用木瓜蛋白酶及固定化木瓜蛋白酶拆分DL_苯丙氨酸_英文_

ISSN 1007 7626CN 11 3870 Q

中国生物化学与分子生物学报

Chinese Journal of Biochemistry and Molecular Biology

2009年1月25(1):23~29

Resolution of DL Phenylalanine by Papain and Immobilized Papain

HE Ping,HUANG Zhuo Lie *

,W U Guang Hong,C HU Zhi Zhan,SHI Qing Cui,HUANG Jia Le

(Colle ge o f Li fe Sciences ,South China Agricultural U ni versity ,Guangzhou 510642,China )

Abstract In order to separate chiral D phenylalanine from L phenylalanine,the enzyme catalytic method by papain and immobilized papain were used in this investigation.It indicated that the yield rate of N acetyl DL phenylalanine synthesized from DL phenylalanine was 88 7%.The ionic strength was the strongest influencing factor in the synthesis of N acetyl L phenylalanylaniline by both papain and immobilized papain on sodium alginate chitosan (IPSAC).But the reaction temperature was the strongest influencing factor in the synthesis by immobilized papain on nylon cloth (IPN).The ionic strength and the pH were the important influencing factors in the synthesis.The yield rates of the best synthesis system of N acetyl L phenylalanylaniline by papain,IPSAC,IPN were 61 2%,54 7%,36 3%,respectively.The yield rate of L phenylalanine from N acetyl L phenylalanylaniline hydrolyzation was 59 2%.The optical purity of the product was 96 6%.The yield rate of D phenylalanine from N acetyl D phenylalanine hydrolyzation was 61 7%.The optical purity was 95 7%.

Key words immobilized papain;DL phenylalanine;resolution;optical purity

用木瓜蛋白酶及固定化木瓜蛋白酶拆分DL 苯丙氨酸

何 平, 黄卓烈*

, 巫光宏, 初志战, 史清翠, 黄家乐

(华南农业大学生命科学学院,广州 510642)

摘要 为了将手性化合物D 苯丙氨酸和L 苯丙氨酸进行分离,利用木瓜蛋白酶及固定化木瓜蛋白酶催化的方法对其拆分.试验结果表明,用DL 苯丙氨酸合成N 乙酰 DL 苯丙氨酸,得率为88 7%.木瓜蛋白酶、海藻酸钠 壳聚糖固定化木瓜蛋白酶(IPSAC)、尼龙布固定化木瓜蛋白酶(IPN)催化合成N 乙酰 L 苯丙氨酰苯胺时,对催化合成过程影响最大的因素分别是溶液中的离子强度、溶液中的离子强度、反应温度;溶液中的离子强度与pH 对合成的影响较大.本试验得出分别用木瓜蛋白酶、IPSAC 、IPN 催化合成N 乙酰 L 苯丙氨酰苯胺的3个最佳方案;用此3个方案合成时,产率分别为61 2%、54 7%、36 3%.N 乙酰 L 苯丙氨酰苯胺水解生成L 苯丙氨酸,产率59 2%,光学纯度为96 6%.N 乙酰 D 苯丙氨酸水解生成D 苯丙氨酸,产率61 7%,光学纯度为95 7%.关键词 固定化木瓜蛋白酶;DL 苯丙氨酸;拆分;光学纯度中图分类号 Q556

Recei ved:June 17,2008;Accepted:July 29,2008

Supported by National Natural Science Foundation of China (No.10074016)and Guangdong Natural Science Foundation(No.7006655)

*

Corres pondi ng author Tel:020 ********;

E mail:zhuolieh@s

收稿日期:2008 06 17;接受日期:2008 07 29

国家自然科学基金(No.10074016)和广东省自然科学基金(No.7006655)资助

*

联系人 Tel:020 ********;E mail:zhuolieh@s

Chirality of compound is natural phenomenon.The main amino acids are L type in life.About 40%

medicines are chiral compounds [1]

.L phenylalanine is one of the indispensable a mino acids for people and animals.Also it is the materials of synthesizing the medication of resisting virus and cancer,and materials of synthesizing new type sweets.Whereas D phenylalanine

can enforce human immunity and relieve pain finely [2]

.At present,besides dissymmetric synthesis and ferment,chiral compounds are obtained by the resolution of the racemic mixture of chemosynthesis.The main resolution methods of chiral compounds were crystal resolution,chromatography resolution,enzyme resolution,chemistry

resolution,e xtrac tion resolution and others [3~8]

.

Recently,chiral medicines are produced

by