Production of D-Tagatose 3-Epimerase of Pseudomonas cichorii ST-24 Using Recombinant Escherichia coli
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概要
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The D-tagaose 3-epimerase (D-TE) gene of Pseudomonas cichorii ST-24 was expressed in Escherichia coli under the control of the trc promotor. The D-TE production level was highest in E. coli JM105 as a host strain and in NZC medium as a culture medium. Production of D-TE by E. coli JM105 was about 100-fold higher than tha of D-TE by P. cichorii ST-24,and the enzyme constituted 〜5% of the total protein. D-TE from E. coli JM105 was purified by polyethylene glycol precipitation, DEAE-Toyopearl 650M column chromatography, and Sephadex G-150 column choromatograohy. The overall purification procedure resulted in 16.7-fold purification with 18.2% recovery. The moelcular weight of the purified D-TE was estimated to be 33.0 kDa by sodium dodecyl sulfate polycrylamide gel electrophoresis (SDS-PAGE), and agreed with that o the D-TE from P. cichorii ST-24. D-TE produced by E. coli JM105 had generally the same enzymological properties, i.e., the optimum pH and temperature, pH and thermal stabilities, and K_m value, as that from Pseudomonas sp. ST-24 but the V_<max> value of D-TE from E. coli was 6 times highrt than that from Pseudomonas sp. ST-35. The N-terminal amino acid sequence of the recombinant D-TE agreed with that of the D-TE from P. cichori ST-24.
- 社団法人日本生物工学会の論文
- 1997-10-25
著者
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IZUMORI KEN
Department of Bioresource Science, Faculty of Agriculture, Kagawa University
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Izumori K
Rare Sugar Research Center Kagawa University
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Izumori Ken
Rare Sugar Research Center Faculty Of Agriculture Kagawa University
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Izumori Ken
松谷化学工業 開発研究所
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Ishida Y
Department Of Bioresource Science Faculty Of Agriculture Kagawa University:department Of Biotechnolo
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ISHIDA YUTAKA
Department of Food Science, Kagawa University
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Izumori Ken
Department Of Biochemistry And Food Science Faculty Of Agriculture Kanagawa University
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Izumori Ken
Department Of Bioresource Science Faculty Of Agriculture Kagawa University
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Ishida Yuzaburo
Laboratory Of Microbiology; Department Of Fisheries Faculty Of Agriculture Kyoto University
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Izumori Ken
Department Of Biochemistry And Food Science Faculty Of Agriculture And Rare Sugar Research Center Ka
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Ishida Yutaka
Department Of Biochemistry And Food Science Faculty Of Agriculture Kagawa University
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KAMIYA TAKANORI
Department of Bioresource Science, Faculty of Agriculture, Kagawa University
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Kamiya Takanori
Department Of Bioresource Science Faculty Of Agriculture Kagawa University
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