• Food Engineering Progress
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• v.15 no.2
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• pp.182-187
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• 2011

A proteolytic lactic acid bacterium was isolated from Korean traditional fermented foods. The isolate BV-26, which had a protease activity (24 U/mg-crude protein), was identified as Lactobacillus plantarum by the API 50CHL kit and 16S rDNA analysis (99.9% of homology), and named as L. plantarum BV-26. Cell growth and protease activity of L. plantarum BV-26 was determined in MRS broth using 5L jar fermentor at $30^{\circ}C$. The maximum growth of L. plantarum BV-26 was reached at 18 hr in MRS broth, while protease activity of BV-26 was detectable at 12 hr and the highest activity was obtained after 16 hr cultivation. Therefore, we expect that the proteolytic lactic acid bacteria, L. plantarum BV-26, may be used as a starter for the fermentation of animal feed. Especially, the fermentation of soybean meal with the strain can be applied for improving feed utilization.

• Journal of Life Science
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• v.11 no.1
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• pp.42-46
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• 2001

Subtilisin YaB, produced by alkalophilic Bacillus strain YaB, is an extracellular alkaline serine protease having 55% homology to subtilisin BPN'. It is synthesized as a 378-amino acid preproenzyme and secreted into the culture medium as a 265-amino acid mature protease. To examine the role of pro-sequence for the secretion of subtilisin YaB, we have studied the expression, in Bacillus subtilis, of a mutant preprosubtilisin YaB in which active site Ser214 is substituted with Cys. The use of a six protease-deficient strain, WB600, was required for its efficient production. The prosubtilisin YaB, thus produced, was indeed secreted into the culture medium and was processed to its mature form upon treatment with exogenously added active subtilisin YaB. From these results, we have concluded that the processing of pro-sequence is not essential for the secretion of the enzyme.

• Microbiology and Biotechnology Letters
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• v.7 no.4
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• pp.205-209
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• 1979

Lactobacillus acidophilus exhibited more growth and produced greater amounts of acid in the soy milk than Streptococcus thermophilus, Lactobacillus bulgaricus, and Lactobacillu helveticus examined. The supplementation of the soy milk with glucose accelerated the growth of L. acidophilus, and enhanced acid production by L. acidophilus whereas sucrose addition was without effect. The supplementation of the soy milk containing five percent glucose with a methionine accelerated the growth of L. acidophilus, and enhannced acid production by L. acidophilus. L. acidophilus showed greater population in the soy milk containing five percent glucose which was treated with 0.0008％ protease (9.40$\times$10$^{8}$ /m/) than the soy milk containing five percent glucose (2.02$\times$10$^{9}$ /ml) moreover L. acidophilus produced greater amounts of acid in the soy milk containing five percent glucose which was treated with 0.0008％ protease (1.47 ％) than in the soy milk containing five percent glucose (0.56％)

• Microbiology and Biotechnology Letters
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• v.28 no.3
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• pp.167-171
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• 2000

Analysis of Production of Thermostable Alkaline Protease using Thermoactinomyces sp. E79. Jung, Sang Won, Sung-Sik Park, Yong-Cheol Park" Tae Kwang Oh2, and Jin-Ho Seo*, Department of Food Science and Technology, Seoul National University, Suwon 441-744, Korea, 1lnterdisciplinary program [or Biochemical Engineering & Biotechnology, Seoul National Univer5it}~ Seoul 151 "7421 Koreal 2Microbial Enzyme RU, Korea Research Institute of Bioscience & Biotechnology, Po. Box 1151 Yusong, Taejon 305"6001 Korea - This research was undertaken to analyze fermentation properties of Thermoactinomyces sp. E79 for production of a thermostable alkaline protease, which is able to specifically hydrolyze defatted soybean meal (DSM) to amino acids. TIle optimum pH for cell growth and protease production was pH 6.7, Thermoactinomyces sp. E79 did not grow at pHlO Among carbon sources tested, soluble starch was the best for protease production, while glucose repressed protease production. Tryptone was found to be the best nitrogen source for cell growth and soytone was good tor protease production. Oxygen transfer rate played an important role in producing thermostable alkaline protease. Ma'<..imum values of 6.58 glL of dry cell weight and 43.0 UJmL of protease activity were obtained in a batch fermentation using a 2.5 L jar fermentor at 1.93 X 102 hr-l of volumetric oxygen transfer coeff'jcient (kLa). Addition of 200 mgIL humic acid to the growth medium resulted in 1.64 times higher protease activity and 1.77 times higher cell growth than the case without humic acid addition.

• Parasites, Hosts and Diseases
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• v.27 no.3
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• pp.161-170
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• 1989

The proteases of Toxoplasma gcndii were purified partially and characterisrd for some biochemical properties including various chromatographic patterns, major catalytic classes, and conditions to promote the activity of these enzymes. When Toxoplasma extract was incubated with 3H-casein at various pH, peak hydrolysis of casein was observed at pH 6.0 and at pH 8.5. Proteasfs working at pH 6.0 and at pH 8.5 were purified partially by conventional methods of chromatographies of DE52 anion rxchange, Sephadex G-200 gel permeation, and hydroxylapatite chromatography. Partially purified enzymes were tested by site-specific inhibitors and promotorf. The protease working at pH 6.0 was inactivated by iodoacetamide with LDso of 10-5 M and promoted by dithiothreitol, while the protease working at pH 8.5 was inhibited by phenylmethylsulfonyl fluoride with LD50 of 10-5 M and was Promoted by ATP (excess ATP beyond 2 mM inhibited the activity reversely). The protease of pH 8.5 had the activity of ATPase which might exert the energy to its action. Therefore the former was referred to as a cysteinyl acid protease and the latter, ATP-dependent neutral serine protease.

• Fisheries and Aquatic Sciences
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• v.3 no.3_4
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• pp.188-194
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• 2000

A protease purified from hepatopancreas of shrimp, Penaeus japonicus, had maximum activity at $70^{\circ}C$ and in neutral and alkaline pH ranges. Specific activity at optimum reaction condition of the protease was estimated to be approximately 12 U/mg/min. The protease was stable in neutral and alkaline pH ranges and activity was retained after heat treatment at $50^{\circ}C$ for 30 min. Apparent $K_m$ and $V_{max}$ value against casein substrate were estimated to be $0.29\%$ and $7.8see^{-1}$, respectively, and those against N-CBZ-L-tyrosine p-nitropheny1 ester (CBZ­Tyr-NE) were 0.38 mM and $2,400 see^{-1}$, respectively. The N-termina1 sequence of the protease showed high homology to the trypsin from same species and the proteases from shrimp. Myosin heavy chain (MHC) from shrimp tail meat was the most susceptible to the protease and actin/tropomyosin were degraded progressively during 4 hr incubation, but to a lesser degree than MHC.

• Microbiology and Biotechnology Letters
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• v.5 no.3
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• pp.153-158
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• 1977

These experiments were conducted to investigate the condition of the production and the heat resistance of the acid protease by Rhizopus japonicus S-62. The results obtained were as follows: 1) The optimum concentrations of sucrose, yeast, ammonium chloride and sodium phosphate monobasic added to the wheat bran medium in the acid protease production were 0.5％, 2.0％, 0.4％, and 0.4％, respectively. 2) KH$_2$PO$_4$ and NaH$_2$PO$_4$ were the most effective as the heat resistant agents. 3) When the enzyme solutions added with KH$_2$PO$_4$ and NaH$_2$PO$_4$ to the concentration of 2％ were heated for 10 min, at 50$^{\circ}C$, their residual activities were 100％, respectively. 4) The heat resistant effects of KH$_2$PO$_4$ and NaH$_2$PO$_4$ were not observed almost above 55$^{\circ}C$.

• Textile Coloration and Finishing
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• v.20 no.2
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• pp.59-65
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• 2008

The tannin acid and the enzymes have been used in order to improve the ruggedness in laundry and the absorption of dyes and pigments in the textile industry for several years. The enzyme processing on the protein fiber minimizes the damage of the entire fiber and improves the dyeability by effectively modifying only the hydrophobic surface. This study tried out the structural observation by applying the Castanea crenata sieb. et. zucc. containing abundant tannin to the hair dyeing as the natural dyeing pigment along with Protease of Rhizopus sp. The dyeability was improved as compared to the dyeing using only the synthetic tanning and iron mordant. When the depth of pigment was higher in accordance with the surface observation, the enzyme dissolution had impact on dyeing and so the keratin layer on the hair surface. Accordingly, it was found that the appropriate depth was between 0.01 and 0.03%. It was estimated that 0.1% protease would treated within 30min. Consequently, it would cause the good reaction with the functional group of tannin pigment.

• YAKHAK HOEJI
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• v.33 no.6
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• pp.339-344
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• 1989

The alkaline protease produced by Sarcodon aspratus(Berk) S. Ito. was purified from its fruit bodies. The enzyme was purified by using ammonium sulfate fractionation, tris-acryl CM-cellulose column chromtography and chromatofocusing. The protease migrated as one major band with a molecular weight of about 29,000 dalton on sodium dodecylsulfate-polyacrylamide gel electrophoresis. The amino acid sequence of the N-terminal residues(21) of the enzyme was determined by automated sequence analysis. The sequence was Val-Thr-Thr-Lys-Gln-Thr-Asn-Ala-Pro-Trp-Gly-Leu-Gly-Asn-Ile-Ser-Thr-Thr-Asn-Lys-Leu. Comparison of this sequence with the N-terminal sequence of the p-roteinase K from Tritirachium album showed high similarity, i. e. 57.8% identical residues. The protease displayed a relatively high stability in sodium dodecyl sulfate.

• Bulletin of the Korean Chemical Society
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• v.33 no.7
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• pp.2213-2218
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• 2012

Efficient synthetic routes were developed to prepare a sizable amount (4-15 grams) of the chiral epoxides 4-6 as versatile intermediates for the synthesis of aspartyl protease inhibitors of therapeutic interest such as HIV protease and ${\beta}$-secretase. Oxidative cleavage of the C(2)-C(3) double bond of L-ascorbic acid followed by functional group manipulation led to the preparation of the epoxide 10, which was opened with an azide to yield a common aziridine intermediate 12. Through opening of the aziridine ring of 12 with either a carbon or a sulfur nucleophile, chiral epoxide precursors 4-6 could be prepared for various HIV protease inhibitors. Except for the final low melting epoxides 5 and 6, all intermediates were obtained as crystalline solids, thus the synthetic pathway can be easily applied to a large-scale synthesis of the chiral epoxides.