Gong P-F, Wu H-Y, Xu J-H, Shen D, Liu Y-Y
Laboratory of Applied Biocatalysis, P. O. Box 283, State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, 130 Meilong Road, Shanghai 200237, P. R. China.
Appl Microbiol Biotechnol. 2002 May;58(6):728-34. doi: 10.1007/s00253-001-0860-0. Epub 2002 Mar 15.
The yeast strain CGMCC 0573 was identified as Citeromyces matriensis and shown to be capable of enantioselectively hydrolyzing ethyl ester of ( R)-Ketoprofen (2-(3-benzoylphenyl)propionic acid). The strain was isolated for the first time from soil samples through a new and efficient screening procedure in which the probability of obtaining active strains was greatly increased by using ethanol and Tween-80 alternatively as additives during the enrichment culture. Studies of the culture conditions and catalytic performance of Citeromyces matriensis CGMCC 0573 showed that the enzyme occurs constitutively in the cells and its production is enhanced by feeding with Tween-80 during the early period of cultivation. Yeast extract was found to be beneficial both for growth and for esterase production. The optimal temperature and pH for the bioconversion were 40 degrees C and pH 8.0, respectively. Biotransformation using resting cells cultured in a flask with baffles and magnetic stirring and in the presence of 50 mM substrate resulted in the production of ( R)-ketoprofen at 93% ee (enantiomeric excess) and at 42.6% conversion.
酵母菌株CGMCC 0573被鉴定为母生 Citromyces matriensis,并被证明能够对映选择性水解(R)-酮洛芬(2-(3-苯甲酰基苯基)丙酸)的乙酯。该菌株首次通过一种新的高效筛选程序从土壤样品中分离得到,在富集培养过程中交替使用乙醇和吐温80作为添加剂,大大提高了获得活性菌株的概率。对母生 Citromyces matriensis CGMCC 0573的培养条件和催化性能的研究表明,该酶在细胞中组成性存在,在培养初期添加吐温80可提高其产量。发现酵母提取物对生长和酯酶生产均有益。生物转化的最佳温度和pH分别为40℃和pH 8.0。使用在带挡板的烧瓶中培养并进行磁力搅拌且存在50 mM底物的静止细胞进行生物转化,得到的(R)-酮洛芬对映体过量(ee)为93%,转化率为42.6%。