The Key Laboratory of Industrial Biotechnology of Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, People's Republic of China.
Appl Biochem Biotechnol. 2011 Mar;163(6):707-19. doi: 10.1007/s12010-010-9076-z. Epub 2010 Sep 11.
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.
棒状杆菌 SYPA5-5 是一株需氧、生产 L-精氨酸的工业菌株。实验证明,在协同利用氯霉素乙酰转移酶基因(cat)作为报告基因,受 tac 启动子调控的情况下,谷氨酸棒杆菌/大肠杆菌穿梭载体 pJC1 可有效地在棒状杆菌中进行扩展。利用该表达系统过表达编码黏细菌血红蛋白(VHb)的基因 vgb,进一步提高了棒状杆菌中的溶解氧。结果,含有 pJC-tac-vgb 质粒的重组棒状杆菌表达 VHb 的水平达到 3.4 nmol/g,比野生型菌株提高了 38.8%,氧摄取率达到 0.25 mg A(562)(-1)h(-1)。因此,分批发酵的最终 L-精氨酸浓度达到了 35.9 g/L,生物量也大大增加到 6.45 g/L,分别比野生型菌株提高了 17.3%和 10.5%。据我们所知,这是首次构建高效表达系统,引入 vgb 基因提高棒状杆菌中 L-精氨酸生产的氧气和能量供应,为氨基酸产品的改良提供了一个良好的策略。