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中国农学通报 ›› 2012, Vol. 28 ›› Issue (12): 93-98.doi: 10.11924/j.issn.1000-6850.2011-3605

所属专题: 生物技术

• 农学 农业基础科学 • 上一篇    下一篇

溶菌酶的活性检测方法

宋美娜 仝其根   

  • 收稿日期:2011-12-02 修回日期:2012-02-09 出版日期:2012-04-25 发布日期:2012-04-25
  • 基金资助:

    北京市教委科研项目“蛋品安全生产标准体系的建立及废弃物综合利用研究”

Measuration of Lysozyme Activity

  • Received:2011-12-02 Revised:2012-02-09 Online:2012-04-25 Published:2012-04-25

摘要:

为解决现行溶壁微球菌(比浊)法测定溶菌酶活性方法中存在的测定条件不统一、重复性不高等问题。本研究对该方法中4个主要影响因素——溶菌酶活性范围、底物悬浮液吸光度值范围、检测温度和菌体制备方法进行了研究,以得到最佳的检测条件及菌体制备方法。结果表明:(1)溶菌酶活性75~250 U/mL之间,底物悬浮液吸光度值0.45~0.77 (OD450)之间,酶活力曲线具有良好的线性关系,与国家药典中基本一致;(2)检测温度25~50℃之间,底物悬浮液的吸光度变化值△OD450随温度升高而增大,适当提高检测温度可提高检测的精度,有待进一步研究;(3)用于大规模样品检测时,冷冻菌体法为制备底物悬浮液的最佳方法,检测具有良好的线性关系、管间重复性及储存稳定性。

关键词: 关键词 富硒大豆肽 胶囊 溶出特性, 关键词 富硒大豆肽 胶囊 溶出特性

Abstract:

To address the current problems of Micrococcus lysodeikticus Fleming (turbidity) method for the determination of Lysozyme activity,such as not unified measurement conditions, poor repeatability, four main influence factors have been studied which were respectively the range of Lysozyme activity, substrate suspension absorbance value range, measuring temperature and cell preparation in this paper. Ultimately, the study expected to be the best test conditions and substrate preparation method. The results were as follows: (1) the Lysozyme activity curves had good linear relation when the Lysozyme active range of 75 to 250 U/mL and substrate suspension absorptance value range of 0.45-0.77 (wavelength 450 nm),in general accord with the GBT 25879-2010; (2) In measuring temperature 25~50℃, substrate suspensions of absorbance difference (△OD450) increased with the rise of temperature. Appropriate to improve the measuring temperature can improve the measuring accuracy, which is needed to be further studied; (3) For large-scale sample test, frozen bacteria was selected as the optimum substrate, which had good linear relation, better repeatability and storage stability.

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