The Regulation Mechanism Of Acylation Modification Of Isocitrate Lyase In M.smegmatis

Lysine acylation is a very important mechanism of protein function regulation in the living body and is widely involved in the regulation of activities such as carbon metabolism.The MsICL activity affects the distribution of carbon metabolism between the TCA cycle and the glyoxylate pathway,and affects the metabolism of fatty acids and other carbon substrates in M.smegmatis.In order to reveal the effect of acetylation/propionylation modification on MsICL1,we studied in depth the regulatory mechanisms of acetylation/propionylation modification of MsICL1 and analyzed the effect of acylation modification of MsICL1 on carbon metabolism in M.smegmatis.This article first constructed MsPat-deleted bacteria and replenishing bacteria.The knockout plasmid pPR27-Mspat was constructed by restriction-ligation reaction and PCR and transformed into competent cells of M.smegmatis by electroporation.The positive characteristics of knockout vectors were used to screen positive knockout strains and gene sequencing and PCR to verify the construction of knockout bacteria.Based on this,the effects of the lack of MsPat on the apparent morphology of the bacteria were also discussed.The deletion of MsPat had little effect on cell morphology and biofilm formation,but inhibited the ability to resist rifampicin and isoniazid and significantly reduced the degree of acetylation of the whole protein,indicating that the metabolic processes of different carbon substrates were regulated.This study not only revealed the significance of acetylation modification in M.smegmatis,but also provided a new idea for the development of a new protein acylation modification.ICL is a key metabolic enzyme in the glyoxylate cycle,which plays an important role in the distribution of carbon metabolic fluxes in the glyoxylate cycle and the TCA cycle,and is an important protease for the metabolism of fatty acids and other carbon substrates.Studies have shown that acetylation and propionylation modify MsICL1 and affect its activity.This acylation modification is catalyzed by MsPat.The loss of MsPat has an adverse effect on the metabolism of propionate,while there is no significant effect of carbon metabolism such as acetate,cholesterol and glucose.This study not only reveals the mechanism of the modification of the acylation of MsICL1,but also has implications for the analysis of the mechanism of acylation modification of mycobacterial ICL and opens up new directions for the study of antituberculosis drugs.