• Title/Summary/Keyword: protease production

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Milk-clotting Enzyme from Microorganisms, Part 10, Studies on General Properties and storage of Mucor-rennin (Milk-clotting Enzyme) isolated from Mucor pusillus var. Lindt (미생물(微生物)이 생산(生産)하는 응유효소(凝乳酵素) (제10보(第10報)) -Mucor-pusillus의 고체배양(固體培養)으로부터 단리(單離)된 결정(結晶) 응유효소(凝乳酵素) Mucor-rennin의 일반적(一般的) 성질(性質)과 그의 저장성(貯藏性)-)

  • Yu, Ju-Hyun;Tamura, Gakuzo;Hong, Yun-Myung;Arima, Kei
    • 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.

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Screening and Characteristics of a Mutant of Actinoplanes teichomyceticus ATCC31121 Highly Producing Teicoplanin (Teicoplanin 생산성이 우수한 Actinoplanes teichomyceticus ATCC31121 변이주 선별 및 배양학적 특성)

  • 노용택
    • Korean Journal of Microbiology
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    • v.37 no.4
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    • pp.299-304
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    • 2001
  • Teicoplanin is a kind of glycopeptide antibiotics produced by Actinoplanes teichomyceticus, and used in the clinical antibiotic such as vancomycin against methicillin-resistant Stabphylococcus aureus (MRSA). Actino planes teichomyceticus ATCC 31121 was mutated with UV to obtain a superior mutant strain with increased level of teicoplanin production. In this investigation, lethal curve was obtained and the optimal condition to induce mutagenesis was determined to isolate the desirable mutant strain. It was also confirmed that teicoplanin activities by agar diffusion method was compared with the parent strain. One mutant strain, T991014-1 with the highest productivity, was finally selected, and was characterized through the various tests such as amylase activity, protease activity, halotolerance, antibiotic resistance, autotoxicity, and productivity. Ad fermentation characteristics of the mutant strain were also studied.

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Potential of proteolytic enzyme treatment for production of Korean red ginseng extract (홍삼 추출물의 제조에서 단백질 분해효소의 활용)

  • Kim, Dong Chung;Lee, Tae Jung;In, Man-Jin
    • Journal of Applied Biological Chemistry
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    • v.62 no.4
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    • pp.385-389
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    • 2019
  • In this study, proteolytic enzymatic treatment conditions for Korean red ginseng were examined to increase the extraction yield. Commercially available proteases were screened to obtain high protein and carbohydrate yield. The optimal dosage and reaction time for Alcalase, the chosen protease, were found to be 2.0% (w/w) and 1.5 h, respectively. Treatment with optimal conditions of Alcalase increased solid yield, total phenolic content and gensenosides content by 57.6, 81.8, and 33.8%, respectively, over levels in non-treated Korean red ginseng. Antioxidative activities evaluated by free radical scavenging activity, cation radical scavenging activity and reducing power were exactly similar between Alcalase-treated and non-treated extracts.

Characterization of Bacteriocin Produced by Enterococcus faecium MJ-14 Isolated from Meju

  • Lim, Sung-Mee;Park, Mi-Yeon;Chang, Dong-Suck
    • Food Science and Biotechnology
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    • v.14 no.1
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    • pp.49-57
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    • 2005
  • Enterococcus faecium MJ-14, having strong antilisterial activity, was isolated from Korean fermented food, Meju. MJ-14 showed the same phenotypic characteristics, but different sugar utilization, as reference strain, E. faecium KCCM12118. It could utilize D-xylose, amygdaline, and gluconate, whereas E. faecium KCCM12118 could not. Optimal condition for bacteriocin production by E. faecium MJ-14 was at $37^{\circ}C$ and pH 7.0. Bacteriocin activity appeared in mid exponential phase and increased rapidly up to stationary phase. Activity was significantly promoted in MRS broth containing 3.0% glucose, 1.5% lactose, 2.0% peptone, or 1.5% tryptone. Bacteriocins effectively inhibited Enterococcus faecalis and Listeria spp. of Gram-positive bacteria, and Helicobacter pylori of Gram-negative bacteria, but did not inhibit yeasts and molds. They were stable against heat (for 30 min at $100^{\circ}C$), pH (3.0-9.0), long-term storage (for 60 days at 4 or $-20^{\circ}C$), and enzymatic digestion by catalase, proteinase K, papain, lysozyme, trypsin, chymotrypsin, and lipase, etc. Bacteriocin activity was completely inhibited by protease and pepsin, and 50% by ${\alpha}$-amylase. Studies on PCR detection of enterocin structural genes revealed bacteriocins are identical to enterocins A and B.

Production of Hydrolyzed Red Ginseng Residue and Its Application to Lactic Acid Bacteria Cultivation

  • Kim, Dong-Chung;In, Man-Jin
    • Journal of Ginseng Research
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    • v.34 no.4
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    • pp.321-326
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    • 2010
  • Enzymatic treatment conditions for red ginseng residue (RGR) were investigated to apply RGR as a microbial medium. Polysaccharide hydrolyase and protease were screened to obtain high solid and carbohydrate yields, and a good degree of carbohydrate hydrolysis. The optimal dosage and reaction time for Viscozyme, the chosen polysaccharide hydrolyase, were found to be 1.0% (w/w) and 3 h, respectively. Of the tested proteases, Flavourzyme, whose optimal dosage was 0.5% (w/w), was selected. Co-treatment with the optimal dosages of Flavourzyme and Viscozyme increased solid yield, carbohydrate yield, and degree of carbohydrate hydrolysis by 76%, 65%, and 1,865%, respectively, over levels in non-treated RGR. The culture characteristics of Leuconostoc mesenteroides strain KACC 91459P grown in enzymatically hydrolyzed red ginseng residue (ERGR) and RGR suspensions were compared. After cultivation for 6 h, the viable cell counts of both cell suspensions rapidly increased to $1.3{\times}10^9$ colony-forming units (CFU)/g. Moreover, while the viable cell population drastically decreased to $2.4{\times}10^6\;CFU/g$ for cells grown in RGR medium, it was maintained in cells fermented in ERGR medium for 24 h.

Studies on Exudative Epidermitis in Pigs : I. Isolation and Some Properties of Staphylococcus hyicus subsp. hyicus from Diseased and Healthy Pigs (돼지 삼출성(渗出性) 표피염(表皮炎)에 관한 연구(硏究) : I. 발증돈(發症豚) 및 건강돈(健康豚)으로부터 Staphylococcus hyicus subsp. hyicus의 분리(分離) 및 그 성장(性狀))

  • Park, Cheong-kyu;Kang, Byong-kyu
    • Korean Journal of Veterinary Research
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    • v.26 no.2
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    • pp.251-257
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    • 1986
  • Isolation of Staphylococcus hyicus subsp. hyicus from piglets suffering from exudative epidermitis and healthy pigs was performed, and some properties of the isolates were examined. Twentry-six litters of exudative epidermitis observed in the field were sucking piglets with ages ranging from 1 to 5 weeks and most (73.1%) of these cases occurred in piglets between 2 and 3 weeks of age. Staphylococcus hyicus subsp. hyicus was isolated front 192(38.9%) of 494 healthy pigs. The rate of isolation of these organism from healthy pigs was found to vary greatly among farms at an isolation rate of 33.3% to 45.5% and this organism was isolated more frequently front sucking piglets below the age of 8 weeks than adult pigs. All of the isolates originated from the diseased and healthy pigs were positive in heat-stable DNase, Tween 80 hydrolysis, gelatinase, protease, but negative in clumping factor and hemelysin on sheep blood agar. There was no difference related to origin in the production of extracellular active substances. Typical lesions of the classic exudative epidermitis were produced by inoculation of Staphylococcus hyicus subsp. hyicus isolated from the diseased and healthy pigs. The pathogenic potentials of isolates from healthy pigs were no different from that of the isolates obtained from diseased pigs.

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Characterization of the Binding Activity of Virginiae Butanolide C Binding Protein in Streptomyces virginiae (Streptomyces virginiae가 생산하는 Virginiae Butanolide C(VB-C) 결합단백질의 결합활성에 미치는 일반적 특성)

  • 김현수
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.257-262
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    • 1992
  • Virginiae butanolide (VB) is an autoregulator which triggers virginiamycin production in Strefltomyces virginiae. VB-C binding protein activity was investigated under various additives. The VB-C binding protein was almost fully observed in sotubte fraction (>90%) and the binding activity was optimum at pH 7.0. The VB-C binding activity was increased about 15% in 0.5 M KCI, whereas decreased about 60% in 20 mM $Mo^{6+}$. Chelating reagents (ethylenediarnine tetraacetic acid, ethyleneglycol bis(2-aminoethylether) tetraacetic acid, 8-hydroxyquinoline) and SH protecting reagents (rnercaptoethanol, dithiothreitol, thioglycerol) inhibited the VB-C binding activity about 30~55% and 3~20%, respectively. Serine protease inhibitor (phenyl methane sulfonyl fluoride), nucleotides (guanosine 5'-monophosphate, adenosine 3',5'-cyclic monophosphate), and phosphatases (alkaline, acid phosphatase) increased the VB-C binding activity about 17%, 6~20%, and 4- 13%, respectively.

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Recombinant production of human glucagon-like peptide-1 mutant (인간 Glucagon-like Peptide-1 변이체의 재조합 생산)

  • Kim, Sung-Gun;Park, Jong-Tae
    • Korean Journal of Agricultural Science
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    • v.41 no.3
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    • pp.237-243
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    • 2014
  • Human Glucagon like peptide-1 (GLP-1) is an incretin hormone that promotes secretion of insulin. In order to eliminate the formation of the soluble aggregate, Ala19 in GLP-1 was substituted with Thr, resulting in a GLP-1 mutant GLP-1A19T. The gene synthesis of GLP-1A19T and the fusion of 6-lysine tagged ubiquitin gene were accomplished by using the overlap extension polymerase chain reaction. The ubiquitin fused GLP-1A19T (K6UbGLP-1A19T) is expressed as form of inclusion body with little formation of the soluble aggregation in recombinant E. coli. In order to produce K6UbGLP-1A19T in large amounts, fed-batch fermentation was carried out in a pH-stat feeding strategy. Maximum dry cell weight of 87.7 g/L and 20.4% of specific K6UbGLP-1A19T content were obtained. Solid-phase refolding using a cation exchanger was carried out to renature K6UbGLP-1A19T. The refolded K6UbGLP-1A19T aggregated little and was released GLP-1A19T by on-column cleavage with ubiquitin-specific protease-1. The molecular mass of GLP-1A19T showed an accurate agreement with its theoretical molecular mass.

Production of Green Tea Extract-containing Chungkook-jang (녹차 추출물을 함유하는 청국장의 제조)

  • 인만진;김동원;김동청
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.5 no.4
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    • pp.345-349
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    • 2004
  • This study was carried out to establish the manufacturing method of green tea extract-containing chungkook-jang. One strain showed the highest protease activity was isolated, and subsequently identified as Bacillus species. The strain was designated as Bacillus sp. B1, and applied to chungkook-jang fermentation. Green tea extract(TS 0.2%) was added to chungkook-jang fermentation in the quantities of 1.25%, 2.5% and 5%. Results of bacterial growth and sensory evaluation showed that chungkook-jang manufactured from addition of 1.25% quantity of peen tea extract (TS 0.2%) was most acceptable. In investigation of volatile compounds, addition of green tea extract was effective for repression of off-odor from chungkook-jang.

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Isolation of new microorganisms which degrades the protein of a food garbage efficiently and its application (음식물 쓰레기중의 단백질을 효과적으로 분해하는 신규 미생물의 분리 및 응용)

  • Koo Kyung-Wan;Chung Yong-Hyun;Hong Sung-Hee;Oh Sang-Hoon;Kim Dong-seop;Jeon Hee-Jin
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.6 no.4
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    • pp.342-348
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    • 2005
  • In this study, novel strains showing better protein degradation activity were isolated for the production of effective compost from garbages. Well growing bacteria with clear zone on the skim milk agar media were isolated . The strain was identified as Bacillus subtilis PNV-1 through various biochemical tests, Bergy's manual of determinative bacteriology and 165 rDNA partial sequence. The extracellular protease of the strain PNV-1 has its activity at broad pH and the optimal temperature was $50^{\circ}C$. Also, the strain PNV-1 was highly tolerant to high concentration of salt, red/black pepper and mustard.

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