Abstract
2,3-Butanediol dehydrogenase (Bdh) plays important roles in reduction of acetoin to 2,3-butanediol, an important platform chemical with many industrial applications. Here, a TcBdh cDNA (1348 bp, GenBank accession JF896462) encoding a putative Bdh was cloned from Taiwanofungus camphorata. The deduced amino acid sequence is similar to the Bdhs from other species. A 3-D structural model of TcBdh has been constructed based on the known structure of Pseudomonas putida formaldehyde dehydrogenase (PpFdh, PDB code 1KOL). To characterize the TcBdh protein, the coding region was subcloned into an expression vector pYEX-S1 and transformed into Saccharomyces cerevisiae. The recombinant His6-tagged TcBdh was expressed and purified by Ni2+-nitrilotriacetic acid Sepharose. The purified enzyme showed a single band of 49 kDa on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Michaelis constant (KM) value of the recombinant enzyme for acetoin was 8.5 mM. The enzyme's optical pH was 6. The thermal inactivation of the enzyme showed a half-life of 5.3 min at 45°C.
| Original language | English |
|---|---|
| Pages (from-to) | 443-448 |
| Number of pages | 6 |
| Journal | Journal of the Chinese Chemical Society |
| Volume | 62 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2015 May 1 |
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All Science Journal Classification (ASJC) codes
- Chemistry(all)
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Properties of a 2,3-butanediol dehydrogenase from taiwanofungus camphorata. / Ken, Chuian-Fu; Tsai, Wei Wei; Wen, Lisa; Sheu, Dey Chyi; Lin, Chi Tsai.
In: Journal of the Chinese Chemical Society, Vol. 62, No. 5, 01.05.2015, p. 443-448.Research output: Contribution to journal › Article
TY - JOUR
T1 - Properties of a 2,3-butanediol dehydrogenase from taiwanofungus camphorata
AU - Ken, Chuian-Fu
AU - Tsai, Wei Wei
AU - Wen, Lisa
AU - Sheu, Dey Chyi
AU - Lin, Chi Tsai
PY - 2015/5/1
Y1 - 2015/5/1
N2 - 2,3-Butanediol dehydrogenase (Bdh) plays important roles in reduction of acetoin to 2,3-butanediol, an important platform chemical with many industrial applications. Here, a TcBdh cDNA (1348 bp, GenBank accession JF896462) encoding a putative Bdh was cloned from Taiwanofungus camphorata. The deduced amino acid sequence is similar to the Bdhs from other species. A 3-D structural model of TcBdh has been constructed based on the known structure of Pseudomonas putida formaldehyde dehydrogenase (PpFdh, PDB code 1KOL). To characterize the TcBdh protein, the coding region was subcloned into an expression vector pYEX-S1 and transformed into Saccharomyces cerevisiae. The recombinant His6-tagged TcBdh was expressed and purified by Ni2+-nitrilotriacetic acid Sepharose. The purified enzyme showed a single band of 49 kDa on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Michaelis constant (KM) value of the recombinant enzyme for acetoin was 8.5 mM. The enzyme's optical pH was 6. The thermal inactivation of the enzyme showed a half-life of 5.3 min at 45°C.
AB - 2,3-Butanediol dehydrogenase (Bdh) plays important roles in reduction of acetoin to 2,3-butanediol, an important platform chemical with many industrial applications. Here, a TcBdh cDNA (1348 bp, GenBank accession JF896462) encoding a putative Bdh was cloned from Taiwanofungus camphorata. The deduced amino acid sequence is similar to the Bdhs from other species. A 3-D structural model of TcBdh has been constructed based on the known structure of Pseudomonas putida formaldehyde dehydrogenase (PpFdh, PDB code 1KOL). To characterize the TcBdh protein, the coding region was subcloned into an expression vector pYEX-S1 and transformed into Saccharomyces cerevisiae. The recombinant His6-tagged TcBdh was expressed and purified by Ni2+-nitrilotriacetic acid Sepharose. The purified enzyme showed a single band of 49 kDa on 12% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The Michaelis constant (KM) value of the recombinant enzyme for acetoin was 8.5 mM. The enzyme's optical pH was 6. The thermal inactivation of the enzyme showed a half-life of 5.3 min at 45°C.
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U2 - 10.1002/jccs.201400411
DO - 10.1002/jccs.201400411
M3 - Article
AN - SCOPUS:85028202928
VL - 62
SP - 443
EP - 448
JO - Journal of the Chinese Chemical Society
JF - Journal of the Chinese Chemical Society
SN - 0009-4536
IS - 5
ER -