Genetic and Enzymatic Studies on Thermostable α-Amylase of Bacillus subtilis Carrying the Amylase Gene from a Thermophilic Bacterium
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概要
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A Bacillus subtilis strain, 207-SV1, producing thermostable α-amylase was constructed by transformation as reported previously using a nonhomologous DNA prepared from an unidentified and Gram negative thermophilic bacterium, Thermophile V2. In the study here, enzymatic characterization of an extracellular thermostable α-amylase was conducted in comparison with B. subtilis α-amylase. Both α-amylases were found to be equally insensitive to 5mM EDTA but differed from each other in thermostability, optimum temperature, electrophoretic mobility on polyacrylamide gel, and immunological specificity. These results suggest that the structure of the thermostable α-amylase was different from that of the B. subtilis α-amylase. The structural gene for the thermostable α-amylase was named amyV2+. PBS1 phage mediated transduction and DNA mediated transformation studies with 207-SV1 and its derivative strains suggested that the chromosomal order around amyV2+ was recA-pyrA-cysC-amyV2-metC. Since the amyV2+ locus differs from that of amyE+, DNA from B. subtilis strain B7, which has a tunicamycin-resistant mutation (tmr7) located close to amyE, was transferred into strain 207-SV1, and transformants producing both thermostable α-amylase and B. subtilis α-amylase were isolated.
- 社団法人 日本農芸化学会の論文
著者
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SHINOMIYA Shoji
Institute of Applied Microbiology, The University of Tokyo
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YAMANE Kunio
Institute of Applied Microbiology, The University of Tokyo
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OSHIMA Tairo
Mitsubishi-Kasei Institute of Life Sciences
関連論文
- The Gene Responsible for Tunicamycin Resistance, tmrB, in Bacillus subtilis(Microbilolgy & Fermentation Industry)
- Gene Amplification of the amy E-tmrB Region in Bacillus subtilis
- Amplification of an Integration Plasmid Introduced into the tmr A Amplification Unit of A Bacillus subtitis Tunicamycin-resistant Mutant
- Mutational Site in Tunicamycin-resistant Mutation tmrB8 of Bacillus subtilis as Revealed at the Nucleotide Sequence Level(Microbiology & Fermentation Industry)
- recE-Dependent Gene Amplification Induced by a Tunicamycin-resistant Mutation (tmrA7) in Bacillus subtilis(Microbiology & Fermentation Industry)
- Maturation of an NH_2-Terminallly Extended Thermostable α-Amylase in Bacillus subtilis : A Possible Mechanism Examined by in Vitro Experiments
- Enhancing Effect of Bacillus subtilis Ffh, a Homologue of the SRP54 Subunit of the Mammalian Signal Recognition Particle, on the Binding of SecA to Precursors of Secretory Proteins In Vitro
- Effects of Essential Carbohydrate/Aromatic Stacking Interaction with Tyr100 and Phe259 on Substrate Binding of Cyclodextrin Glycosyltransferase from Alkalophilic Bacillus sp. 1011
- Crystal Structure of Cyclodextrin Glucanotransferase from Alkalophilic Bacillus sp. 1011 Complexed with 1-Deoxynojirimycin at 2.0 A Resolution
- Crystal Structure of Alkalophilic Asparagine 233-Replaced Cyclodextrin Glucanotransferase Complexed with an Inhibitor, Acarbose, at 2.0 A Resolution
- Molecular Cloning and Analysis of Nucleotide Sequence of the Bacillus subtilis lysA Gene Region Using B. subtilis Phage Vectors and a Multi-copy Plasmid, pUB110(Biological Chemistry)
- Expression of the Cyclodextrin Glucanotransferase Gene of Bacillus macerans in Bacillus brevis
- Protein Traffic for Secretion and Related Machinery of Bacillus subtilis
- Cloning and Nucleotide Sequence of the Bacillus subtilis K trpB Gene Encoding Tryptophan Synthase β-Subunit
- Expression of the β-Cyclodextrin Glucanotransferase Gene of an Alkalophilic Bacillus sp. #1011 in Escherichia coli Cells and Characterization of the Synthesized Enzyme(Biological Chemistry)
- The Rapid Degradation of Mutant SecA Protein in the Bacillus subtilis secA341 (ts) Mutant Causes a Protein Translocation Defect in the Cell
- Nucleotide Sequence of the Shikimate Kinase gene (aroI) of Bacillus subtilis
- Cloning of Thermostable α-Amylase Gene Using Bacillus subtilis Phage ρll as a Vector
- Cloning and Identification of an Exogenous Thermostable a-Amylase Gene residing on the Bacillus subtilis Chromosome
- Stepwise genetic transformation of Bacillus subtilis with enhancement of productivity of .ALPHA.-amylase.
- Instability of a Chimeric Plasmid Containing Bacillus subtilis aroI+ tmrB Genes, pBR322 and pUBHO, during B. subtilis Transformation
- Genetic and Enzymatic Studies on Thermostable α-Amylase of Bacillus subtilis Carrying the Amylase Gene from a Thermophilic Bacterium
- Enhancing Effect of Bacillus subtilis Ffh, a Homologue of the SRP54 Subunit of the Mammalian Signal Recognition Particle, on the Binding of SecA to Precursors of Secretory Proteins In Vitro.