一些氨基酸对产甘油假丝酵母甘油生产的促进作用

Stimulatory Effects of Some Amino Acids on Glycerol Production by Candida glycerinogenes

以EMP途径与TCA循环中间代谢物的添加为对照,研究在尿素为氮源的产甘油假丝酵母发酵过程中添加氨基酸对甘油产量的影响。结果表明:对甘油产量有强促进作用的氨基酸有谷氨酸、谷氨酰胺、天冬氨酸、天冬酰胺、甘氨酸、赖氨酸、酪氨酸、脯氨酸、组氨酸和丝氨酸,其最适添加浓度在0.26～0.45g/L之间,丙酮酸、α_酮戊二酸、草酰乙酸、柠檬酸和琥珀酸的最适添加浓度在0.24～0.42g/L之间;赖氨酸最适于在0h添加,丙酮酸和草酰乙酸在第14h,谷氨酸、谷氨酰胺、组氨酸、脯氨酸、天冬氨酸、酪氨酸、甘氨酸、α_酮戊二酸和琥珀酸在第30h,天冬酰胺、丝氨酸和柠檬酸在第48h;在最适条件下添加这些促进剂,甘油产量均呈显著增加趋势,转化率和增加率分别达到60%和16%以上。氨基酸的作用机理为其脱氨形成的碳骨架经特定的分解代谢途径进入TCA循环,使其强化,导致碳代谢流在3_磷酸甘油醛节点处发生转移,使甘油合成途径的代谢流增加。

By using some intermediate metabolites in EMP pathway and TCA cycle as the control, the effects of amino acid supplements on glycerol production by Candida glycerinogenes in shake-flask fermentations with urea as nitrogen resource were investigated. The results showed that ten kinds of amino acids, including L-glutamic acid, L-glutamine, L-aspartic acid, L-asparagine, L-glycin, L-lysine, L-tyrosine, L-proline, L-histidine, and L-serine, had strong promotional effects on glycerol production; The optimal concentrations of these amino acids were 0.40, 0.45, 0.36, 0.35, 0.39, 0.36, 0.35, 0.45, 0.26, and 0.45 g/L, respectively. Accordingly the optimum contents of pyruvic acid, α-oxoglutarate, oxaloacetic acid, citrate, and succinate were 0.24, 0.42, 0.40, 0.37, and 0.38 g/L, respectively. The advantageous opportunities of supplement were as follows： L-lysine at the beginning of fermentation; pyruvic acid and oxaloacetic acid at the fourteenth hour; L-glntamic acid, L- glutamine, L-histidine, L-proline, L-aspartic acid, L-tyrosine, L-glycin, a- oxoglutarate, and succinate at the thirtieth hour; and L-asparagine, L-serine, and citrate at the forty-eighth hour. The addition of each stimulant at the optimal conditions could significantly promote glycerol production, with the glycerol yield on initial glucose and its increased speed exceeding 60% and 16%, respectively. The possible stimulatory mechanism due to the amino acid supplements is that the increased intermediate metabolites levels from amino acids degradation may have enhanced the flux through TCA cycle, which improve cell energetics. Meanwhile, the shift of carbon metabolism flux at the glyceraldhyde-3-phosphate node can result in the incremental flux through glycerol biosynthesis pathway.