利用纸层析-分光光度法检测甲基营养菌5222产L-丝氨酸  

Detection of L-serine produced by Methylobacterium sp.5222 by paper chromatography-spectrophotometry

作　　者：张建云 谷立坤 崔芽芽 韩梦霞 彭更 史永胜 ZHANG Jianyun;GU Likun;CUI Yaya;HAN Mengxia;PENG Geng;SHI Yongsheng(College of Resource and Environment,Henan University of Engineering,Zhengzhou 451191,China;Henan Engineering Laboratory of Pollution Control and Coal Chemical Resources Comprehensive Utilization,Henan University of Engineering,Zhengzhou 451191,China)

机构地区：[1]河南工程学院资源与环境学院,河南郑州451191 [2]河南工程学院煤化工资源综合利用与污染治理河南省工程实验室,河南郑州451191

出　　处：《中国酿造》2019年第11期114-118,共5页China Brewing

基　　金：河南工程学院博士基金资助项目(D2010014);煤化工资源综合利用与污染治理河南省工程实验室开放基金资助项目(502002-B04)

摘　　要：该研究旨在建立一种准确、简便的检测L-丝氨酸含量的方法。当纸层析-分光光度法展开剂为丙酮∶正丁醇∶二乙胺∶水(10∶10∶2∶5,V/V),pH值为12.7,波长为510 nm,洗脱时间为30 min时,能很好的对L-丝氨酸进行定性、定量检测。当L-丝氨酸标准溶液质量浓度在1~6μg/mL时,L-丝氨酸的含量(x)与吸光度值(y)之间有良好的线性相关关系(回归方程y=0.0088x+0.0089,R2=0.9962);精密度实验结果显示相对标准偏差(RSD)为2.3%;回收率实验结果显示平均加标回收率95.3%,RSD为2.6%。说明该方法具有较高的精密度和准确度。以甲基营养型菌(Methylobacterium sp.)5222为出发菌株,以甘氨酸为前体物质发酵生产L-丝氨酸,应用该研究建立的纸层析-分光光度法,检测到发酵液中L-丝氨酸含量为0.90 g/L,说明菌株5222具有产L-丝氨酸的能力。The aim of the study was to establish an accurate and simple method for the detection of L-serine content.When the developing solvent for chromatography-spectrophotometry was acetone∶n-butanol∶diethylamine∶water(10∶10∶2∶5,V/V),the pH was 12.7,the wavelength was 510 nm,and the elution time was 30 min,the L-serine could be qualitatively and quantitatively detected.When the concentration of the L-serine was 1-6μg/ml,there was a good linear relationship(regression equation y=0.0088+0.0089x,R2=0.9962)between L-serine content(x)and absorbance value(y).The precision test results showed that the relative standard deviation(RSD)was 2.3%.The recovery rate test results showed that the average standard recovery rate was 95.3%,and the RSD was 2.6%,showing that the method had high precision and accuracy.Using the Methylobacterium strain 5222 as original strain,and glycine as a precursor,L-serine was produced,and the L-serine content in the fermentation broth was 0.90 g/L detected by paper chromatography-spectrophotometry,indicating that the strain 5222 had the ability to produce L-serine.

关 键 词：纸层析-分光光度法 L-丝氨酸 甲基营养菌 检测 

分 类 号：TQ922[轻工技术与工程—发酵工程]