冬虫夏草中核苷酸酶和腺苷脱氨酶的酶学特征及来源溯源

IF 4.6 2区 农林科学 Q2 CHEMISTRY, APPLIED
Wenqing Li, Baoqing Wen, Kunxia Lin, Wenjia Li, Zhengming Qian
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引用次数: 0

摘要

腺苷是珍贵真菌冬虫夏草(c.s inensis)的重要品质标志。本文首次建立了以高效液相色谱(hplc)为基础,定量测定中华绒猴核酸酶和腺苷脱氨酶两种腺苷代谢酶活性和动力学参数的方法。优化了酶提取方案、反应体系和分析程序。选择水作为提取溶剂,以保持天然酶活性。核苷酸酶在pH 6和35°C时表现出最佳活性,而腺苷脱氨酶在pH 8和45°C时表现出最佳活性,显示出不同的催化谱。与核苷酸酶相比,腺苷脱氨酶表现出更好的热稳定性。动力学分析表明,核苷酸酶的Michaelis常数为0.24µmol·mL−1,最大反应速率为7.92µmol·L−1·min−1,而腺苷脱氨酶的Michaelis常数为0.14µmol·mL⁻¹ 和0.61µmol·L⁻¹ ·min⁻¹ 。方法验证证实了分析系统的可靠性。来源溯源分析表明,核苷酸酶主要来源于真菌成分,而腺苷脱氨酶仅来源于寄主幼虫,仅在昆虫组织中检测到。因此,腺苷脱氨酶活性可作为区分无虫真菌和菌虫复合体的可靠标志。这些研究结果为优化中华香椿的腺苷积累、提高其质量控制和管理水平提供了科学依据。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Enzymatic characteristics and origin tracing of nucleotidase and adenosine deaminase in Cordyceps sinensis based on HPLC
Adenosine is a key quality marker for the valuable fungus Cordyceps sinensis (C. sinensis). This study presents the first HPLC-based method for quantifying the activities and kinetic parameters of two adenosine-metabolizing enzymes—nucleotidase and adenosine deaminase—in C. sinensis. The enzyme extraction protocols, reaction systems, and assay procedures were optimized. Water was selected as the extraction solvent to preserve native enzymatic activity. Nucleotidase exhibited optimal activity at pH 6 and 35°C, while adenosine deaminase showed optimal activity at pH 8 and 45°C, revealing distinct catalytic profiles. Adenosine deaminase exhibited superior thermostability compared to nucleotidase. Kinetic analysis revealed that nucleotidase had a Michaelis constant of 0.24 µmol·mL−1 and a maximum reaction rate of 7.92 µmol·L−1·min−1, while adenosine deaminase showed corresponding values of 0.14 µmol·mL⁻¹ and 0.61 µmol·L⁻¹ ·min⁻¹ . Method validation confirmed the reliability of the analytical system. Source-tracing analysis revealed that nucleotidase originated primarily from the fungal component, whereas adenosine deaminase was exclusively derived from the host larva and detected only in insect tissues. Therefore, adenosine deaminase activity serves as a reliable marker for distinguishing insect-free fungus from fungus-insect complex. These findings provide a scientific basis for optimizing adenosine accumulation and improving the quality control and management of C. sinensis.
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来源期刊
Journal of Food Composition and Analysis
Journal of Food Composition and Analysis 工程技术-食品科技
CiteScore
6.20
自引率
11.60%
发文量
601
审稿时长
53 days
期刊介绍: The Journal of Food Composition and Analysis publishes manuscripts on scientific aspects of data on the chemical composition of human foods, with particular emphasis on actual data on composition of foods; analytical methods; studies on the manipulation, storage, distribution and use of food composition data; and studies on the statistics, use and distribution of such data and data systems. The Journal''s basis is nutrient composition, with increasing emphasis on bioactive non-nutrient and anti-nutrient components. Papers must provide sufficient description of the food samples, analytical methods, quality control procedures and statistical treatments of the data to permit the end users of the food composition data to evaluate the appropriateness of such data in their projects. The Journal does not publish papers on: microbiological compounds; sensory quality; aromatics/volatiles in food and wine; essential oils; organoleptic characteristics of food; physical properties; or clinical papers and pharmacology-related papers.
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