Ji Hao, Zhuge Bin, Zong Hong, Lu Xinyao, Fang Huiying, Zhuge Jian
The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, 214122, China.
Curr Microbiol. 2016 Dec;73(6):827-833. doi: 10.1007/s00284-016-1132-7. Epub 2016 Sep 12.
Candida glycerinogenes, the glycerol producer with excellent multi-stress tolerances, is considered to be a potential biotechnological host used in the production of glycerol and its derivatives under extreme fermentation conditions. In this study, to evaluate the multiple roles of mitogen-activated protein kinase CgHOG1, we constructed a gene disruption system in the diploid C. glycerinogenes to obtain CgHOG1 null mutant. Pseudohyphae generation of the CgHOG1 mutant under non-inducing condition indicated a repressor role in morphological transitions. Disruption of CgHOG1 resulted in increased sensitivities to osmotic, acetic acid, and oxidative stress but not involved in thermotolerance. In the CgHOG1 mutant, NaCl shock failed to stimulate the accumulation of intracellular glycerol and was fatal. In addition, the CgHOG1 mutant displayed a significant prolonged growth lag phase in YPD medium with no decrease in glycerol production, whereas the mutant cannot grow under hyperosmotic condition with no detectable glycerol in broth. These results suggested that CgHOG1 plays important roles in morphogenesis and multi-stress tolerance. The growth and glycerol overproduction under osmotic stress are heavily dependent on CgHOG1 kinase.
产甘油假丝酵母是一种具有出色多胁迫耐受性的甘油生产菌,被认为是在极端发酵条件下用于生产甘油及其衍生物的潜在生物技术宿主。在本研究中,为了评估丝裂原活化蛋白激酶CgHOG1的多种作用,我们在二倍体产甘油假丝酵母中构建了一个基因破坏系统,以获得CgHOG1缺失突变体。CgHOG1突变体在非诱导条件下的假菌丝生成表明其在形态转变中起抑制作用。CgHOG1的破坏导致对渗透、乙酸和氧化应激的敏感性增加,但不涉及耐热性。在CgHOG1突变体中,NaCl冲击未能刺激细胞内甘油的积累,并且是致命的。此外,CgHOG1突变体在YPD培养基中显示出明显延长的生长滞后期,甘油产量没有下降,而该突变体在高渗条件下无法生长,肉汤中没有可检测到的甘油。这些结果表明CgHOG1在形态发生和多胁迫耐受性中起重要作用。渗透胁迫下的生长和甘油过量生产严重依赖于CgHOG1激酶。