Earticle

초록

영어

Benzoate-CoA ligase (BCL) from Burkholderia xenovorans LB400 has narrow substrate specificity toward benzoate. Compared to the activity with benzoate, specific activity of BCL is much less than that with m-/p-hyroxybenzoate. In order to find BCL mutants which have higher activity, site-directed mutagenesis within substrate binding pocket based on rational design were carried out. The comparative structural analysis revealed that active site of 2,3-dihydroxybenzoate AMP ligase(DhbE) and 4-chlorobenzoate CoA ligase(4-CBAL) with that of BCL has shown absolutely different amino acids. Especially, substratebinding pocket of BCL showed relatively narrow circumstanse than that of BCL from Bulkholderia xenovorans due to the surrounding residues(i. e, HisXXX) located at the bottom of the active sites. Their structural analysis allows us to generate thirteen mutants and in-vitro analysis of specific activities using indirect assay monitoring NADH consumption were examined for understanding their structure-activity relationship. In conclusion, BCL mutants screened by above mentioned structural understanding and mutagenesis showed higher activity against benzoate as we are targeting.

키워드

저자

• Wonji CHOI [ School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea. ]
• Dahye JUNG [ School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea. ]
• Kwon-Young CHOI [ School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea. ]
• Eunok JUNG [ School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea. ]
• Byeori AHN [ School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea. ]
• Byung-Gee KIM [ School of Chemical and Biological Engineering, Institute of Molecular Biology and Genetics, Seoul National University, Seoul, South Korea. ]

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간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576