微生物学通报 JAN 20，2008，35(1)：45~49
(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 300222)
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, 并进一步氧化和分离纯化产物L-胱氨酸的生产工艺和条件进行了研究。建立了以恶臭假单胞菌TS1138 (Pseudomonas putida TS1138)全细胞为酶源, 反复多次催化底物合成L-半胱氨酸, 并以2.0%二甲基亚砜(DMSO)为氧化剂氧化生成L-胱氨酸, 进而通过001×7型阳离子交换树脂纯化胱氨酸的新工艺。采用高效液相色谱法考察该方法L-胱氨酸的总收率可以达到78.55%, 纯度为99.12%。该方法简单高效, 解决了酶稳定性差不能重复使用, 而固定化酶方法繁琐成本高的问题, 为我国L-半胱氨酸和L-胱氨酸的生产开辟一条新途径。
关键词: 恶臭假单胞菌, DL-ATC, L-半胱氨酸, L-胱氨酸, 生产工艺
Study on L-cystine Conversion Technology by Pseudomonas putida TS1138
LIU Chun-Qin1 YU Yang-Sheng1 BAI Gang1* YANG Wen-Bo1
CHEN 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 enzy-matic conversion of DL-2-amino-∆2-thiazoline-4-carboxylic acid (DL-ATC) to L-cysteine, subsequent oxi-dization 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 pro-vides 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
全文链接：恶臭假单胞菌TS1138转化生产L-胱氨酸的工艺研究
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