Journal
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
Volume 163, Issue 6, Pages 707-719Publisher
SPRINGER
DOI: 10.1007/s12010-010-9076-z
Keywords
Amino acids; Corynebacterium crenatum; vgb; Fermentation; Dissolved oxygen
Funding
- High-tech Research and Development Programs of China [2006AA020301, 2007AA02Z207]
- National Basic Research Program of China [2007CB707800]
- National Natural Science Foundation of China [20676053, 30970056]
- Program for New Century Excellent Talents in University [NCET-07-0380]
- Fok Ying Tung Education Foundation [121020]
- Fundamental Research Funds for the Central Universities [JUSRP31001]
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Corynebacterium crenatum SYPA 5-5 is an aerobic and industrial l-arginine producer. It was proved that the Corynebacterium glutamicum/Escherichia coli shuttle vector pJC1 could be extended in C. crenatum efficiently when using the chloramphenicol acetyltransferase gene (cat) as a reporter under the control of promoter tac. The expression system was applied to over-express the gene vgb coding Vitreoscilla hemoglobin (VHb) to further increase the dissolved oxygen in C. crenatum. As a result, the recombinant C. crenatum containing the pJC-tac-vgb plasmid expressed VHb at a level of 3.4 nmol g(-1), and the oxygen uptake rates reached 0.25 mg A (562) (-1) h(-1) which enhanced 38.8% compared to the wild-type strain. Thus, the final l-arginine concentration of the batch fermentation reached a high level of 35.9 g L-1, and the biomass was largely increased to 6.45 g L-1, which were 17.3% and 10.5% higher than those obtained by the wild-type strain, respectively. To our knowledge, this is the first report that the efficient expression system was constructed to introduce vgb gene increasing the oxygen and energy supply for l-arginine production in C. crenatum, which supplies a good strategy for the improvement of amino acid products.
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