摘要
为了解市场上霉菌发酵食品能否扩增霉菌DNA,除了红曲霉外其他曲霉是否具有pksCT基因,以及加热灭菌条件对红曲黄酒中pksCT基因降解的影响。从市售的11种食品中提取DNA,选用ITS基因引物和pksCT基因引物进行聚合酶链式反应(polymerase chain reaction,PCR)扩增,并电泳分析。分别用麦曲、红曲发酵黄酒,对发酵前、发酵中、灭菌后、贮藏期样品扩增pksCT基因并测定橘霉素质量浓度;选择灭菌温度70、80、90、100℃,灭菌时间10、20、30 min,灭菌后样品扩增pksCT基因。结果表明有4种食品能用ITS引物扩增出条带,但只有红曲霉发酵的红腐乳能扩增出pksCT基因。红曲黄酒在发酵中、灭菌后能够扩增出pksCT基因,并且在80℃灭菌20 min和90℃灭菌10 min条件下,仍然可以扩增出pksCT基因,但90℃灭菌20 min或100℃灭菌10 min,DNA受到破坏。红曲霉发酵食品能够扩增出pksCT基因,曲霉发酵的食品未能用pksCT基因引物扩增出条带。通过PCR方法扩增出部分霉菌发酵食品的霉菌DNA并测序,有助于了解其加工中使用的菌种,继而为发酵加工食品的终端监测提供一种新方法。
This study aimed to find out whether mold DNA can be amplified from traditional mold fermented foods andwhether Aspergillus besides Monascus carries pksCT gene.The effect of thermal sterilization conditions on the degradationof the pksCT gene in red kojic rice wine was examined as well.Eleven samples of soy sauce,rice vinegar,fermented beancurd and yellow rice wine collected from local market were used for DNA extraction.Polymerase chain reaction(PCR)amplification was carried out with internal transcribed spacer(ITS)and pksCT gene primers.Rice wine was fermented bywheat koji and red koji,respectively,and then the pksCT gene was amplified and citrinin contents were measured beforeand during fermentation,after sterilization,and during storage.Samples were sterilized at70,80,90,100℃for10,20,and30min,respectively,and then were subjected to PCR amplification.The results showed that DNA fragments weresuccessfully amplified with the ITS primer from4samples,including soy sauce,rice vinegar,fermented bean curd andyellow rice wine.However,the pksCT gene was successfully amplified only from red bean curd fermented by Monascus.Wefailed to amplify the pksCT gene from yellow rice wine fermented by wheat koji.The pksCT gene was successfully amplifiedfrom red kojic rice wine during fermentation and after sterilization.Meanwhile,citrinin content increased rapidly at thesesampling points.When red kojic rice wine was sterilized at80℃for20min or90℃for10min,the pksCT gene remainedamplifiable,while after sterilization at90℃for20min or100℃for10min,DNA structure was damaged.The pksCTgene was amplifiable in Monascus fermented foods,but not in Aspergillus fermented foods.It is implied that mold DNAin fermented foods can be amplified by PCR and sequenced,which may be helpful for inspectors to find out what kinds of strains are contained in mold fermented foods,and provides a new terminal monitoring method for mold fermented foods.
作者
顾双
陈俊霖
王向阳
GU Shuang;CHEN Junlin;WANG Xiangyang(School of Food Science and Biotechnology, Zhejiang Gongshang University, Hangzhou 310018, China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2017年第4期83-86,共4页
Food Science
基金
浙江省自然科学基金项目(LY15C200005)
