Earticle

초록

영어

Chromohalobacter sp. strain HS-2 was isolated from salted fermented clams and analyzed for the ability to grow on benzoate, m- and p-hydroxybenzoate derivatives as the carbon and energy source. Our previous study found that the genes responsible for the (hydroxy)benzoate degradation of HS-2 were clustered within approx. 39 kb region, which is a distinctive organization that has not been identified from those of other benzoate catabolic genes. As an initial study of the benzoate pathways, m-hydroxybenzoate hydroxylase gene (mobA) and p-hydroxybenzoate hydroxylase gene (pobA) in the cluster were determined to be overexpressed in E. coli. The two genes were amplified from HS-2 genomic DNA, cloned into an expression vector, pET-28a(+), separately and then the
cloned vectors were transformed into E. coli BL21 (DE3). The overexpression of mobA and pobA in E. coli, however, yielded significant amount of aggregated insoluble recombinant proteins. To increase solubility of recombinant MobA and PobA, the hydroxylase genes
were co-expressed with heat-shock proteins (DnaJ-DnaK). The expressed MobA and PobA were analyzed by SDS-PAGE and specific bands with a molecular weight of about 72 kDa and 44 kDa were appeared, respectively. HPLC analysis revealed that the resting cells of E.
coli BL21 (DE3) harboring mobA and pobA converted 50 mM of m- and p-hydroxybenzoate to 15 and 35 mM of protocatechuate, respectively, at 30℃.

키워드

저자

• Yu Ri PARK [ Department of Environmental Engineering, BK21 Team for Biohydrogen Production, Chosun University, Gwangju 501-759. ]
• Wonduck KIM [ Pioneer Research Center for Controlling of Harmful Algal Bloom, Chosun University, Gwangju 501-759. ]
• Si Wouk KIM [ Department of Environmental Engineering, BK21 Team for Biohydrogen Production, Chosun University, Gwangju 501-759. ]

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간행물

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• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576