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[Cloning of Enterobacter aerogenes fh1A gene and overexpression of hydrogen production].
- Source :
-
Wei sheng wu xue bao = Acta microbiologica Sinica [Wei Sheng Wu Xue Bao] 2010 Jun; Vol. 50 (6), pp. 736-42. - Publication Year :
- 2010
-
Abstract
- Objective: We amplified and overexpressed the FHL activator (fh1A) in E. aerogenes ATCC13408 to enhance hydrogen production.<br />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.<br />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.
- Subjects :
- Amino Acid Sequence
Bacterial Proteins chemistry
Bacterial Proteins metabolism
Enterobacter aerogenes chemistry
Molecular Sequence Data
Sequence Alignment
Bacterial Proteins genetics
Cloning, Molecular
Enterobacter aerogenes genetics
Enterobacter aerogenes metabolism
Gene Expression
Hydrogen metabolism
Subjects
Details
- Language :
- Chinese
- ISSN :
- 0001-6209
- Volume :
- 50
- Issue :
- 6
- Database :
- MEDLINE
- Journal :
- Wei sheng wu xue bao = Acta microbiologica Sinica
- Publication Type :
- Academic Journal
- Accession number :
- 20687337