嘌呤核苷磷酸化酶的表达及其在啤酒中的应用  

Expression of Purine Nucleoside Phosphorylase and Its Application in Beer

作　　者：李玉淼 朱德伟 朱洪康[1,2,3] 孙军勇 陆健 LI Yumiao;ZHU Dewei;ZHU Hongkang;SUN Junyong;LU Jian(National Engineering Laboratory for Cereal Fermentation Technology,Jiangnan University,Wuxi 214122, China;Key Laboratory of Industrial Biotechnology,Ministry of Education,Jiangnan University,Wuxi 214122, China;School of Biotechnology,Jiangnan University,Wuxi 214122,China;Industrial Technology Research Institute of Jiangnan University in Suqian,Suqian 223800, China)

机构地区：[1]粮食发酵工艺与技术国家工程实验室,江南大学,江苏无锡214122 [2]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [3]江南大学生物工程学院,江苏无锡214122 [4]宿迁市江南大学产业技术研究院,江苏宿迁223800

出　　处：《食品与生物技术学报》2017年第12期1264-1268,共5页Journal of Food Science and Biotechnology

基　　金：国家863计划项目(2013AA102109);江苏省普通高等学校科研成果产业化推动项目(JHB2012-26);高等学校学科创新引智计划(111计划)项目(111-2-06);江苏高校优势学科建设工程项目

摘　　要：为了开发一种能水解核苷的嘌呤核苷磷酸化酶(Purine Nucleoside Phosphorylase,PNPase),将其添加到糖化醪液中,增加麦汁中的游离嘌呤碱基质量浓度,在后续发酵阶段游离嘌呤碱基被酵母利用,提高了嘌呤物质的利用率,最终啤酒中的嘌呤质量浓度减少。将PNPase的编码基因deo D导入表达载体pET-28a(+)中,采用化学转化法将载体转入到大肠杆菌BL21(DE3)进行诱导表达,研究重组PNPase的活性,并将其应用到糖化工艺中。获得的重组PNPase活性为184.46 U/mL,添加到糖化醪液中后,麦汁中游离嘌呤碱基质量浓度升高,酵母可吸收的游离嘌呤质量浓度增多,啤酒发酵液嘌呤质量浓度显著降低,为后续研究使啤酒嘌呤含量的降低奠定了一定的基础。To develop a purine nucleoside phosphorylase which can hydrolysis nucleoside. Adding purine nucleoside phosphorylase to the mash liquid, it could increase the free purine content in wort, which were utilized by yeast during the fermentation stage. As result,the utilization of purine could improve,purine content decreased in beer. The gene deoD encoding PNPase was cloned into the expression vector pET-28a （S） and the expression vector was transformed into E.coli BL21 （DE3） using chemical transformation. The activity of recombinant PNPase was analyzed and it was applied to the mashing process. The results showed that the enzyme activity of obtained recombinant PNPase was 184.46 U/mL. When it was added to the mash liquid, the free purine content in wort was increased. Thus, yeast could use more free purine, purine content decreased significantly in beer, laing the foundation for the further reduction of purine content in beer.

关 键 词：嘌呤核苷鱗酸化酶 原核表达 嘌呤碱基 啤酒 

分 类 号：O629.8[理学—有机化学] TS262.5[理学—化学]