UNESCO Chinese Center of Marine Biotechnology, Ocean University of China, Yushan Road, No. 5, Qingdao, China.
Mar Biotechnol (NY). 2012 Jun;14(3):261-9. doi: 10.1007/s10126-011-9409-0. Epub 2011 Oct 8.
As the β-1, 3-glucanase produced by the marine-derived Williopsis saturnus WC91-2 could inhibit the activity of the killer toxin produced by the same yeast, the WsEXG1 gene encoding exo-β-1, 3-glucanase in W. saturnus WC91-2 was disrupted. The disruptant WC91-2-2 only produced a trace amount of β-1, 3-glucanase but had much higher activity of killer toxin than W. saturnus WC91-2. After the disruption of the WsEXG1 gene, the expression of the gene was significantly decreased from 100% in the cells of W. saturnus WC91-2 to 27% in the cells of the disruptant WC91-2-2 while the expression of the killer toxin gene in W. saturnus WC91-2 and the disruptant WC91-2-2 was almost the same. During 2-l fermentation, the disruptant WC91-2-2 could produce the highest amount of killer toxin (the size of the inhibition zone was 22 ± 0.7 mm) within 36 h when the cell growth reached the middle of the log phase.
海洋来源的拟内孢霉(Williopsis saturnus)WC91-2 产生的β-1,3-葡聚糖酶可以抑制同种酵母产生的杀伤毒素的活性,因此敲除了拟内孢霉(Williopsis saturnus)WC91-2 中编码外切-β-1,3-葡聚糖酶的 WsEXG1 基因。突变株 WC91-2-2 仅产生痕量的β-1,3-葡聚糖酶,但杀伤毒素的活性比拟内孢霉(Williopsis saturnus)WC91-2 高得多。在 WsEXG1 基因敲除后,基因的表达从野生型拟内孢霉(Williopsis saturnus)WC91-2 细胞中的 100%显著降低到突变株 WC91-2-2 细胞中的 27%,而杀伤毒素基因在拟内孢霉(Williopsis saturnus)WC91-2 和突变株 WC91-2-2 中的表达几乎相同。在 2-L 发酵中,当细胞生长处于对数中期时,突变株 WC91-2-2 在 36 h 内可以产生最高量的杀伤毒素(抑菌圈大小为 22 ± 0.7 mm)。