title: Enhancing hydrogen production of Enterobacter aerogenes by heterologous expression of hydrogenase genes originated from Synechocystis sp.
abstract: The hydrogenase genes (hoxEFUYH) of Synechocystis sp. PCC 6803 were cloned and heterologously expressed in Enterobacter aerogenes ATCC13408 for the first time in this study, and the hydrogen yield was significantly enhanced using the recombinant strain. A recombinant plasmid containing the gene in-frame with Glutathione-S-Transferase (GST) gene was transformed into E. aerogenes ATCC13408 to produce a GST-fusion protein. SDS-PAGE and western blot analysis confirm the successful expression of the hox genes. The hydrogenase activity of the recombinant strain is 237.6±9.3ml/(g-DW·h), which is 152% higher than the wild strain. The hydrogen yield of the recombinant strain is 298.3ml/g-glucose, which is 88% higher than the wild strain. During hydrogen fermentation, the recombinant strain produces more acetate and butyrate, but less ethanol. This is corresponding to the NADH metabolism in the cell due to the higher hydrogenase activity with the heterologous expression of hox genes.
authors.affiliation: State Key Laboratory of Clean Energy Utilization, Zhejiang University, Hangzhou 310027, China; Department of Life Science and Engineering, Jining University, Jining 273155, China.
authors.forename: Wenlu
authors.initials: W
authors.lastname: Song
authors.affiliation: State Key Laboratory of Clean Energy Utilization, Zhejiang University, Hangzhou 310027, China. Electronic address: juncheng@zju.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Cheng
authors.affiliation: College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, China; Key Laboratory of Fermentation Engineering (Ministry of Education), College of Bioengineering, Hubei University of Technology, Wuhan 430068, China.
authors.forename: Jinfang
authors.initials: J
authors.lastname: Zhao
authors.affiliation: College of Agriculture and Biotechnology, Zhejiang University, Hangzhou 310029, China.
authors.forename: Chuanxi
authors.initials: C
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Clean Energy Utilization, Zhejiang University, Hangzhou 310027, China.
authors.forename: Junhu
authors.initials: J
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Clean Energy Utilization, Zhejiang University, Hangzhou 310027, China.
authors.forename: Kefa
authors.initials: K
authors.lastname: Cen
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D021902:Enterobacter aerogenes / Q000235:genetics / Q000378:metabolism; D005285:Fermentation; D006859:Hydrogen / Q000032:analysis / Q000378:metabolism; D006864:Hydrogenase / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D046939:Synechocystis / Q000201:enzymology / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D011994:Recombinant Proteins; D006859:Hydrogen; D006864:Hydrogenase
keywords: Enterobacter aerogenes; Heterologous expression; HoxEFUYH; Hydrogen production; Metabolic analysis