重组色氨酸合成酶催化合成L-5-羟基色氨酸
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摘要
利用重组色氨酸合成酶催化合成L-5-羟基色氨酸,采用单因素以及响应面分析方法考察了p H、反应温度、底物浓度和底物摩尔比对L-5-羟基色氨酸合成的影响。最佳转化条件为:反应温度为32℃,p H=8.6,5-羟基吲哚与工业角蛋白水解氨基酸液中L-丝氨酸的适宜底物摩尔比为1.1∶1,底物工业角蛋白水解氨基酸液中L-丝氨酸最适合浓度为180 mmol/L,反应平衡时间为18 h,工业角蛋白水解氨基酸液中L-丝氨酸摩尔转化率达到86.5%。
Enzymatic synthesis of L-5-hydroxytryptophan from L-serine and 5-hydroxyindole catalyzed by tryptophan synthase from recombinant Escherichia coli were studied.The factors such as temperature,p H,molar ratio of L-serine to 5-hydroxyindole and substrate concentration were investigated by single-factor experiments and response surface methodology.The optimal temperature and p H value were 32 ℃ and 8.6,respectively.The optimal molar ratio of 5-hydroxyindole to L-serine was 1.1 ∶ 1.The optimal substrate concentration of L-serine was 180 mmol / L.Under the optimal conditions,the bioconversion rate of L-serine reached 86.5% after 18 h.
Enzymatic synthesis of L-5-hydroxytryptophan from L-serine and 5-hydroxyindole catalyzed by tryptophan synthase from recombinant Escherichia coli were studied.The factors such as temperature,p H,molar ratio of L-serine to 5-hydroxyindole and substrate concentration were investigated by single-factor experiments and response surface methodology.The optimal temperature and p H value were 32 ℃ and 8.6,respectively.The optimal molar ratio of 5-hydroxyindole to L-serine was 1.1 ∶ 1.The optimal substrate concentration of L-serine was 180 mmol / L.Under the optimal conditions,the bioconversion rate of L-serine reached 86.5% after 18 h.
引文
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[2]Kema I P,De Vries E G E,Muskiet F A J.Clinical chemistry of serotonin metabolites[J].J Chrom B,2000,747:33-48.
[3]Ohgi Y,Futamura T,Kikuchi T,et al.Effects of antidepressants on alternations in serum cytokines and depressive-like behavior in mice after lipopolysaccharide administration[J].Pharmacol Biochem Behav,2013,103(4):853-859.
[4]Gendle M H,Young E L,Romano A C.Effects of oral 5-hydroxytryptophan on a standardized planning task:insight into possible dopamine/serotonin interactions in the forebrain[J].Hum Psychopharmacol,2013,28(3):270-273.
[5]Jangid P,Malik P,Singh P,et al.Comparative study of efficacy of1-5-hydroxytryptophan and fluoxetine in patients presenting with first depressive episode[J].Asian J Psychiatr,2013,6(1):29-34.
[6]徐家业.有机合成化学及近代技术[M].西安:西北工业大学出版社,1997:108.
[7]马冬梅.5-羟基色氨酸的制备及分析方法研究[D].西安:西北大学,2006.
[8]李玉山.从非洲加纳籽提取、纯化5-羟基色氨酸的工艺[J].计算机与应用化学,2010,27(7):932-936.
[9]聂利芳.从加纳籽中提取、分离纯化5-羟基色氨酸的工艺研究[D].西安:西北大学,2009.
[10]冯建光.离子交换树脂分离纯化加纳籽中5-羟基色氨酸[J].中草药,2013,44(17):2410-2416.
[11]Ye L,Hu Z P,Du G Y,et al.Antidepressant-like effects of the extract from Cimicifuga foetida L[J].J Ethnopharmacol,2012,144(3):683-691.
[12]王建军.利用工程菌株BL21-DE3发酵获取5-羟基色氨酸的方法[P].CN:201010183157.8,2008.
[13]Bornscheuer U T,Pohl M.Improved biocatalysts by directed evolution and rational protein design[J].Current Opinion in Chemical Biology,2001,5:137-143.
[14]萨姆布鲁克J,拉塞尔D W.分子克隆实验指南[M].3版.北京:科学出版社,2002.
[15]Xu Lisheng,Wang Zhiyuan,Liu Junzhong,et al.Effects of polyhydroxy compounds on enzymatic synthesis of L-tryptophan catalyzed by tryptophan synthase[J].Catalysis Letters,2012,142(2):282-286.
[16]欧艳,肖方青,肖艳萍.5-羟基色氨酸的提取和测定方法研究[J].湖南中医杂志,2005(4):83-84.