Purification and Characterization of a Thermostable Alkaline Protease from Thermoactinomyces sp. E79 and the DNA Sequence of the Encoding Gene
スポンサーリンク
概要
- 論文の詳細を見る
A thermophilic Thermoactinomyces sp. E79 producing a highly thermostable alkaline protease was isolated from soil. The protease, produced extracellularly by Thermoactinomyces sp. E79, was purified by DEAE-Sepharose CL-6B and Butyl-Toyopearl 650M column chromatography. The relative molecular mass was estimated to be 31,000 by SDS-polyacrylamide gel electrophoresis. Enzyme activity was inhibited by phenylmethylsulfonyl fluoride, suggesting the enzyme to be a serine protease. The optimum temperature for the enzyme activity was 85℃, and about 50% of the original activity remained after incubation at 90℃ for 10 min in the presence of Ca^<2+>. The optimum pH for the enzyme activity was 11.0 and the enzyme was fairly stable from pH 5.0 to 12.0. The gene for this thermostable alkaline protease was cloned in Escherichia coli and the expressed intracellular enzyme was activated by heat treatment. Sequence analysis showed an open reading frame of 1,152 base pairs, coding for a polypeptide of 384 amino acids. The polypeptide was composed of a signal sequence (25 amino acids), a prosequence (81 amino acids), and a mature protein of 278 amino acids. The deduced amino acid sequence of the mature protease had high similarity with thermitase, a serine protease from Thermoactinomyces vulgaris, and the extent of sequence identity was 76%.
- 社団法人日本農芸化学会の論文
- 1996-05-23
著者
-
Kim Hyonok
Institute Of Applied Biochemistry University Of Tsukuba
-
Oh T‐k
Korea Res. Inst. Biosci. And Biotechnol. Daejon Kor
-
Kim H‐k
Hanseo Univ. Chungnam Kor
-
KIM Hyung-Kwoun
Applied Microbiology Research Group, Korea Research Institute of Bioscience & Biotechnology
-
OH Tae-Kwang
Applied Microbiology Research Group, Korea Research Institute of Bioscience & Biotechnology
-
LEE Jung-Kee
Applied Microbiology Research Group, Korea Research Institute of Bioscience and Biotechnology
-
KIM Young-Ok
Applied Microbiology Research Group, Korea Research Institute of Bioscience and Biotechnology
-
PARK Young-Seo
Food and Bioengineering, Division of Chemical and Food Engineering, Kyungwon University
-
Kim H‐k
Hanseo Univ. Seosan Kor
-
Park Young-seo
Food And Bioengineering Division Of Chemical And Food Engineering Kyungwon University
-
Lee J‐k
Korea Res. Inst. Biosci. And Biotechnol. Taejon Kor
-
Kim Young-ok
Microbial Enzyme Ru Korea Research Institute Of Bioscience & Biotechnology (kkibb)
-
Oh Tae-kwang
Microbial Enzyme Ru Korea Research Institute Of Bioscience & Biotechnology (kkibb)
-
Oh Tae-kwang
韓国生物科学生物工学研究所細菌酵素研究部門
-
Kim Hyung-kwoun
Applied Microbiology Laboratory Genetic Engineering Research Institute Korea Institute Of Science An
関連論文
- Screening for Milk-clotting Enzyme from Basidiomycetes
- Gene Cloning and Characterization of Thermostable Lipase from Bacillus stearothermophilus L1
- Heterodimeric Aminopeptidase A from Bacillus lichenformis NS115
- Partial Interfacial Activation of Proteus vulgaris Lipase Overexpressed in Escherichia coli
- Purification and Characterization of a Thermostable Alkaline Protease from Thermoactinomyces sp. E79 and the DNA Sequence of the Encoding Gene
- Antibacterial Substance Produced by Streptococcus faecium under Anaerobic Culture
- Occurrence of Thermostable Lipase in Thermophilic Bacillus Sp. Strain 398
- High-level Expression of a Recombinant Thermostable Phytase in Bacillus subtilis
- Cloning and Expression of the ccpA Gene Encoding Catabolite Control Protein from Thermoactinomyces sp. E79
- Thermostable Lipase of Bacillus stearothermophilus : High-level Production, Purification, and Calcium-dependent Thermostability
- Thermostable Chitosanase from Bacillus sp.Strain CK4 : Its Purification, Characterization, and Reaction Patterns
- Differential Response of Obese Gene Expression from Fasting in Bovine Adipose Tissues
- Induction of Osteopontin Gene Expression during Mammary Gland Involution and Effects of Glucocorticoid on its Expression in Mammary Epithelial Cells
- Sequence of a cDNA Encoding Mouse F_1F_0-ATP Synthase g Subunit
- 2種のnorfloxacin化合物をウサギに経口投与したときの体内動態の比較(短報)