Ji Fan, Huang Danyang, Tan Xin, Guo Yong, Wang Zhen, Zhou Qing, Wang Xiaoyuan
State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, China.
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi, China.
Biotechnol Appl Biochem. 2023 Apr;70(2):716-729. doi: 10.1002/bab.2393. Epub 2022 Aug 13.
Four secondary acyltransferases of Vibrio parahaemolyticus lipid A encoded by VP_RS00880, VP_RS08405, VP_RS12170, and VP_RS01045 have been identified. In this study, mutants of V. parahaemolyticus were constructed by deleting two, three, or four of these genes. The double mutants showed similar growth pattern with the wild-type, but the quadruple mutant VPW011 showed significant growth defect at both 37°C and 21°C. Lipid A samples were extracted from these mutants and analyzed by electrospray ionization-mass spectrometry. The double and triple mutants could synthesize hepta- and octa-acylated lipid A species, while the quadruple mutant VPW011 could synthesize hexa- and hepta-acylated lipid A. The results suggest that the four secondary acyltransferases could complement each other in V. parahaemolyticus. More importantly, additional secondary acyltransferases of lipid A might exist in V. parahaemolyticus and their activities might be as strong as the four known secondary acyltransferases. The unusual multiple secondary acyltransferases of lipid A might play roles in pathogenicity and antimicrobic resistance of V. parahaemolyticus.
已鉴定出由VP_RS00880、VP_RS08405、VP_RS12170和VP_RS01045编码的副溶血性弧菌脂多糖A的四种二级酰基转移酶。在本研究中，通过缺失其中两个、三个或四个基因构建了副溶血性弧菌突变体。双突变体与野生型表现出相似的生长模式，但四突变体VPW011在37°C和21°C时均表现出明显的生长缺陷。从这些突变体中提取脂多糖A样本，并通过电喷雾电离质谱法进行分析。双突变体和三突变体能够合成七酰化和八酰化的脂多糖A种类，而四突变体VPW011能够合成六酰化和七酰化的脂多糖A。结果表明，这四种二级酰基转移酶在副溶血性弧菌中可以相互补充。更重要的是，副溶血性弧菌中可能存在其他脂多糖A的二级酰基转移酶，其活性可能与四种已知的二级酰基转移酶一样强。脂多糖A这种不寻常的多种二级酰基转移酶可能在副溶血性弧菌的致病性和抗微生物耐药性中发挥作用。
