新型假单胞菌对羟基苯乙腈水解酶及其性质研究.docVIP

Bioprocess 生物过程, 2012, 2, 70-74 /10.12677/bp.2012.22012 Published Online June 2012 (/journal/bp) A New 4-Hydroxyphenylcyanide Nitrilases from Pseudomonas sp. and Its Catalytic Properties* Mingle Cao , Xinglin Jiang , Haibo Zhang , Mo Xian , Xin Xu , Wei Liu 2 1 1# 1 1 1 1 Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao 2 State Key Laboratory of Microbial Technology, Shandong University, Ji’nan # Email: zhanghb@ Received: May 13th, 2012; revised: May 27th, 2012; accepted: Jun. 4th, 2012 Abstract: Nitrilases can hydrolyze nitrile efficiently under mild conditions. The enzymatic methods have the advan- tages of less pollution, low cost compared with the chemical methods. Nitrilases are potentially applied in agriculture, industry, environment, and biomedicine. In this study, Berthelot method and high performance liquid chromatography were used to screen new stains for nitrilases. A nitrilase with high substrate specificity for 4-hydroxyphenylcyanide was found from Pseudomonas sp. 6-1. The stain produced 28.47 U/mL nitrilase after optimization of the culture conditions. The enzyme remained 80% activities at pH 6.6 to pH 7.6, temperature 35?C to 45?C, and it was stable after 18 h incuba- tion. The stain can be potentially used in biosynthesis 4-Hydroxyphenylacetic acid using 4-hydroxyphenylcyanide as substrate. Keywords: 4-Hydroxyphenylcyanide; 4-Hydroxyphenylacetic Acid; Nitrilase; Biocatalysis 新型假单胞菌对羟基苯乙腈水解酶及其性质研究* 曹明乐，姜兴林，张海波，咸漠，徐鑫，刘炜 1 2 1 1# 1 1 1 中国科学院青岛生物能源与过程研究所，青岛 2 山东大学微生物技术国家重点实验室，济南 Email: zhanghb@ # 收稿日期：2012年 5月 13日；修回日期：2012年 5月 27日；录用日期：2012年 6月 4日 摘 要：腈水解酶催化的腈水解具有反应高效、条件温和、环境污染小和成本低等优点，在有机合成、材料合 成、医药、食品、农业、畜牧业及环境等污染方面有着重要的应用前景。本研究利用初步活化和分步胁迫富集， 通过 Berthelot法高通量筛选与高压液相精细筛选获得一株底物对 4-羟基