Improvement of Poly(3-Hydroxybutyrate)[P(3HB)] Production in Corynebacterium glutamicum by Codon Optimization, Point Mutation and Gene Dosage of P(3HB) Biosynthetic Genes(ENZYMOLOGY, PROTEIN ENGINEERING, AND ENZYME TECHNOLOGY)
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概要
- 論文の詳細を見る
In our previous study, a system for producing poly(3-hydroxybutyrate) [P(3HB)] was established by introducing a polyhydroxyalkanoate (PHA) biosynthetic gene operon (phaCAB_<Re>) derived from Ralstonia eutropha into Corynebacterium glutamicum. In this study, two experimental strategies have been applied to improve P(3HB) production in recombinant C. glutamicum. One is a codon optimization of the N-terminal-coding region of the PHA synthase (PhaC_<Re>) gene focusing on the codon usage preference for the translation system of C. glutamicum. The other is the replacement of wild-type phaC_<Re> with a modified gene encoding a mutation of Gly4Asp (G4D), which enhanced the production of PhaC_<Re> and P(3HB) in Escherichia coli. The introduction of these engineered PHA synthase genes into C. glutamicum enhanced the production of PhaC_<Re> and P(3HB). Interestingly, we found that these gene modifications also caused increases in the concentration of the translation products of the genes encoding monomer-supplying enzymes, β-keto-thiolase (PhaA_<Re>) and acetoacetyl-CoA reductase (PhaB_<Re>). This finding prompted us to carry out a gene dosage of phaAB_<Re> for a double plasmid system, and the highest production (52.5wt%) of P(3HB) was finally achieved by combining the gene dosage of phaAB_<Re> with codon optimization. The molecular weight of P(3HB) was also increased by approximately 2-fold, as was P(3HB) content. Microscopic observation revealed that the volume of the cells accumulating P(3HB) was increased by more than 4-fold compared with the non-P(3HB)-accumulating cells without filamentous morphologenesis observed in E. coli.
- 社団法人日本生物工学会の論文
- 2007-12-25
著者
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LEONG Chean
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido Uni
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JO Sung-Jin
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido Uni
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Leong Chean
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Matsumoto Ken'ichiro
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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TAGUCHI Seiichi
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido Uni
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Ooi Toshihiko
Division of Biotechnology and Macromolecular Chemistry, Graduate School of Engineering, Hokkaido Uni
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Jo Sung-jin
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Taguchi Seiichi
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Sun Jian
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Taguchi Seiichi
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Matsumoto Ken'ichiro
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Morimoto Kenjiro
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Ooi Toshihiko
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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Taguchi Seiichi
Division Of Biotechnology And Macromolecular Chemistry Graduate School Of Engineering Hokkaido Unive
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