat denature by the treatment of $Pb^#$, $Ba^#$, $Co^#$, $Zn^#$, and $Cu^#$, but was inactivated with $Hg^#$, $Fe^#$ strongly. Moreover, one of these metal ions, the cupper ion, has a strong protective activity on enzyme heat denature. And, it was not effected by treatment of EDTA.

• Journal of Microbiology and Biotechnology
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• v.22 no.9
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• pp.1230-1236
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• 2012

Fish meal grades SL1 and SL2 from Sardine (Sardinella longiceps) and NJ from Pink Perch (Nemipterus japonicas) were evaluated as a sole source of carbon and nitrogen in the medium for alkaline protease production by Bacillus pumilus MTCC 7514. The analysis of the fish meal suggests that the carbon and nitrogen contents in fish meal are sufficient to justify its choice as replacement for other nutrients. Protease production increased significantly (4,914 U/ml) in medium containing only fish meal, compared with the basal medium (2,646 U/ml). However, the elimination of inorganic salts from media reduced the protease productivity. In addition, all the three grades of fish meal yielded almost the same amounts of protease when employed as the sole source of carbon and nitrogen. Nevertheless, the best results were observed in fish meal SL1 medium. Furthermore, protease production was enhanced to 6,966 U/ml and 7,047 U/ml on scaling up from flask (4,914 U/ml) to 3.7 and 20 L fermenters, respectively, using fish meal (10 g/l). Similarly, the corresponding improvement in productivities over flask (102.38 U/ml/h) was 193.5 and 195.75 U/ml/h in 3.7 and 20 L fermenters, respectively. The crude protease was found to have dehairing ability in leather processing, which is bound to have great environmental benefits.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.8
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• pp.1291-1300
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• 2018

Objective: This experiment was conducted to investigate the effects of dietary crude protein (CP) level and exogenous protease supplementation on growth performance, serum metabolites, carcass traits, small intestinal morphology and endogenous protease activity in broiler chickens reared under a tropical climate. Methods: A total of 480 day-old male broiler chicks were randomly assigned to eight dietary treatments in a $4{\times}2$ factorial arrangement. The main effects were CP level (21.0%, 19.7%, 18.5%, or 17.2% from 1 to 21 days and 19.0%, 17.9%, 16.7%, or 15.6% from 22 to 35 days) and protease enzyme supplementation (0 ppm or 500 ppm). All experimental diets were fortified with synthetic feed-grade lysine, methionine, threonine and tryptophan to provide the minimum amino acid recommended levels for Cobb 500. Results: Reducing dietary CP linearly reduced (p<0.05) growth performance, serum albumin, total protein, and carcass traits and increased (p<0.05) serum triglycerides and abdominal fat. There was no consistent effect of reducing dietary CP on morphological parameters of the intestine and on the pancreatic and intestinal endogenous protease activity (p>0.05). Protease supplementation improved (p<0.05) feed conversion ratio, body weight gain, carcass yield and intestinal absorptive surface area. Conclusion: Protease supplementation, as measured by growth performance, intestinal morphology and carcass yield, may alleviate the detrimental effects of low protein diets in broiler chickens.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.498-503
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• 1999

Protease produced from Bacillus sp. FSE-68 was isolated from Meju, a Korean fermented soybean starter, and was purified by a two-consecutive aqueous two-phase system. The change of partition coefficient (K) in the polyethylene glycol (PEG)/potassium phosphate buffer (PPB) aqueous two-phase system was measured at different pHs (6.0- 9.2), PPB concentrations (8-12%), and temperatures (4 and $20^{\circ}C$). As the PPB concentration in the aqueous two-phase system increased, the protease concentration in the top phase (PEG-rich phase) increased, thereby enhancing the partition coefficient. The minimum partition coefficient of the protease was achieved at pH 7.0, whereas that of the total protein was at pH 6.0. The biggest difference in partition coefficients of total protein and protease occurred at pH 6.0. It was interesting to note that the partition coefficient of protease decreased as the temperature increased. The optimum condition of the primary aqueous two-phase extraction of Bacillus sp. FSE-68 was pH 6.0, 14% (w/w) PPB, and 16% (w/w) PEG at $4^{\circ}C$, and the crude enzyme concentration in this system was 50% (w/w). The protease, which was concentrated in the top phase, was further mixed with 15% (w/w) PPB (pH 7.0) in the ratio of 1:1 at $20^{\circ}C$ to elute the bottom phase (PPB-rich phase). Using these steps, the purification fold achieved was 9.2 with a 44.7% yield.

• Microbiology and Biotechnology Letters
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• v.25 no.1
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• pp.51-55
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• 1997

To utilize the alkaline protease produced by Halomonas sp. ES-10 as an enzyme detergent, the crude enzyme was obtained by methanol precipitation and lyophilization. And it was processed to coated enzyme.The best mixing ratio of components such as coated enzyme, builders, actives, fillers and adjuvants on detergency was examined, and temperature and pH influencing detergency were also tested. Detergency test 0.15% detergent solution was carried out on EMPA test cloth #116 with shaking(90 rpm) for 10 min after 30 min of pretreatment. The detergent which contained coated-enzyme 1%, Zeolite 4A 20%, Tween 80 1. 5%, sodium borate 30%, sodium meta silicate 7.5% and water 40% showed about 90% of washing efficiency at 40$\circ $C and pH 10.0.

• Food Science of Animal Resources
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• v.35 no.2
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• pp.211-215
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• 2015

From the previous study, Enterococcus faecium SH01 was isolated from mukeunji, an over-ripened kimchi, and it produced bacteriocin SH01. Bacteriocin SH01 showed an inhibitory effect against Listeria monocytogenes ATCC 19111, a bacterial strain causing human listeriosis. Crude bacteriocin SH01 was purified by ammonium sulfate precipitation and its inhibitory activity at two concentrations (500 and 1,000 AU/g) against Listeria monocytogenes ATCC 19111 was investigated in ground beef at increasing temperatures (5, 10, 15, and 20℃) for 8 d. The number of Listeria monocytogenes ATCC 19111 significantly decreased (p<0.05) as the concentration of bacteriocin increased from 500 to 1,000 AU/g. Intrinsic crude protease activities in ground beef were examined and increased as the temperature increased. Experiments varying both the concentrations of added bacteriocin SH01 and temperature demonstrated a maximum inhibition (2.33 log reduction of bacteria) in samples containing 1,000 AU/g of bacteriocin SH01 incubated at 20℃. When the crude bacteriocin SH01 solution (1,280 AU/mL) was incubated with crude protease solutions at different temperatures, its activity decreased by only half (640 AU/mL), as assessed in an agar well diffusion assay. The finding that the antilisterial activity of bacteriocin SH01 increased with temperature can be explained by the fact that higher temperatures increase bacterial membrane fluidity, thereby promoting the cellular penetration of bacteriocin SH01 into L. monocytogenes. Bacteriocin SH01 may be an excellent candidate as a biopreservative for controlling L. monocytogenes growth in ground beef.

• Food Engineering Progress
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• v.13 no.3
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• pp.176-182
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• 2009

This study was carried out to analyse the physicochemical characteristics of soybean pastes with the sword beans and general soybean pastes. There was no difference in contents of general component (moisture contents, crude ash, crude fiber, crude fat) among the soybean pastes, but contents of crude protein was higher in J1, J2 compared to the others. Amylase activity was higher in J3, J4 adding the rice, and protease activity was higher in J1, J2 that has higher soybean contents than the others. Reducing sugar contents was higher in J3, J4 that has high amlyase activity with value of 8.53${\pm}$0.45% and 8.36${\pm}$1.05%, and amino-type nitrogen contents was higher in commercial soybean paste K5, K6 than other soybean pastes with value of 668.34${\pm}$4.11 mg% and 642.64${\pm}$2.05 mg% respectively.

• Journal of Pharmaceutical Investigation
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• v.19 no.1
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• pp.9-14
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• 1989

In order to study the protease from Agaricus bisporus (Lange), the crude protease preparation was separated by fractionation of mushroom extracts with ammonium sulfate. It was found that extracts from Agaricus bisporus (Lange) Sing. contained protease. The optimum pH of the enzyme was 6.0, and the pH range at which the enzyme was stable was 4.0 to 7.0. The optimum temperature at which the enzyme showed the highest proteolytic activity was $50^{\circ}C$, while the enzyme was instantly inactivated at about $60^{\circ}C$. The enzyme activity was inhibited by $Ag^+$, $Hg^{2+}$, $Cu^{2+}$, $Ba^{2+}$, $Fe^{3+}$, $Co^{2+}$, $Ca^{2+}$, $Pb^{2+}$, $Mg^{2+}$ and $Mn^{2+}$. The $K_m$ value was 0.32 mM with Hammarsten casein.