title: Characterization of a thermally stable and organic solvent-adaptative NAD+ -dependent formate dehydrogenase from Bacillus sp. F1.
abstract: OBJECTIVE
  To characterize a robust NAD(+) -dependent formate dehydrogenase firstly obtained from a nonmethylotroph, Bacillus sp. F1.
  
  
  RESULTS
  The Bacillus sp. F1 NAD(+) -dependent formate dehydrogenase (BacFDH) gene was cloned by TAIL-PCR and heterologous expressed in Escherichia coli. BacFDH was stable at temperatures below 55°C, and the half-life at 60°C was determined as 52·9 min. This enzyme also showed a broad pH stability and retained more than 80% of the activities after incubating in buffers with different pH ranging from 4·5 to 10·5 for 1 h. The activity of BacFDH was significantly enhanced by some metal ions. Moreover, BacFDH exhibited high tolerance to 20% dimethyl sulfoxide, 60% acetone, 10% methanol, 20% ethanol, 60% isopropanol and 20% n-hexane. Like other FDHs, BacFDH displayed strict substrate specificity for formate.
  
  
  CONCLUSIONS
  We isolated a robust formate dehydrogenase, designated as BacFDH, which showed excellent thermal stability, organic solvent stability and a broad pH stability.
  
  
  CONCLUSIONS
  The multi-aspect stability makes BacFDH a competitive candidate for coenzyme regeneration in practical applications of chiral chemicals and pharmaceuticals synthesis with a relatively low cost, especially for the catalysis performed in extreme pH conditions and organic solvents.
authors.affiliation: Institute of Microbiology, College of Life Science, Zhejiang University, Hangzhou, China.
authors.forename: H-T
authors.initials: HT
authors.lastname: Ding
authors.forename: D-F
authors.initials: DF
authors.lastname: Liu
authors.forename: Z-L
authors.initials: ZL
authors.lastname: Li
authors.forename: Y-Q
authors.initials: YQ
authors.lastname: Du
authors.forename: X-H
authors.initials: XH
authors.lastname: Xu
authors.forename: Y-H
authors.initials: YH
authors.lastname: Zhao
mesh_terms: D000595:Amino Acid Sequence; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics; D004795:Enzyme Stability; D004926:Escherichia coli / Q000378:metabolism; D005560:Formate Dehydrogenases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006207:Half-Life; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017422:Sequence Analysis, DNA; D012997:Solvents / Q000737:chemistry*; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D012997:Solvents; D005560:Formate Dehydrogenases