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Biochemical characterization of 2, 3‐butanediol dehydrogenase from Taiwanofungus camphorata (583.2)
Author(s) -
Wen Lisa,
Tsai WeiWei,
Huang JenqKuen,
Lin ChiTsai
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.583.2
Subject(s) - biochemistry , dehydrogenase , enzyme , biology , formaldehyde dehydrogenase , acetoin , recombinant dna , microbiology and biotechnology , chemistry , nad+ kinase , fermentation , gene
The enzyme 2,3‐Butanediol dehydrogenase (BDH) plays important roles in reduction of acetoin to 2,3‐butanediol which is an important platform chemical with many industrial applications. Here, a TcBDH cDNA (1348 bp) 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 Ni 2+ ‐nitrilotriacetic acid Sepharose. The purified enzyme showed a single band of ~50 kD on 12% sodium dodecyl sulfate‐polyacrylamide gel electrophoresis. The Michaelis constant (K M ) value of the recombinant enzyme for acetoin was 8.46 mM. The enzyme’s optimal pH was 6. Grant Funding Source : National Science Council of the Republic of China, Taiwan under grant NSC 100‐2313‐B‐019‐003‐MY3