Amiri-Jami Mitra, Wang Haifeng, Kakuda Yukio, Griffiths Mansel W
Department of Food Science, University of Guelph, N1G 2W1, Guelph, ON, Canada.
Biotechnol Lett. 2006 Aug;28(15):1187-92. doi: 10.1007/s10529-006-9077-8. Epub 2006 Jun 24.
Transposon Tn5 mutagenesis was used to generate random mutations in Shewanella baltica MAC1, a polyunsaturated fatty acid (PUFA)-producing bacterium. Three mutants produced 3-5 times more eicosapentaenoic acid (EPA 20:5 n-3) compared to the wild type at 10 degrees C. One of the mutants produced 0.3 mg EPA g(-1) when grown at high temperature (30 degrees C). Moreover, 2 mg docosahexaenoic acid (DHA 22:6 n-3) g(-1) was produced by S. baltica mutants at 4 degrees C. Sequencing of insertion mutation(s) showed 96% homology to trimethylamine N-oxide (TMAO) reductase gene and 85% homology to rRNA operons of E. coli. Tn5 transposon mutagenesis therefore is a suitable technique to increase PUFA formation in bacteria.
转座子Tn5诱变被用于在波罗的海希瓦氏菌MAC1(一种产生多不饱和脂肪酸的细菌)中产生随机突变。在10摄氏度时,三个突变体产生的二十碳五烯酸(EPA 20:5 n-3)比野生型多3至5倍。其中一个突变体在高温(30摄氏度)下生长时产生了0.3毫克EPA g(-1)。此外,波罗的海希瓦氏菌突变体在4摄氏度时产生了2毫克二十二碳六烯酸(DHA 22:6 n-3)g(-1)。插入突变的测序显示与三甲胺N-氧化物(TMAO)还原酶基因有96%的同源性,与大肠杆菌的rRNA操纵子有85%的同源性。因此,Tn5转座子诱变是一种增加细菌中多不饱和脂肪酸形成的合适技术。