产β-D-葡萄糖苷酶菌株筛选及其在香草兰豆荚发酵生香中的应用  

Isolation ofβ-D-glucosidase Producing Microorganisms and Application on Vanilla Pods Fermentation

作　　者：周烨 侯梅芳 赵青云 谢心湄[4] 徐飞 吴桂苹[3,5] 谷风林 ZHOU Ye;HOU Meifang;ZHAO Qingyun;XIE Xinmei;XU Fei;WU Guiping;GU Fenglin(School of Ecological Technology and Engineering,Shanghai Institute of Technology,Shanghai 200233,China;Sanya Research Institute,Chinese Academy of Tropical Agricultural Sciences,Sanya,Hainan 572000,China;Spice and Beverage Research Institute,Chinese Academy of Tropical Agricultural Sciences,Wanning,Hainan 571533,China;College of Food Science and Technology,Huazhong Agricultural University,Wuhan,Hubei 430070,China;Key Laboratory of Processing Suitability and Quality Control of the Special Tropical Crops of Hainan Province,Wanning,Hainan 571533,China)

机构地区：[1]上海应用技术大学生态技术工程学院,上海200233 [2]中国热带农业科学院三亚研究院,海南三亚572000 [3]中国热带农业科学院香料饮料研究所,海南万宁571533 [4]华中农业大学食品科学技术学院,湖北武汉430070 [5]海南省特色热带作物适宜性加工与品质控制重点实验室,海南万宁571533

出　　处：《热带作物学报》2024年第10期2232-2242,共11页Chinese Journal of Tropical Crops

基　　金：三亚崖州湾科技城科技专项(No.SCKJ-JYRC-2022-25);海南省重点研发计划项目(No.ZDYF2022XDNY268);中央级公益性科研院所基本科研业务费专项(No.1630142022008)。

摘　　要：本研究从发酵后的香草兰豆荚中筛选出产β-D-葡萄糖苷酶的微生物菌株,并将筛选出的菌株应用到香草兰发酵过程中,以提高发酵豆荚品质。采用七叶苷选择培养基传统分离培养方法,从发酵后的香草兰豆荚中筛选出可产β-D-葡萄糖苷酶的菌株,鉴定每株菌的菌株同源性,测量每株菌产β-D-葡萄糖苷酶酶活,绘制2株产酶较高的菌株系统发育树,并将产酶最高的2株菌按照不同组合喷洒至发酵中的香草兰豆荚中,测定其发酵豆荚香兰素含量、香兰素葡萄糖苷含量和挥发性成分。结果表明:从产自马达加斯加(M)的香草兰发酵豆荚中共筛选出9株菌、产自中国热带农业科学院香料饮料研究所(S)的香草兰发酵豆荚中筛选到12株菌,检测出的15株菌均为芽孢杆菌,其中M3、S7的β-D-葡萄糖苷酶活力较高。使用M3、S7以不同组合添加喷菌发酵试验的豆荚中,只喷M3的发酵豆荚香兰素含量最高为2.25%,M3与S7按照体积比1∶1喷洒处理的香兰葡萄糖苷含量下降最多,为80.7%,只喷M3的发酵豆荚鉴定出挥发性物质34种。本研究成功筛选出产β-D-葡萄糖苷酶的功能微生物,并利用筛选出的微生物辅助香草兰豆荚发酵生香,可用于提升香草兰豆荚品质。This study aims to screen the microbial strains capable of producingβ-D-glucosidase from fermented vanilla pods and apply the screened strains to the fermentation process of vanilla with the goal of enhancing vanillin production and improving the quality of vanilla bean.Septenary selective medium was utilized to isolateβ-D-glucosidase producing bacteria,identify the homology of each bacterial strain and measure the production ofβ-D-glucosidase activity of each strain on two strains with higher enzyme production.The phylogenetic tree was constructed.The two strains of bacteria with the highest enzyme production in different combinations were sprayed onto the fermented vanilla pods.The content of vanillin,vanillin glucoside,and volatile components in different treatments ofβ-D-glucosidase producing bacteria were determined.Nine strains were successfully screened from the vanilla pods originating in Madagascar(M),and twelve strains were isolated from the vanilla pods produced by Spice and Beverage Research Institute,Chinese Academy of Tropical Agricultural Sciences(S).The fifteen strains were detected as Bacillus.M3 and S7 had higher enzyme activities than other isolates.Vanilla pods treated with M3 obtained the highest vanillin content,which reached to 2.25%and possed the highest aroma components.The maximum decrease in vanillin glucoside content was 80.7%when sprayed with M3 and S7 in a volume ratio of 1:1.There were 34 volatile substances in the fermented pods sprayed with M3,making it the most volatile combination of spraying bacteria.In the present study,we successfully isolatedβ-D-glucosidase producing bacteria for improving the quality of vanilla pods fermentation.

关 键 词：香草兰 β-D-葡萄糖苷酶 香兰素 发酵优化 

分 类 号：Q815[生物学—生物工程]