基于比浊法的杆菌肽定量测定方法优化

doi:10.13560/j.cnki.biotech.bull.1985.2019-0994

生物技术通报 ›› 2020, Vol. 36 ›› Issue (5): 92-97.doi: 10.13560/j.cnki.biotech.bull.1985.2019-0994

• 生物分析方法与标准物质专题（专题主编：李亮副研究员） • 上一篇下一篇

基于比浊法的杆菌肽定量测定方法优化

王志新¹, 刘洋¹, 周景波¹, 宏丹¹, 鲁雷震¹, 宁亚维¹, 贾英民²

1.河北科技大学生物科学与工程学院河北省发酵工程技术研究中心,石家庄 050018;
2.北京工商大学食品与健康学院,北京 100048

收稿日期:2019-10-17 出版日期:2020-05-26 发布日期:2020-06-03
作者简介:王志新,女,博士,副教授,研究方向：发酵工程;E-mail：zhxwang0311@163.com
基金资助:
“十三五”河北省重点研发计划项目（18227127D）,“十三五”国家重点研发计划项目（2016YFD0400800,2016YFF0202302）

Optimization of Quantitative Determination of Bacitracin Based on Turbidimetric Method

WANG Zhi-xin¹, LIU Yang¹, ZHOU Jing-bo¹, HONG Dan¹, LU Lei-zhen¹, NING Ya-wei¹, JIA Ying-min²

1. College of Bioscience and Bioengineering,Hebei University of Science and Technology,Research Center for Fermentation Engineering of Hebei,Shijiazhuang 050018;
2. School of Food and Health Engineering,Beijing Technology and Business University,Beijing 100048

Received:2019-10-17 Published:2020-05-26 Online:2020-06-03

摘要/Abstract

摘要： 杆菌肽在研究应用过程中,定量测定方法不统一,结果缺乏参考性。为规范其测定方法,拟通过建立杆菌肽浓度对数值与OD₆₀₀之间的线性关系,以重复性和精密度为指标,优化指示菌初始浓度、杆菌肽溶液与菌悬液的比例、培养时间等因素,确定比浊法测定杆菌肽抑菌活性的方法。结果显示,比浊法的最适测定条件为：指示菌初始浓度10⁷ CFU/mL,杆菌肽溶液与菌悬液比例1∶9,培养时间4 h,在此条件下,线性关系良好,R²达到0.99以上,且具有良好的重复性。进一步选用大肠杆菌和金黄色葡萄球菌验证方法的可行性,方法重复性好,精密度高。研究结果将为比浊法的进一步应用以及杆菌肽在试验和生产过程中的定量测定提供参考和依据。

关键词: 比浊法, 定量测定, 杆菌肽, 抑菌性

Abstract: The quantitative determination methods of bacitracin in the research and application are not standardized,and the results cannot be used for reference. In order to standardize the determination methods,the linear relationship between the logarithmic value of bacitracin concentration and OD₆₀₀ was established,and the initial concentration of indicating bacteria,the ratio of bacitracin solution to bacterial suspension and the culture time were optimized with repeatability and precision as indicators. The optimal determination conditions of bacitracin against bacteria by turbidimetric method were as follows：the initial concentration of indicating bacteria were 10⁷ CFU/mL,the ratio of bacitracin solution to bacterial suspension was 1∶9,and the culture time was 4 h. Under these conditions,the linear relationship was fine with R² above 0.99 and the repeatability was well. Furthermore,Escherichia coli and Staphylococcus aureus were selected to verify the feasibility of the method,and this method presented favorable repeatability and high precision. The results would provide reference and basis for the further application of turbidimetric method and the quantitative determination of bacitracin in the test and production process.

Key words: turbidimetric method, quantitative determination, bacitracin, antibacterial activity

王志新, 刘洋, 周景波, 宏丹, 鲁雷震, 宁亚维, 贾英民. 基于比浊法的杆菌肽定量测定方法优化[J]. 生物技术通报, 2020, 36(5): 92-97.

WANG Zhi-xin, LIU Yang, ZHOU Jing-bo, HONG Dan, LU Lei-zhen, NING Ya-wei, JIA Ying-min. Optimization of Quantitative Determination of Bacitracin Based on Turbidimetric Method[J]. Biotechnology Bulletin, 2020, 36(5): 92-97.

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基于比浊法的杆菌肽定量测定方法优化

Optimization of Quantitative Determination of Bacitracin Based on Turbidimetric Method

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