• Title/Summary/Keyword: protase

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Cloning, Sequencing and Expression of an Extracellular Protease Gene from Serratia marcescens RH1 in Escherichia coli

  • Lee, Seung-Hwan;Kim, Jeong-Min;Kwon, Young-Tae;Kho, Young-Hee;Rho, Hyune-Mo
    • Korean Journal of Microbiology
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    • v.30 no.6
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    • pp.507-513
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    • 1992
  • Serratia marecescens RH1 isolated from soil samples produced large amount of extracellular proteases. One of the genes encoding an extracellular protease form S. marcescens RH1 was cloned in Escherichia coli by shot gun cloning method. The cloned protease, SSP, was stably expressed by its own promoter and excreted into the extracellular medium from E. coli host (ORF) of 3.135 nucleotides corresponding to 1.045 amino acids (112 kDa). The nucleotide and deduced amino acid sequence of SSP showed high overall homology (88%) to one of the S. marcescens protease (27), but low homology to other serine protease families. The optimal pH and temperature of the enzyme were pH 9.0 and 45.deg.C respectively. The activity of protease was inhibited by phenylmethylsulfonyl fluoride (PMSF), which suggests that the enzyme is a serine protease.

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Studies on the Enzyme Activities of Rhizopus distributed in South Korea(1) - On the amylase, protease and cellulase activities- (한국산 Rhizopus의 효소활성에 관한 연구 (第 1 報) - Amylase, protease 및 cellulase 활성에 관하여-)

  • 이영녹;윤경하;이평우;배광승;박용근;정성균;서항원
    • Korean Journal of Microbiology
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    • v.14 no.2
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    • pp.49-49
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    • 1976
  • Enzyme activities, such as glucoamylase dextrinogenic amylase, cellulase, acid protase and neutral protease, of Rhizopus isolated from various substrates collected throughout South Korea are measured, and their enzyme activities are surveyed from taxonomical, ecological and physiological viewpoint. Effect of carbon sources and phytohormones on the amylalse production of Rhizopus are also measured. Among the 735 strains of Phizopus isolated, strain number 587 exhibiting most prominent dextrinogenic amylase and netral protease activity is selected as the best strain, and the strain number 673, 108, 329, 165 and 728 are seleted for their predominant cellulase, acid protease, glucoamylase, dextrinogenic amylase and neutral protease activities, respectively. R.acidus and R.nigricans which exhibited relatively higher callulalse activity, showed lower activities for both amylase. R.tritici exhibited higher protease activity. The relations between activities and various substrates of wild strains are not outstnading difference, although the strains isolated from inland region exhibited more or less higher amylase and cellulase activities, than those of coast region, generally. Lactose and dextrin are most effective carbon sources for glucoamylase and dextrinogenic amylase production of the Rhizopus niveus, respectively. Although all phytohormones tested are effective for production of amylase by the Rhizopus strains, except nicotinamide for glucoamylase production, biotin and ascorbate are most effective for dextrinogenic amylase and glucoamylase production, respectively.

Studies on the Enzyme Activities of Rhizopus distributed in South Korea(1) - On the amylase, protease and cellulase activities- (한국산 Rhizopus의 효소활성에 관한 연구 (第 1 報) - Amylase, protease 및 cellulase 활성에 관하여-)

  • 이영녹;윤경하;이평우;배광승;박용근;정성균;서항원
    • Korean Journal of Microbiology
    • /
    • v.14 no.2
    • /
    • pp.47-56
    • /
    • 1976
  • Enzyme activities, such as glucoamylase dextrinogenic amylase, cellulase, acid protase and neutral protease, of Rhizopus isolated from various substrates collected throughout South Korea are measured, and their enzyme activities are surveyed from taxonomical, ecological and physiological viewpoint. Effect of carbon sources and phytohormones on the amylalse production of Rhizopus are also measured. Among the 735 strains of Phizopus isolated, strain number 587 exhibiting most prominent dextrinogenic amylase and netral protease activity is selected as the best strain, and the strain number 673, 108, 329, 165 and 728 are seleted for their predominant cellulase, acid protease, glucoamylase, dextrinogenic amylase and neutral protease activities, respectively. R.acidus and R.nigricans which exhibited relatively higher callulalse activity, showed lower activities for both amylase. R.tritici exhibited higher protease activity. The relations between activities and various substrates of wild strains are not outstnading difference, although the strains isolated from inland region exhibited more or less higher amylase and cellulase activities, than those of coast region, generally. Lactose and dextrin are most effective carbon sources for glucoamylase and dextrinogenic amylase production of the Rhizopus niveus, respectively. Although all phytohormones tested are effective for production of amylase by the Rhizopus strains, except nicotinamide for glucoamylase production, biotin and ascorbate are most effective for dextrinogenic amylase and glucoamylase production, respectively.

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Physiological properties and transformation of alkaline-tolerant bacteria (알카리내성 세균의 생리적 특성 및 형질전환)

  • 유주현;정용준;정건섭;오두환
    • Microbiology and Biotechnology Letters
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    • v.14 no.3
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    • pp.239-244
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    • 1986
  • To develop the potential use as new host strain for gene cloning, alkaline-tolerant isolates from soil were examined for amylase activity, protease activity, antimicrobial activity and transformability by using plasmid pUB 110. Of these strains, one was selected and identified as Bacillus sp. YA-14. in the enzymatic properties of Bacillus sp. YA-14 the optimal conditions for the reaction of amylase and protease were at pH 0.8 and pH 7.5 respectively. The antimicrobial activity of Bacillus sp. YA-14 was also found. For the transformation, Bacillus sp. YA-14 was cultured to late logarithmic growth phase ai 37$^{\circ}C$ in modified SPI medium (pH 8.0) containing 0.4% MgSO$_4$. The presence of pUB 110 plasmid DNA in transformants was confirmed by electrophoresis and stably maintained in the new host.

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