黄酒发酵过程中乳酸菌的分离及对其产生物胺能力的评价  

Detection of biogenic amines produced by lactic acid bacteria isolated from the fermentation process of Chinese rice wine

作　　者：王然然[1,2,3] 李晓敏[1,2,3] 陈柳 卞小稳 蔡国林[1,2,3] 陆健[1,2,3] 

机构地区：[1]江南大学,工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学,粮食发酵工艺与技术国家工程实验室,江苏无锡214122 [3]江南大学生物工程学院,江苏无锡214122 [4]上海华润雪花啤酒(上海)有限公司,上海200949

出　　处：《食品与发酵工业》2017年第1期12-17,共6页Food and Fermentation Industries

基　　金：工业过程高效转化与系统集成的科学基础研究,973项目(2013CB733602);黄酒发酵过程中乳酸菌积累有害胺类物质代谢物的机制解析,江苏省自然科学基金(BK20140144);江苏高校优势学科建设工程资助项目

摘　　要：对黄酒酿造过程中乳酸菌进行分离,并对乳酸菌产生物胺能力进行检测。利用脱羧培养基对分离出的82株乳酸菌进行培养,结合反相高效液相色谱技术(reversed-phase high-performance liquid chromatography,RPHPLC)测定发酵液中生物胺含量,具体色谱条件如下:使用丹磺酰氯(Dansyl Chloride,Dns-CL)进行柱前衍生,以乙腈-乙腈(φ=50%)为流动相梯度洗脱,流速1.0 m L/min,254 nm紫外波检测。该方法在给定浓度范围内线性关系良好(R^2>0.996),平均回收率为94.46%~105.20%,相对偏差(RSD)均小于5%。检测到组胺产生菌株11株,最大生成量为18.19 mg/L,高产菌株分离自黄酒前酵第2天;酪胺产生菌株28株,最大生成量为30.38mg/L,高产菌株分离自黄酒前酵第2天;腐胺产生菌株51株,最大生成量为299.94 mg/L,高产菌株分离自浸米水。由此表明,有必要控制黄酒发酵过程中的乳酸菌,采用低产或不产生物胺的乳酸菌作为强化菌株,可降低黄酒中的生物胺含量。The performance of biogenic amine （BA） production which was conducted by lactic acid bacteria （LAB） isolated from the fomentation process of Chinese rice wine was explored in this paper. A collection of 82 LAB strains cultured in decarboxylation medium were screened for potential producers and BA content of each medium cul- ture was determined by reversed-phase high-performance liquid chromatography （RP-HPLC） after derivatized with dansyl chloride （Dns-cL）. The RP-HPLC analysis for the samples were conducted as follows： mobile phase was a gradient elution program of acetonitrile and acetonitrile （φ = 50% ） with a flow rate of 1 mL/min and the column ef- fluent was monitored at 254 nm with a UV detector. It turned out that the method for BA production presented good linearity （R2 〉0. 996） at given concentrations and a recovery rate of 94.46% -105.20% , together with a relative standard deviation （RSD） of less than 5%. Furthermore, there were 11 strains detected for histamine production （ maximum = 18.19 mg/L） and the max-histamine producing strain were isolated on the 2nd day of pre-fermentation. 9 strains for tyramine （maximum = 30.38 mg/L） and the max- tyramine producing strain were isolated on the 2nd day of pre-fermentation. 51 strains for putrescine （ maximum = 299.94 rag/L） and the max-putrescine producing strain were isolated from the rice immersion water. The results suggested that it was necessary to control LAB during Chinese rice wine fermentation to reduce the content of BA.

关 键 词：黄酒 乳酸菌 生物胺 反相高效液相色谱技术 

分 类 号：TS262.4[轻工技术与工程—发酵工程]