Metabolites Study Of Recombinant Yeast Obtained By Low Energy Ion Beam Implantation

With the development of nuclear science and technology,ion beam implantation technology has been widely used not only in the fields of material modification and tumor treatment,but also in radiation breeding and low energy ion implantation in the fields of agriculture and biology.Applications are also becoming more widespread.In order to further understand the effect of low-energy ion implantation-mediated transgenic technology on yeast metabolites,based on the previous research to obtain ion beam recombinant yeasts,this study applied targeted metabolomics and nontargeted metabolomics methods by establishing Liquid chromatography-mass spectrometry(LC-MS)and ultra-high performance liquid chromatography-quadrupoletime of flight tandem mass spectrometry(UPLC-Q-TOF / MS)and nuclear magnetic resonance(NMR)analysis techniques.The intracellular and extracellular metabolites of the ion beam recombinant yeast N6076 and its original strain Kh08 were studied.Metabolomics was used to test 144 samples of ion beam recombinant yeast and its original strains at different fermentation time points,and the UPLC-Q-Tof / MS and NMR test data volume was 168 G.The results of PLS-DA analysis of intracellular metabolites showed that the composition and main component distribution of intracellular metabolites in ion beam recombinant yeast were significantly different from their original strains.The UPLC-Q-Tof / MS test results showed that the average number of small-molecule metabolites in the ion beam recombinant yeast was 88,which was 5 more than the original strain.The mass-to-charge ratio(m / z)of smallmolecule metabolites of ion beam recombinant yeasts at 0 h and 48 h is mostly between170 and 850.The small-molecule metabolites at 96 h contain compounds with m / z greater than 900.The biosynthesis time of small molecule metabolites whose molecular weight is greater than 900 is delayed by 48 h compared with the original strain.Most of the intracellular small molecule metabolites of ion beam recombinant yeast and its original strains are high abundance lipids.The difference products are mevalonate-5-phosphate(MVAP)and cystine.LC-MS / MS analysis of mevalonate(MVA)and MVAP in the mevalonate metabolism pathway showed that there was no significant MVA content at each fermentation time point of the ion beam recombinant yeast and its original strain.The difference was that the intracellular MVAP concentration of the ion-beam recombinant yeast reached 1.32 ug / m L at 48 hours after fermentation,but the original strain did not detect MVAP at the same fermentation time.The research results of extracellular products show that the products with the largest difference in ion beam recombinant yeasts and their original strains are 3-transCaffeoyltormentic acid and 3-trans-caffeoyltormentic acid(trans-3-Feruloylcorosolic acid).Acetoacetyl-CoA was found in the differential analysis of extracellular metabolites of ion beam recombinant yeasts.NMR analysis showed that the red component of the extracellular metabolites of the ion beam recombinant yeast was a quinone.This study provides a basis for further understanding of the metabolism of smallmolecule compounds in ion beam recombinant yeast cells,and lays a foundation for further research on secondary metabolites such as quinones biosynthesis by ion beam recombinant yeasts,and also validates ion beam recombinant yeasts.The results of genomics and transcriptomics analysis of bacteria provide data support,which has important scientific significance for further understanding of the effects of low energy ion implantation mediated transgenes on yeast metabolites.