类产碱假单胞菌XW-40发酵-生物转化级联产生D-脯氨酸  

Fermentation-biotransformation Cascade for the Production of D-proline by Pseudomonas pseudolcaligenes XW-40

作　　者：张凡凡 夏仕文[1] 谢永芳[1] 刘姣 林晖 ZHANG Fan-fan;XIA Shi-wen;XIE Yong-fang;LIU Jiao;LIN Hui(Chongqing Key Laboratory of Big Data for Bio-intelligence,College of Bioinformatics,Chongqing University of Posts and Telecommunications,Chongqing 400065,China;School of Life Sciences,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]重庆邮电大学生物信息学院,大数据生物智能重庆市重点实验室,重庆400065 [2]河南农业大学生命科学学院,河南郑州450002

出　　处：《分子催化》2021年第4期328-334,I0002,共8页Journal of Molecular Catalysis(China)

基　　金：重庆市研究生科研创新项目(CYS20270)。

摘　　要：建立了一个新颖的用于产生D-脯氨酸的发酵-生物转化过程.发酵过程中,以DL-脯氨酸为发酵前体,类产碱假单胞菌XW-40利用L-对映体诱导产生脯氨酸脱氢酶,D-对映体完全保留.在最优条件下,发酵阶段产生6 g/L D-脯氨酸.生物转化过程中,细胞不经分离,发酵液直接作为反应介质.采用分批补料策略实现DL-脯氨酸中L-对映体的转化.DL-脯氨酸单批补料浓度为10 g/L,补料次数达到5批.通过发酵和生物转化的级联,累积的D-脯氨酸浓度达到31 g/L,ee>99%.推测了生物转化过程中D-脯氨酸产生的反应机理.A novel fermentation-biotransformation cascade was developed for the production of D-proline. In the fermentation process with DL-proline as the precursor, the L-enantiomer was used to induce the production of proline dehydrogenase by Pseudomonas pseudoalcaligenes XW-40, while the D-enantiomer was completely retained. Under the optimal conditions, 6 g/L D-proline was produced in the fermentation stage. In the biotransformation process, the fermentation broth is directly used as the reaction medium without further separation of cells. The conversion of L-enantiomer in DL-proline was realized by fed-batch strategy. The feeding concentration of DLproline in a single batch was 10 g/L, and the feeding times reached 5 batches. Through the cascade of fermentation and biotransformation, the D-proline was accumulated with 31 g/L and more than 99% enantiomeric excess. The mechanism of D-proline production in biotransformation was also postulated.

关 键 词：类产碱假单胞菌XW-40 发酵 生物转化 D-脯氨酸 脯氨酸脱氢酶 

分 类 号：O643[理学—物理化学]