title: [Cloning, expression and characterization of alcohol dehydrogenase and aldehyde dehydrogenase from Bacillus pseudofirmus OF4].
abstract: OBJECTIVE
  To clone and characterize the alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) from Bacillus pseudofirmus OF4.
  
  
  METHODS
  Genes of adh and aldh were cloned by PCR; expression vectors pET-Ahd and pET-Aldh were constructed and expressed in Escherichia coli BL21 (DE3). After Ni-NTA column chromatography purification, the protein was characterized.
  
  
  RESULTS
  The optimal temperature and pH of ALDH was 35 degrees C and 8.0, the specific activities of ALDH was 979.6 U/mg protein, the thermostability at 25 degrees C and 35 degrees C was better than at 45 degrees C. Although the expression level of ADH was too low to purify, but it was found that ADH had high catalytic activities by experiments of co-expression and ethanol tolerance.
  
  
  CONCLUSIONS
  Adh and aldh from B. pseudofirmus OF4 were cloned successfully. Co-expression of double genes could greatly increase the host strain on ethanol tolerance.
authors.affiliation: College of Life Science, Hebei Normal University, Shijiazhuang 050024, China. jujiansong@126.com
authors.forename: Jiansong
authors.initials: J
authors.lastname: Ju
authors.forename: Ning
authors.initials: N
authors.lastname: Ma
authors.forename: Ranran
authors.initials: R
authors.lastname: Zhao
authors.forename: Jingwei
authors.initials: J
authors.lastname: Liu
authors.forename: Shujing
authors.initials: S
authors.lastname: Xu
authors.forename: Baohua
authors.initials: B
authors.lastname: Zhao
mesh_terms: D000426:Alcohol Dehydrogenase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D000444:Aldehyde Dehydrogenase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D001407:Bacillus / Q000201:enzymology* / Q000235:genetics*; D003001:Cloning, Molecular; D000431:Ethanol / Q000378:metabolism
chemical_list: D000431:Ethanol; D000426:Alcohol Dehydrogenase; D000444:Aldehyde Dehydrogenase