生物发酵产丁二胺的定量测定  

A quantitative method for microbial-based putrescine determination

作　　者：黄荻萱 李国辉 毛银 赵运英[1,2] 周胜虎 邓禹[1,2] HUANG Dixuan;LI Guohui;MAO Yin;ZHAO Yunying;ZHOU Shenghu;DENG Yu(National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学,粮食发酵工艺与技术国家工程实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与发酵工业》2020年第12期231-236,共6页Food and Fermentation Industries

基　　金：国家重点研发计划项目(2018YFA0901400);国家自然科学基金(21877053);江苏省自然科学基金(BK20181345);江苏高校品牌专业建设工程资助项目(TAPP);江南大学自主科研-重点项目资助(JUSRP51705A,JUSRP11964)。

摘　　要：丹磺酰氯作为衍生化试剂被用于微生物发酵产丁二胺的定量测定,然而实验发现,发酵液中的复杂成分会对检测产生严重干扰,导致结果显著偏差。通过解析发酵液成分对丁二胺检测的影响规律,更加准确地实现了微生物发酵产丁二胺的测定。在丹磺酰氯柱前衍生基础上,利用校准曲线进行校准以消除误差,成功地建立了一种定量检测发酵液中丁二胺含量的方法,其检测误差<7. 96%,可消除常规检测过程中的干扰。检测方法的改进,解决了丁二胺生物合成过程中的产物难以准确定量的问题,为生产菌株的性能测定及丁二胺的生物合成奠定了基础。Dansyl chloride is used as a derivatization reagent for the quantitative determination of putrescine produced by microbial fermentation.However,the determination of putrescine was significantly interfered by the complex culture broth,leading to significant deviations in the results.The influence of fermentation broth on the detection of putrescine was studied to achieve more accurate determination of putrescine.A quantitative method using the calibration curves was successfully established with the detection error less than 7.96%,based on the precolumn derivatization of dansyl chloride.The noteworthy improvement was that it could significantly eliminate the interfering effects from culture broth without removing impurities,thus offering a feasible solution for quantitative determination of biosynthesized putrescine,and laying a solid foundation for strains selection and further industrial biosynthesis of putrescine.

关 键 词：丁二胺 发酵培养基 转化系数 校准曲线 高效液相色谱法 

分 类 号：TQ920.6[轻工技术与工程—发酵工程] TQ226.31