恶臭假单胞菌TS1138转化生产L-胱氨酸的.doc

恶臭假单胞菌TS1138生产L-胱氨酸的研究(1. 南开大学生命科学学院 天津 300071(2. 天津科技大学生物工程学院 天津 300) 摘 要: 对以DL-2-氨基-?2-噻唑啉-4-羧酸DL-2-amino-?2-thiazoline-4-carboxylic acid, DL-ATC)为底物原料, 经微生物酶法催化合成L-半胱氨酸并分离L-胱氨酸恶臭假单胞菌TS1138 (Pseudomonas putida)全细胞为酶催化L-半胱氨酸以2.0%二甲基亚砜DMSO)为氧化剂氧化L-胱氨酸通过001×7型阳离子交换树脂胱氨酸高效液相色谱L-胱氨酸收率达到78.55%纯度99.12%。该方法简单高效酶稳定性差不能重复使用而固定化方法为我国L-半胱氨酸和L-胱氨酸的生产开辟一条新。恶臭假单胞菌DL-ATC, L-半胱氨酸L-胱氨酸putida TS1138 LIU Chun-Qin1 YU Yang-Sheng1 BAI Gang1* YANG Wen-Bo1CHEN Ning2 HUAI Li-Hua2 (1. College of Life Sciences, Nankai University, Tianjin 300071)(2. Bioengineering College, TianJin University of Science and Technology, Tianjin 300222) Abstract: A technology of L-cystine production was studied in this paper, which included microbial enzymatic conversion of DL-2-amino-?2-thiazoline-4-carboxylic acid (DL-ATC) to L-cysteine, subsequent oxidization of L-cysteine to L-cystine and its purification. The cells of Pseudomonas putida TS1138 could be repetitively used as the enzyme sources to convert the substrate DL-ATC to L-cysteine. After being oxidated by 2% dimethy-sulforide (DMSO), L-cystine could be harvested and further purified by the positive ion-exchange resin 001×7. High Performance Liquid Chromatography (HPLC) identified the purified L-cystine as having a total recovery of 78.55% and purity of 99.12%. This study demonstrated an efficient and convenient method for L-cystine production, which overcame the instability of enzymes, troublesome procedures and high cost of enzyme immobilization as contrasted to the traditional method. All in all, it provides a new approach for industrial production of L-cystine as well as L-cysteine. Keywords: Pseudomonas putida, DL-ATC, L-cysteine, L-cystine, Production processL-半胱氨酸是一种重要的含硫氨基酸, 广泛应用于医药、食品、化妆品以及饲料工业[1]。目前, 工业上主要通过毛发酸水解生产L-半胱氨酸。该工艺生产率低, 且产生大量刺激性气体和废酸造成环境污染[2]。另外, 由毛发水解法或其它方法获得的含半胱氨酸反应液, 由于组成复杂[], 并且半胱氨酸在水中的溶解度较高[], 所以要从复杂的反应液中分离出高纯度的半胱氨酸是困难的。半胱氨酸易被氧化成胱氨酸, 而胱氨酸在水中的溶解度极低[], 通常利用这一特性可将反应液中的半胱氨酸首先转化成胱氨酸, 沉淀析出, 然后再将分离出的胱氨酸电解还原成半胱氨酸[], 从而可获得半胱