(±)-N-(2,6-二甲苯基)-丙氨酸甲酯的脂肪酶酶促水解拆分方法  

The lipase catalyzed reaction of methyl (±)-N-(2,6-dimethylphenyl) alanine and determination of conversion

作　　者：黄丽琴[1] 杨红[1] 

机构地区：[1]南京农业大学理学院,江苏南京210095

出　　处：《南京农业大学学报》2007年第4期135-139,共5页Journal of Nanjing Agricultural University

摘　　要：酶法拆分（±）-N-（2,6-二甲苯基）-丙氨酸甲酯,其反应体系为：在含1 mmol（±）-N-（2,6-二甲苯基）-丙氨酸甲酯的100 mL 0.2 mol.L-1磷酸缓冲液（PBS）中,添加2 g聚乙二醇（PEG-6000）,并用皱落假丝酵母脂肪酶（Candida rugosalipases,CRL）拆分,反应后分离得到R-（＋）-N-（2,6-二甲苯基）-丙氨酸甲酯。进一步采用高效液相色谱检测酶促反应的转化率,采用250 mm×4.6 mm,5μHypersil（ODS色谱柱,以甲醇与水混合液（体积比为80∶20）为流动相,230 nm为检测波长,外标法峰面积定量。（±）-N-（2,6-二甲苯基）-丙氨酸和其甲酯的回收率在91.5%-96.1%之间。An enzymatic method for the production of methyl R-（ ＋ ）-N-（2,6-dimethylphenyl） alanine has been developed. The process was catalyzed by Candida rugosa lipases （ CRL）, which contained 1 mmol of methyl （ ± ） -N-（ 2,6-dimethylphenyl ） alanine, 100 mL of 0. 2 mol · L^-1 phosphate buffer solution （PBS）, as well as 2 g of polyethylene glycol （PEG-6000）, methyl R- （ ＋ ）-N-（2,6-dimethylphenyl） alanine was separated from the reaction. The concentration of（ ± ）-N-（2,6-dimethylphenyl） alanine and its methyl ester were determined with high-performance liquid chromatography on a 250 mm × 4. 6 mm, 5 μHypersil ODS column using methanol ＋ water （V： V = 80： 20） as the mobile phase, UV detection at 230 nm, and an external standard allowing quantification by measurement of peak area. The recovery was 91.5%-96. 1%. The method was also used to determine the conversion of the enzyme-catalyzed reaction.

关 键 词：(±)-N-(2 6-二甲苯基)-丙氨酸甲酯 皱落假丝酵母脂肪酶 拆分 转化率 HPLC 

分 类 号：O625.11[理学—有机化学]