恶臭假单胞菌CGMCC1388对映选择性生物降解制备(S)-扁桃酸及其衍生物的研究  

Preparation of (S)-mandelic acid and its derivatives by enantioselective degradation of racemic mandelates with Pseudomonas putida CGMCC1388

作　　者：黄汉荣[1] 潘江[1] 徐毅[1] 许建和[1] 

机构地区：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

出　　处：《生物加工过程》2005年第3期47-51,共5页Chinese Journal of Bioprocess Engineering

基　　金：国家自然科学基金重大项目(203900506);国家973计划(2003CB716008)

摘　　要：从土壤中分离得到一株(R)-扁桃酸选择性降解菌,经鉴定为恶臭假单胞菌,保存于中国普通微生物保藏中心,编号为CGMCC1388。考察了扁桃酸及其降解产物对扁桃酸脱氢酶活力的影响。研究表明,在培养基中添加少量扁桃酸、苯甲酰甲酸或苯甲酸均可显著提高其产量,以扁桃酸的诱导效果最佳。3种诱导物的最适添加质量浓度分别为4、4和2 g/L。当以消旋扁桃酸为反应底物时,该菌可高选择性降解(R)-扁桃酸,回收得到的(S)-扁桃酸对映体过量值(e.e.)高于99%,反应的对映选择率(E)达130。用静息细胞催化扁桃酸降解的最适温度和pH分别为30℃和6.0,最适底物浓度为60 mmol/L,以双倒数法求得Km为47 mmol/L。考察了该菌对扁桃酸苯环取代衍生物的生物转化,并以高产率制备获得高对映纯度的(S)-对羟基扁桃酸和(S)-对氯扁桃酸。A mandelate-degrading bacterium, Pseudomonas putida CGMCC1388, was newly isolated from soil using benzoylformic acid as sole carbon source. It could degrade （ R）-mandelic acid with high enantioselectivity （ E = 130）. Supplement of mandelic acid, benzoylformic acid or benzoic acid to the culture medium as the enz- yme inducer significantly enhanced the degradation activity of the bacterial cells. The optimal concentrates of these inducers were 4 g/L, 4 g/L and 2 g/L respectively. The optimal temperature and pH for the mandelate degradation were 30 ℃and 6.0 respectively, and the optimal substrate concentration was 60 mmol/L.The apparent kinetic constants Km determined was 47 mmol/L. Using the resting cells as bioeatalyst, （S）-mandelic acid, （S）-p-hydroxyrnandelic acid and （S）-p-chloromandelic acid were readily prepared, with moderate to high yields and excellent enantiopurity （ 〉 99 % e. e. ）.

关 键 词：恶臭假单胞菌 扁桃酸脱氢酶 对映选择性降解 (S)-扁桃酸 (S)-扁桃酸衍生物 

分 类 号：Q814[生物学—生物工程]