高效液相色谱法检测肺炎克雷伯氏菌发酵液中的3-羟基丙酸  

Detection of 3-hydroxypropionic in fermentation broth of Klebsiella pneumoniae by high performance liquid chromatography

作　　者：田甜 张炎 平文祥[1] 葛菁萍[1,2] TIAN Tian;ZHANG Yan;PING Wen-Xiang;GE Jing-Ping(Heilongjiang University Key Laboratory of Microbiology,Life Science College,Harbin 150080,China;Heilongjiang UniversityEngineering Research Center of Agricultural Microbiology Technology,Ministry of Education,Harbin 150500,China)

机构地区：[1]黑龙江大学生命科学学院微生物省高校重点实验室,哈尔滨150080 [2]黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨150500

出　　处：《黑龙江大学工程学报》2022年第4期84-89,共6页Journal of Engineering of Heilongjiang University

基　　金：黑龙江大学研究生创新科研项目(YJSCX2021-202HLJU);国家自然科学基金面上项目(32071519);黑龙江省自然科学基金重点项目(ZD2020C008)。

摘　　要：3-羟基丙酸(3-hydroxypropionic acid,3-HP)是一种重要的平台化合物。肺炎克雷伯氏菌(Klebsiella pneumoniae)利用甘油生产3-HP的过程中,常伴有乳酸等副产物的产生。在利用高效液相色谱法(high performance liquid chromatography,HPLC)检测3-HP产量时,两者色谱峰重叠、难以有效检测3-HP产量。因此,通过选择与优化HPLC的检测条件,建立了一种简单、快速、准确检测3-HP含量的检测方法。结果表明:利用色谱柱HPX-87H(300 mm×7.8 mm,5μm),流动相5 mmol·L^(-1)H_(2)SO_(4),流速0.6 mL·min^(-1),柱温65℃,进样量20μL,采用紫外检测器(210 nm),完成了3-HP产量的检测。该方法不仅提高了3-HP的检测效率和检测精度,还可以准确检测发酵过程中产物的变化。3-Hydroxypropionic acid(3-HP)is an important platform chemical.The production of 3-HP from glycerol by Klebsiella pneumoniae using glycerol is often accompanied by the production of by-products such as lactic acid.When using HPLC to detect 3-HP yield,the two chromatographic peaks overlap,so it was difficult to detect 3-HP yield effectively.Therefore,by selecting and optimizing the detection conditions of HPLC,a simple,rapid and accurate method for the detection of 3-HP content was established.The results showed that the detection of 3-HP yield was accomplished by using HPX-87H column(300 mm×7.8 mm,5μm),and the mobile phase was 5 mmol·L^(-1)of sulfuric acid solution,flow rate was 0.6 mL·min^(-1),column temperature was 65℃,injection volume was 20μL,ultraviolet detector(210 nm)was used.This method not only improves the efficiency and accuracy of 3-HP detection,but also allows accurate detection of changes in products during fermentation.

关 键 词：3-羟基丙酸 肺炎克雷伯氏菌 高效液相色谱法 发酵液 

分 类 号：Q939.99[生物学—微生物学]