• Journal of Microbiology and Biotechnology
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• v.6 no.1
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• pp.1-6
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• 1996

A bacterial strain NS70 producing an alkaline protease was isolated from soil samples taken near a hot spring and identified as Bacillus licheniformis by its morphological and physiological properties and cellular fatty acid analysis. The isolated alkaline protease was purified by ammonium sulfate fractionation, DEAE-, CM-, and Phenyl-Sepharose column chromatography. The molecular weight of the purified enzyme was estimated to be 32, 000 Da by sodium dodecylsulfate polyacrylamide gel electrophoresis. Its optimal pH and temperature for proteolytic activity against Hammarsten casein were 12 and $65^{\circ}C$, respectively. The enzyme was stable at alkaline pH range from 6.0 to 12.0, and fairly stable up to $65^{\circ}C$. The enzyme was inhibited by phenylmethylsulfonyl fluoride but not by EDTA and N-ethylmaleimide indicating that the enzyme is serine protease. Enzyme activity was markedly inhibited by $Hg^{2+}$ and $Cu^{2+}$. Autolytic phenomena were observed on purified protease NS70 but autolysis was reduced by the addtion of $Ca^{2+}$ ion or bovine serum albumin.

• BMB Reports
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• v.44 no.6
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• pp.387-392
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• 2011

To investigate the molecular scavenging capabilities of the larvae of Hermetia illucens, two serine proteases (SPs) were cloned and characterized. Multiple sequence alignments and phylogenetic tree analysis of the deduced amino acid sequences of Hi-SP1 and Hi-SP2 were suggested that Hi-SP1 may be a chymotrypsin- and Hi-SP2 may be a trypsin-like protease. Hi-SP1 and Hi-SP2 3-D homology models revealed that a catalytic triad, three disulfide bonds, and a substrate-binding pocket were highly conserved, as would be expected of a SP. E. coli expressed Hi-SP1 and Hi-SP2 showed chymotrypsin or trypsin activities, respectively. Hi-SP2 mRNAs were consistently expressed during larval development. In contrast, the expression of Hi-SP1 mRNA fluctuated between feeding and molting stages and disappeared at the pupal stages. These expression pattern differences suggest that Hi-SP1 may be a larval specific chymotrypsin-like protease involved with food digestion, while Hi-SP2 may be a trypsin-like protease with diverse functions at different stages.

• Animal Bioscience
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• v.36 no.12
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• pp.1869-1879
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• 2023

Objective: This study was to evaluate the effects of individual or combinational use of phytase, protease, and xylanase on total tract digestibility of corn, soybean meal, and distillers dried grains with soluble (DDGS) fed to pigs. Methods: Each experiment had four 4×4 Latin squares using 16 barrows. Each period had 5-d adaptation and 3-d collection. All experiments had: CON (no enzyme); Phy (CON+phytase); Xyl (CON+xylanase); Pro (CON+protease); Phy+Xyl; Phy+Pro, Xyl+Pro, Phy+Xyl+Pro. Each Latin square had 'CON, Phy, Xyl, and Phy+Xyl'; 'CON, Phy, Pro, and Phy+Pro'; 'CON, Pro, Xyl, and Xyl+Pro'; and 'Phy+Xyl, Phy+Pro, Xyl+Pro, Phy+Xyl+Pro'. Results: The digestible energy (DE), metabolizable energy (ME), and nitrogen retention (NR) of corn were not affected by enzymes but the apparent total tract digestibility (ATTD) of phosphorus (P) was improved (p<0.01) by Phy. The DE and ATTD dry matter (DM) in soybean meal were increased (p<0.05) by Phy+Pro and the ATTD P was improved (p<0.01) by Phy, Phy+Pro, and Phy+Xyl. The DE, ME, and ATTD DM in DDGS were improved (p<0.05) by Phy+Xyl and the ATTD P was improved (p<0.01) by Phy, Phy+Pro, and Phy+Xyl. Conclusion: Phytase individually or in combination with xylanase and protease improved the Ca and P digestibility of corn, soybean meal, and DDGS, from the hydrolysis of phytic acid. The supplementation of protease was more effective when combined with phytase and xylanase in the soybean meal and DDGS possibly due to a higher protein content in these feedstuffs. Xylanase was more effective in DDGS diets due to the elevated levels of non-starch polysaccharides in these feedstuffs. However, when xylanase was combined with phytase, it demonstrated a higher efficacy improving the nutrient digestibility of pigs. Overall, combinational uses of feed enzymes can be more efficient for nutrient utilization in soybean meal and DDGS than single enzymes.

• Journal of the Korean Society of Food Science and Nutrition
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• v.27 no.6
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• pp.1077-1085
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• 1998

The effects of yeast on the fermentation of kimchi were investigated. The treatments were divided into two groups; yeast treatment during salting of Chinese cabbage(YS) and yeast treatment added in kimchi preparation(YF kimchi). The edible periods of the kimchi after yeast treatment during salting (YS kimchi) was extended 4~5 days by the results of pH, acidity, sensory quality. The activities of amylase, polygalacturonase and galactosidase of YS kimchi were retained at low levels compared to non treated condition throughout all fermentation periods, whereas protease activity was not significant different from the non treated condition. In addition, the contents of total hexose and uronic acid did not show remarkable change throughout fermentation, but total pentose was decreased by more than 7% at the early middle stage of fermentation(7~14 day after soaking). The change of free amino acid content was decreased by 16~44% than the non treated condition. In contrast, in the YF kimchi, the sensory quality was not good. The activities of amylase, protease, polygalacturonase and gal actosidase were appreciably higher than that of the non treated condition. Meanwhile, the contents of total hexose, total pentose and uronic acid, as products of degradation of cell wall constituents by the above enzymes, were decreased by 18~68% throughout fermentation than the non treated con dition, and total free amino acids were higher than the YS kimchi. Thus, yeast treatment during salting was found to be more effective to extend the edible periods of the kimchi.

• Korean Journal of Food Science and Technology
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• v.3 no.1
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• pp.64-67
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• 1971

The Manufacturing conditions of Chungkukjang were investigated by the use of the soy beans, salt (NaCl) and Bacillus subtilis S.P. In the studies of Chungkukjang Meju protease activity, total acid, free amino nitrogen, optimum times of fermentation and the maturity of Meju were shown the reasonable conditions each at 54 hrs, 24 hrs, between 36 to 42 hrs, between 36 to 42 hrs and at 5 days from the beginning of fermentation. In the studies on the Chungkukjang (Sauce made by meju) the increased amount of free amino nitrogen and total acid were proportional to the increased contents of moisture. The amount of the acid was increased rapidly for 20 day from the beginning and after it slowly. The appropriate content of moisture was 55% in the manufacturing of Chungkukjang.

• Asian-Australasian Journal of Animal Sciences
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• v.23 no.6
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• pp.700-707
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• 2010

Effects of protease-resistant antimicrobial substances (PRA) produced by Lactobacillus plantarum and Leuconostoc citreum on rumen methanogenesis were examined using the in vitro continuous methane quantification system. Four different strains of lactic acid bacteria, i) Lactococcus lactis ATCC19435 (Control, non-antibacterial substances), ii) Lactococcus lactis NCIMB702054 (Nisin-Z), iii) Lactobacillus plantarum TUA1490L (PRA-1), and iv) Leuconostoc citreum JCM9698 (PRA-2) were individually cultured in GYEKP medium. An 80 ml aliquot of each supernatant was inoculated into phosphate-buffered rumen fluid. PRA-1 remarkably decreased cumulative methane production, though propionate, butyrate and ammonia N decreased. For PRA-2, there were no effects on $CH_4$ and $CO_2$ production and fermentation characteristics in mixed rumen cultures. The results suggested that PRA-1 reduced the number of methanogens or inhibited utilization of hydrogen in rumen fermentation.

• Microbiology and Biotechnology Letters
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• v.29 no.2
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• pp.110-114
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• 2001

Isolation of BacilLus sp. producing multi-enzyme and optimization of medium conditions for its production using feedstuffs for probiotics were carried out in this study. A bacterium isolated from natural resources, namely Bacillus subtilis 4-3, has multi-enzyme activity (phytase. cellulase, xylanasc, protease, and amylase. In the culture of B. subtilis 4-3 using soybean meal and rice bran. relatively low phytate degradation was noted using whereas high phytate degradability was observed with wheat bran (80.63%). The optimal composition of medium using feedstuffs was 1.0% (w/v) soybean meal and 2% (w/v) molasses to yield high cell growth.

• Korean Journal of Food Science and Technology
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• v.12 no.1
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• pp.13-17
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• 1980

A Streptomyces spp. strain SY 79-1 which was capable of producing metal protease was isolated from soil. The optimal pH and temperature of the protease were around pH 8.0 and $45^{\circ}C$, respectively. The stable pH range of the enzyme was between pH 6.0 to 8.0. The enzyme was stable at $45^{\circ}C$, but it lost the activity about 75 % for 5 min and completely for 30 min when it was treated at $60^{\circ}C$. The activity of the enzyme was inhibited by $Hg^{++},\;Cu^{++},\;Ag^{+}$ and activated by $Mg^{++},\;Mn^{++},\;Co^{++},\;but\;Fe^{++},\;Ca^{++},\;Pb^{++}\;and\;Al^{3+}$ did not affect enzyme activity. This enzyme was strongly inhibited by EDTA, but was not inhibited by 2, 4-DNP, ${\rho}$-CMB, ${\varepsilon}$-aminocaproic acid, cysteine, thiourea, citric acid, oxalic acid and sodium arsenate. When cobalt was added to the EDTA-denatured enzyme, the activity of the enzyme was restored.

• Journal of Sericultural and Entomological Science
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• v.31 no.1
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• pp.37-44
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• 1989

The parasporal crystals of Bacillus thuringinsis subspecies kurstaki, dendrolimus, finitimus, aizawai and israelensis were compared by polyacrylamide electrophoresis, amino acid composition and immunological analysis. In the subspecies of kurstaki, dendrolimus, finitimus and aizawai, the molecular weights of main subusnits of crystal solubilized by alkaline solution were 1.3${\times}$105 and 6.5${\times}$104 while those of subsp. israelensis were 4${\times}$104 and 1,4${\times}$104. The degradation of lepidopteran toxic subspecies crystals by silkworm midgut protease showed 6.0-6.4${\times}$104 molecular weight and the pattern of degradation of subsp. israelensis crystals was similar to that of alkaline solution treatment. In the amino acid composition, aspartic acid in subsp. israelensis and glutiamic acid in the other four subspecies were the most abundant. The immunological characteristics of the crystals revealed that the antibody produced against the alkali-solubilized crystal protein of subsp. israelensis reacted with only its antigen, but the crystal antigens from the other four lepidopteran toxic subspecies did cross-react with each other as well as with their own homologous antisera.

• Journal of Life Science
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• v.20 no.7
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• pp.1005-1011
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• 2010

$\gamma$-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system. GABA transporters (GATs) control extracellular GABA levels by reuptake of released GABA from the synaptic cleft. However, how GATs are regulated has not yet been elucidated. Here, we used the yeast two-hybrid system to identify the specific binding protein(s) that interacts with the carboxyl (C)-terminal region of GAT1, the major isoform in the brain and find a specific interaction with the ubiquitin-specific protease 14 (Usp14), a deubiquitinating enzyme. Usp14 protein bound to the tail region of GAT1 and GAT2 but not to other GAT members in the yeast two-hybrid assay. The C-terminal region of Usp14 is essential for interaction with GAT1. In addition, these proteins showed specific interactions in the glutathione S-transferase (GST) pull-down assay. An antibody to GAT1 specifically co-immunoprecipitated Usp14 from mouse brain extracts. These results suggest that Usp14 may regulate the number of GAT1 at the cell surface.