• Title/Summary/Keyword: protease production

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Effects of Carbon Source on Production of Leucocin A from Transformed Saccharomyces cerevisiae (형질 전환된 Saccharomyces cerevisiae의 leucocin A항균 활성도에 대한 탄소원의 영향)

  • Lee Sung-ll;Park Jin-Yong;Jung Jong-Ceun;Lee Dong-Ceun;Lee Sang-Hyeon;Ha long-Myung;Ha Bae-Jin;Lee Jae-Hwa
    • Journal of Life Science
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    • v.15 no.6 s.73
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    • pp.847-850
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    • 2005
  • The aim of this study was to increase production of leucocin A, a kind of bacteriocin, in a transformed variety of S. cerevisiae. We investigated optical density, total secreted protein, protease activity, and antibacterial activity for the transformed S. cerevisiae in different carbon sources. The production of leucocin A growth-associated, and antibacterial activity, according to carbon source, was in the order of sucrose, glucose, glycerol, and fructose. Antibacterial activity was $10.6\%$ higher in the presence of sucrose than glucose. This is the first report regarding the effect of carbon sources on the production of leucocin A in transformed S. cerevisiae, as far as we ascertain. Our results could prove useful in the industrial production of natural preservatives.

Optimization of an Industrial Medium and Culture Conditions for Probiotic Weissella cibaria JW15 Biomass Using the Plackett-Burman Design and Response Surface Methodology

  • Yu, Hyung-Seok;Lee, Na-Kyoung;Kim, Won-Ju;Lee, Do-Un;Kim, Jong-Ha;Paik, Hyun-Dong
    • Journal of Microbiology and Biotechnology
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    • v.32 no.5
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    • pp.630-637
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    • 2022
  • The objective of this study was to optimize industrial-grade media for improving the biomass production of Weissella cibaria JW15 (JW15) using a statistical approach. Eleven variables comprising three carbon sources (glucose, fructose, and sucrose), three nitrogen sources (protease peptone, yeast extract, and soy peptone), and five mineral sources (K2HPO4, potassium citrate, ⳑ-cysteine phosphate, MgSO4, and MnSO4) were screened by using the Plackett-Burman design. Consequently, glucose, sucrose, and soy peptone were used as significant variables in response surface methodology (RSM). The composition of the optimal medium (OM) was 22.35 g/l glucose, 15.57 g/l sucrose, and 10.05 g/l soy peptone, 2.0 g/l K2HPO4, 5.0 g/l sodium acetate, 0.1 g/l MgSO4·7H2O, 0.05 g/l MnSO4·H2O, and 1.0 g/l Tween 80. The OM significantly improved the biomass production of JW15 over an established commercial medium (MRS). After fermenting OM, the dry cell weight of JW15 was 4.89 g/l, which was comparable to the predicted value (4.77 g/l), and 1.67 times higher than that of the MRS medium (3.02 g/l). Correspondingly, JW15 showed a rapid and increased production of lactic and acetic acid in the OM. To perform a scale-up validation, batch fermentation was executed in a 5-l bioreactor at 37℃ with or without a pH control at 6.0 ± 0.1. The biomass production of JW15 significantly improved (1.98 times higher) under the pH control, and the cost of OM was reduced by two-thirds compared to that in the MRS medium. In conclusion, OM may be utilized for mass producing JW15 for industrial use.

Studies on the Proteolytic Enzyme of Mold (Part 1) Production of Acid Protease by Aspergillus awamori U-3 and Characteristics of Enzyme (사상균의 단백질분해효소에 관한 연구 (제1보) Aspergillus awamori U-3에 의한 Acid Protease의 생산 및 효소의 특성)

  • 정만재;박남규
    • Microbiology and Biotechnology Letters
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    • v.7 no.3
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    • pp.157-164
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    • 1979
  • These experiments were performed to investigate the culture condition, characteristic of crude enzyme and the heat resistance of the acid protease by Aspergillus awamori U-3. The results obtained were as follows: 1. The optimum culture temperature and time on wheat bran medium and defatted rice bran medium were 3$0^{\circ}C$ and 72 hrs, respectively. The optimum amount of added water was 100~120 % on wheat bran medium and 100~130 % on de-fatted rice bran medium. 2. Of the these various ingredients, addition of KN $O_3$, glutamic acid and glucose on wheat bran medium and addition Of KN $O_3$, (N $H_4$)$_2$S $O_4$, glucose, lactose, K $H_2$P $O_4$ and MgC $l_2$ on defatted rice bran medium were very effective. On wheat bran medium, concentration of addition of glucose, KN $O_3$ and glutamic acid were 3.0~4.0%, 0.2~0.4 % and 1.0%, respectively. 3. The optimum pH for the enzyme action was 2.4 %, the optimum temperature about 45$^{\circ}C$ and the stable pH range 2.0~5.0, The enzyme was stable below 5$0^{\circ}C$ and was inactivated rapidly above 5$0^{\circ}C$. 4. The addition of CaC $l_2$ and CaS $O_4$ as the heat resistance agents showed the slight resistance. 5. When the enzyme solution added with the heat resistance agents (CaC1$_{2}$ and CaS $O_2$) was heated for 10-30 minutes at 6$0^{\circ}C$, their remaining activities were decreased largely above 20 minutes and The heat resistance effects of CaC $l_2$ and CaS $O_4$ were not observed almost at 8$0^{\circ}C$.

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Fermentation Characteristics of Cheese Slurry prepared from Caseinates (카세인을 이용한 치즈곤죽의 발효특성)

  • Jang, Hae-Dong;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.17 no.5
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    • pp.389-398
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    • 1985
  • To shorten the processing of cheese slurry, four different slurries, ie, Control, Cheddar 1 and 2, and Italian-type that were made of Na-caseinates, cream, trace elements, lactic culture, and enzymes were fermented at $30^{\circ}C$ for 7days with daily stirring. PH, titratable acidity, soluble nitrogen, viable cell count, active SH groups, total volatile fatty acid, free fatty acid, electrophoretic patterns of degraded caseins, and viscosity were analyzed to investigate physicochemical properties of fermented slurries. Acid production was accelerated in the cheese slurries with protease than that without the enzyme and PH of the former was decreased after three days of fermentation to 4.90. The Change of titratable acidity agreed to PH patterns. Soluble nitrogen of the Control slurry was increased slowly for four days and then rapidly to 40% of total nitrogen while those containing protease to 70%. The protease of lactic cultures used (Streptococcus lactis and Streptococcus cremoris) broke down as-casein more rapidly than $\beta$-casein and most proteins were degraded to peptides and amino acids after three days of fermentation. Total volatile fatty acids were increased by added lipase and free fatty acids composition analyzed by GLC in cheddar slurry with 0.00001% lipase was similar to that of commercial cheddar cheese, while that in Italian-type slurry was a half of that in commercial Italian cheese. Active SH groups were increased in the cheese slurries with glutathione from fourth day of fermentation. The viscosity of slurries decreased very rapidly by addition of protease.

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Evaluation of Strains of Metarhizium anisopliae and Beauveria bassiana against Spodoptera litura on the Basis of Their Virulence, Germination Rate, Conidia Production, Radial Growth and Enzyme Activity

  • Petlamul, Wanida;Prasertsan, Poonsuk
    • Mycobiology
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    • v.40 no.2
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    • pp.111-116
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    • 2012
  • Ten strains of the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana were evaluated to find the most effective strain for optimization studies. The first criterion tested for strain selection was the mortality (> 50%) of Spodoptera litura larvae after inoculation of the fungus for 4 days. Results on several bioassays revealed that B. bassiana BNBCRC showed the most virulence on mortality S. litura larvae (80% mortality). B. bassiana BNBCRC also showed the highest germination rate (72.22%). However, its conidia yield ($7.2{\times}10^8$ conidia/mL) was lower than those of B. bassiana B 14841 ($8.3{\times}10^8$ conidia/mL) and M. anisopliae M6 ($8.2{\times}10^8$ conidia/mL). The highest accumulative radial growth was obtained from the strain B14841 (37.10 mm/day) while the strain BNBCRC showed moderate radial growth (24.40 mm/day). M. anisopliae M6 possessed the highest protease activity (145.00 mU/mL) while M. anisopliae M8 possessed the highest chitinase activity (20.00 mU/mL) during 96~144 hr cultivation. Amongst these criteria, selection based on virulence and germination rate lead to the selection of B. bassiana BNBCRC. B. bassiana B14841 would be selected if based on growth rate while M. anisopliae M6 and M8 possessed the highest enzyme activities.

Isolation and Characterization of Transposon \ulcorner¨ªKm-Mediated Nonpathogenic Mutants of Xanthomonas campestris pv. vesicatoria (고추 세균성 반점병균의 비병원성 돌연변이체 분리 및 생리적 특성)

  • 윤영채;김용식;조용섭
    • Korean Journal Plant Pathology
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    • v.11 no.3
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    • pp.265-270
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    • 1995
  • Transposon mutation of Xanthomonas campestris pv. vesicatoria (Xcv) was induced by using transposon omegon ($\Omega$)-Km (Tn $\Omega$Km), which was confirmed by resistance to kanamycin (KMr), and nonpathogenic mutants were selected through the inoculation test on pepper plants. The mutagenesis frequency was about 6$\times$10-8, and 53 out of 2,000 Kmr bacterial colonies tested were nonpathogenic to the pepper cultivar Cheung-Hong. Optimum conditions for the Tn $\Omega$Km mutagenesis of Xcv were Luria Bertani (LB) broth medium for culture of Xcv, yeast extract-dextrose-CaCO3 (YDC) agar medium for selection of Tn $\Omega$Km-mediated mutants, and over 1 to 2 in the ratio of the donor (Escherichia coli S17-1 with the plasmid pJFF350 $\Omega$Km) and the recipient (Xcv) in the culture for the mutagenesis. One of the 4 nonpathogenic mutants (WNP1, WNP3, WNP4 and WNP5), which had been reconfirmed through the inoculation on pepper cv. Dabokgun, showed no differences in the production of exoenzymes such as protease and polygalacturonase and extracellular polysaccharides in vitro and the bacterial growth rate from those of the wild type of Xcv.

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ORI2 is a Strong Inhibitor of Coxsackievirus B4 Replication (오리방풀로부터 분리된 ORI2의 췌장염 유발 콕사키바이러스B4 증식억제)

  • Lim, Byung-Kwan;Jo, Soyeon;Kim, Jin Hee
    • Korean Journal of Pharmacognosy
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    • v.45 no.4
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    • pp.282-287
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    • 2014
  • The ORI2 (3-[3,4-dihydroxyphenyl]acrylic acid 1-[3,4-dihydroxyphenyl]-2-methoxycarbonylethyl ester) was purified from the extract of Isodon excisus. We confirmed the antiviral effect of ORI2 in a coxsackievirus-induced pancreatitis model. Coxsackievirus B4 (CVB4) is a common cause of pancreatitis and may be reason of the type-1 diabetes. Anti-enteroviral compounds were screened by HeLa cell survival assay. Purified natural compounds were added to HeLa cells cultured 96-well plates after $10^4PFU/ml$ CVB4 pre-incubation for 30 min. ORI2 significantly improved HeLa cell survival in a dose-dependent manner. In addition, ORI2 (1 mM) treatment was dramatically decreased virus protease 2A induced eIF4G-I cleavage and viral VP1 capsid protein production. HeLa cell virus titers and viral RNA replication were significantly decreased in ORI2-treatment in a dose dependent manner (1 mM~0.001 mM). These results demonstrate that ORI2 has a strong antiviral effect. It was significantly decreased virus replication. ORI2 may be developed as a potential therapeutic agent for CVB4.

Expressed sequence tags (ESTs) analysis of Acanthamoeba healyi

  • Kong, Hyun-Hee;Hwang, Mee-Yeul;Kim, Hyo-Kyung;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.39 no.2
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    • pp.151-160
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    • 2001
  • Randomly selected 435 clones from Acanthamoeba healyi cDNA library were sequenced and a total of 387 expressed sequence tags (ESTs) had been generated. Based on the results of BLAST search, 130 clones (34.4%) were identified as the genes enconding surface Proteins , enzymes for DNA, energy Production or other metabolism, kinases and phosphatases, protease, proteins for signal transduction, structural and cytoskeletal proteins, cell cycle related proteins, transcription factors, transcription and translational machineries, and transporter proteins. Most of the genes (88.5%) are newly identified in the genus Acanthamoeba. Although 15 clones matched the genes of Acanthamoeba located in the public databases, twelve clones were actin gene which was the most frequently expressed gene in this study. These ESTs of Acanthamoeba would give valuable information to study the organism as a model system for biological investigations such as cytoskeleton or cell movement, signal transduction, transcriptional and translational regulations. These results would also provide clues to elucidate factors for pathogenesis in human granulomatous amoebic encephalitis or keratitis by Acanthamoeba.

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Production of Selenium Peptide by Autolysis of Saccharomyces cerevisiae

  • Lee Jung-Ok;Kim Young-Ok;Shin Dong-Hoon;Shin Jeong-Hyun;Kim Eun-Ki
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1041-1046
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    • 2006
  • Selenium-containing peptide (selenium peptide) was produced by autolysis of total proteins of Saccharomyces cerevisiae grown with inorganic selenium. Selenium peptide exhibited antioxidant activity as a glutathione peroxidase (GPx) mimic, and its activity was dependent on the hydrolysis methods. The GPx-like activity of the hydrolyzed selenium peptide increased 2.7-folds when digested by protease, but decreased by acid hydrolysis. During the autolysis of the yeast cell, the GPx-like activity and selenium content increased 4.3- and 2.3-folds, respectively, whereas the average molecular weight (MW) of selenium peptide decreased 70%. The GPx-like activity was dependent on the MW of selenium peptide and was the highest (220 U/mg protein) at 9,500 dalton. The maximum GPx-like activity (28,600 U/g cell) was obtained by 48 h of autolysis of the cells, which were precultured with 20 ppm of selenate. Selenium peptide showed little toxicity, compared with highly toxic inorganic selenium. These results show the potential of selenium peptide as a nontoxic antioxidant that can be produced by simple autolysis of yeast cells.

The Effects of DL-methionine and DL-methionine Hydroxy Analogue on Growth Performance, Contents of Serum Amino Acids and Activities of Digestive Proteases in Broilers

  • Lu, J.J.;Huang, C.W.;Chou, R.G.R.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.5
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    • pp.714-718
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    • 2003
  • This study was conducted to determine the effect of DL-methionine (Met) and DL-methionine hydroxy analogue (MHA) supplementation on the growth performance, contents of serum amino acids and activities of digestive protease in broiler chicks from 0 to 3 weeks old . There were three treatments in this study: (1) control (basal diet), (2) 0.24% Met supplementation and (3) 0.368% MHA supplementation. The results showed that Met and MHA supplementation did not significantly (p>0.05) improve feed efficiency and body weight gain for broilers from 0-1, 0-2 and 0-3 weeks of age. The serum levels of homocystine, methionine and taurine were significantly (p<0.05) higher with supplementation of Met or MHA than with control. The pepsin activity of proventriculus was increased with (p<0.05) Met supplementation at 21 days of age and with MHA supplementation at 7 and 14 days of age. The trypsin activity was also increased (p<0.05) with MHA supplementation at 7 days of age. The chymotrypsin activity in pancreas and the dipeptidase activity in small intestinal mucosa and content were not affected (p>0.05) by Met or MHA supplementation.