| 학회 |
한국화학공학회 |
| 학술대회 |
2012년 가을 (10/24 ~ 10/26, 부산 BEXCO) |
| 권호 |
18권 2호, p.1903 |
| 발표분야 |
생물화공 |
| 제목 |
Cloning, expression and characterization of 3-hydroxyisobutyrate dehydrogenase from Pseudomonas denitrificans ATCC13867 |
| 초록 |
The gene encoding an NAD+-dependent 3-hydroxyisobutyrate dehydrogenase (3HIBDH-IV) from Pseudomonas denitrificans ATCC 13867 was cloned and expressed in Escherichia coli BL 21 (DE3) and characterized to understand its physiological relevance in 3-hydroxypropionic acid (3-HP) degradation. The deduced amino acid sequence exhibited a high similarity to other 3-hydroxyisobutyrate dehydrogenase isozymes (3HIBDHs) of P. denitrificans ATCC 13867. The comparison of 3HIBDH-IV with its relevant enzymes indicates that Lys171 is important for catalytic function on 3-hydroxyacids. Purified 3HIBDH-IV is specific to (S)-3-hydroxyisobutyrate, but also catalyzes the oxidation of 3-HP to malonate semialdehyde. The half-saturation constant (Km) for 3-HP was 1.0 mM and the specific activity of 3HIBDH-IV was 17 U/mg protein for 3-HP with NAD+ as a cofactor at 30 oC and pH 9. The specific activity constant (Kcat/Km) for the oxidation of 3-HP was estimated at 10.24×103 M-1S-1. The heavy metals such as Ag+ and Hg2+ inhibited 3HIBDH-IV activity, whereas dithiothreitol, 2-mercaptoethanol and ethylenediamine- tetraacetic acid increased its activity by 1.5–1.8 fold. |
| 저자 |
Shengfang Zhou, 김하나, 박성훈
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| 소속 |
부산대 |
| 키워드 |
3-hydroxyisobutyrate dehydrogenase; 3-hydroxyisobutyric acid; 3-hydroxypropionic acid; ; 3-hydroxypropionate degradation; Pseudomonas denitrificans
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| E-Mail |
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| 원문파일 |
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