六神曲发酵过程中酶学性质与微生物群落变化相关性研究  

Correlation between changes in enzymatic properties and microbial communities during fermentation of Massa Medicata Fermentata

作　　者：丁海玲 时海燕[2] 王爽 徐丹丹 赵霞 崔彦伟 徐男 DING Hailing;SHI Haiyan;WANG Shuang;XU Dandan;ZHAO Xia;CUI Yanwei;XU Nan(School of Medicine,Qilu Institute of Technology,Shandong Jinan 250299,China;Department of Clinical Pharmacy,First Affiliated Hospital,Shandong First Medical University&Shandong Provincial Qianfoshan Hospital,Shandong Engineering&Technology Research Center for Pediatric Drug Development,Shandong Medicine and Health Key Laboratory of Clinical Pharmacy,Shandong Jinan 250014,China;School of Pharmacy,Shandong University of Traditional Chinese Medicine,Shandong Jinan 250355,China;Laboratory of Chinese Medicine Preparation,Shandong Research Academy of Traditional Chinese Medicine,Shandong Jinan 250014,China;R&D Division,Shandong Baiweitang Traditional Chinese Medicine Slices Inc,Shandong Jinan 250100,China)

机构地区：[1]齐鲁理工学院医学院,山东济南250299 [2]山东第一医科大学第一附属医院(山东省千佛山医院)临床药学部,山东省儿童药物临床评价与研发工程技术研究中心,山东省医药卫生临床药学重点实验室,山东济南250014 [3]山东中医药大学药学院,山东济南250355 [4]山东省中医药研究院中药制剂研究所,山东济南250014 [5]山东百味堂中药饮片有限公司研发部,山东济南250100

出　　处：《中国医院药学杂志》2024年第17期1980-1986,共7页Chinese Journal of Hospital Pharmacy

基　　金：山东省科技型中小企业创新能力提升工程(编号:2022TSGC1057);济南市科技型中小企业创新能力提升工程(编号:2022TSGC1057);济南市“新高校20条”引进创新团队项目(编号:202333021);山东省重点研发计划重大科技创新工程(编号:2022CXGC020514);山东省自然科学基金重点项目(编号:ZR2020KH017)。

摘　　要：目的:以六神曲为研究对象,对发酵过程中酶学性质与微生物群落变化之间的相关性进行研究。方法:采用酶活检测方法对六神曲酶学特性进行评价,采用MiSeq高通量测序技术对微生物类群进行解析,利用相关性分析发酵过程中酶的性质变化与微生物群落之间的相关性。结果:发酵初期,微生物大量繁殖,淀粉酶、糖化酶、蛋白酶、脂肪酶、滤纸酶和纤维素酶等活性指标急剧上升;发酵中末期,微生物种类下降,曲霉菌占据主导地位。糖化酶活性在36 h达到最高值53.818μmol·g^(-1),淀粉酶在30 h达到最高值87.841μmol·g^(-1),脂肪酶、滤纸酶和纤维素酶在24 h分别达到最高值0.048μmol·g^(-1)、1.623μmol·g^(-1)和1.521μmol·g^(-1),均呈现先升后降的变化趋势;经过42 h,蛋白酶开始呈现出快速升高后趋于稳定。相关性分析表明,Saccharomycopsis与糖化酶、Aspergillus与蛋白酶、Wickerhamomyces与纤维素酶、滤纸酶等指标之间呈显著正相关性,Alternaria与糖化酶指标之间呈显著负相关性。优势菌株Saccharomycopsis、Aspergillus、Wickerhamomyces是影响六神曲发酵过程中酶学性质的主要因素。结论:该研究为六神曲微生物群落及多酶体系代谢研究提供了数据参考,为改良六神曲发酵工艺及指导规范化生产提供了理论依据。OBJECTIVE To explore the correlation between enzymatic properties and microbial colonial changes during fermentation with Massa Medicata Fermentata as a research object.METHODS The enzymatic characteristics of Massa Medicata Fermentata were evaluated by enzymatic activity detection.MiSeq high-throughput sequencing was utilized for examining microbial groups.And correlation analysis was performed for analyzing the correlation between the changes of enzyme properties and microbial colonies during fermentation.RESULTS During an initial stage of fermentation,microorganisms proliferated and such activity indices as amylase,glucoamylase,protease,lipase,filter paper enzyme and cellulase spiked sharply.In middle/late stage of fermentation,microbial species diminished and Aspergillus dominated.The peak glucoamylase activity reached 53.818μmol·g^(-1) at 36 h,amylase 87.841μmol·g^(-1) at 30 h and lipase,filter paper enzyme and cellulase 0.048,1.623 and 1.521μmol·g^(-1) at 24 h respectively,showing an initial rising trend and a subsequent decline.After 42 h,protease dropped sharply and then stabilized.Correlation analysis showed that significant positive correlations existed between Saccharomycopsis and glucoamylase,Aspergillus and protease,Wickerhamomyces and cellulase,filter paper enzyme and other indicators while a significant negative correlation existed between Alternaria and glucoamylase indicators.Dominant strains of Saccharomycopsis,Aspergillus and Wicker hamomyces were the major factors affecting the enzymatic properties of Massa Medicata Fermentata.CONCLUSION This study provides data supports for exploring microbial colonies and multienzyme system metabolism of Massa Medicata Fermentata.And theoretical rationales are offered for improving the fermentation technology of Massa Medicata Fermentata and guiding standardized production.

关 键 词：六神曲 发酵过程 酶学性质 微生物 相关分析 

分 类 号：R283.6[医药卫生—中药学] R966[医药卫生—中医学]