L-半胱氨酸产生菌恶臭假单胞菌TS1138的鉴定和诱变育种  

Identification and Mutation Breeding of Pseudomonas Putida TS1138 with Capability of Biosynthesizing L-Cysteine from ATC

作　　者：孙丹[1] 余养盛[2] 杨文博[2] 田旺[2] 白钢[2] 

机构地区：[1]南开大学学报编辑部,天津300071 [2]南开大学生命科学学院,天津300071

出　　处：《天津大学学报》2007年第4期421-426,共6页Journal of Tianjin University(Science and Technology)

基　　金：国家自然科学基金资助项目(30470053);天津市重点基金资助项目(05YFJZJC00900)

摘　　要：从土壤中分离得到一株利用DL-2-氨基-Δ2-噻唑啉-4-羧酸合成L-半胱氨酸的TS1138菌株,通过形态学特征、生理生化特征、(G+C)(摩尔分数)和16S rRNA基因序列对其进行了分类鉴定,确定TS1138菌株为恶臭假单胞菌(Pseudomonas putida).该菌株具有将DL-2-氨基-Δ2-噻唑啉-4-羧酸不对称地水解生成L-半胱氨酸的性能.采用了L-半胱氨酸产物的检测方法即18-磷钨酸法、薄层层析法和旋光度法.将TS1138菌株进行亚硝基胍诱变,得到TS1138菌株脱巯基酶缺失突变株TS1138-10,其产L-半胱氨酸能力较出发菌株提高了5000/以上.TS1138-10的稳定性实验表明,该菌株的高产L-半胱氨酸能力具有稳定性的遗传特性.A strain TS1138 that can synthesize L-cysteine from DL-2-amino-△^2-thiazoline-4-carboxylic acid （ATC） was isolated and enriched from the soil. Based on analyses of the phenotype, physiologic and biochemical specificity, （ G ＋ C） （ molar percentage）, and 16S rRNA gene sequence, it is confirmed that the TS1138 strain belongs to Pseudomonas putida. This strain can asymmetrically hydrolyze ATC to generate L-cysteine. The qualitative and quantitative detecting methods of L-cysteine, including method of 18-phosphotungstic acid, thin layer chromatography and method of speficic rotation, were used. A mutant strain TS1138-10 defective in cysteine desulfhydrase was obtained by N-methyl-N＇-nitro-N-nitrosoguanidine （NTG） mutation. The L-cysteine biosynthesis capability of TS1138-10 was improved more than 50%, and this enhanced L-cysteine biosynthesis property is genetically stable.

关 键 词：恶臭假单胞菌TS1138 DL-2-氨基-△^2-噻唑啉4-羧酸 L-半胱氨酸 

分 类 号：Q93-331[生物学—微生物学]