title: [Cloning of Enterobacter aerogenes fh1A gene and overexpression of hydrogen production].
abstract: OBJECTIVE
  We amplified and overexpressed the FHL activator (fh1A) in E. aerogenes ATCC13408 to enhance hydrogen production.
  
  
  METHODS
  By using universal primers and genome walking, we cloned the full open reading frame (ORF) of fh1A gene. We inserted it into the glutathion S-transferase (GST) fusion expression vector pGEX4T-2-Cat, and transformed the recombinant plasmid into E. aerogenes ATCC13408 via electroporation for expression. Then we measured the hydrogen production of the recombinant strain in a batch culture.
  
  
  RESULTS
  We found that the ORF of fh1A was 2073 base pair in length, potential to encode a 690 amino acid peptide (GenBank accession GU188474). The Fh1A protein from E. aerogenes ATCC13408 shared high amino acid identities with those from other bacterial species. By using SDS-PAGE and Western blot analysis, we confirmed that the fh1A gene had successfully expressed in the strain. The hydrogen yield of the recombinant strain was increased from 1.23 to 1.48 mol H2/mol glucose. [ Conclusion ] Enhancement of hydrogen productivity was attained under anaerobic conditions with the recombinant strain.
authors.affiliation: Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Ministry of Agriculture, Zhejiang University, Hangzhou 310029, China). jinfangzhao@126.com
authors.forename: Jinfang
authors.initials: J
authors.lastname: Zhao
authors.forename: Wenlu
authors.initials: W
authors.lastname: Song
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.forename: Chuanxi
authors.initials: C
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D021902:Enterobacter aerogenes / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D006859:Hydrogen / Q000378:metabolism*; D008969:Molecular Sequence Data; D016415:Sequence Alignment
chemical_list: D001426:Bacterial Proteins; D006859:Hydrogen