乙醇降解乳酸菌的筛选、鉴定及解酒防醉功效评价

Screening, identification, and anti-alcoholic and anti-drunken efficacy evaluation of ethanol-degrading lactic acid bacteria

该研究采用传统培养分离法从传统发酵食品奶疙瘩样品中分离乳酸菌,通过体外降解乙醇试验筛选高效降解乙醇功能的菌株,通过形态学观察、生理生化实验及分子生物学技术对其进行菌种鉴定,并对其进行安全性评价、耐逆性试验,以及体内小鼠防醉解酒功效试验。结果表明,筛选鉴定出一株乙醇降解率高(13.38%)、乙醇耐受能力强(耐受体积分数12%乙醇)的乳酸片球菌(Pediococcus acidilactici)RH2712,其对氨苄西林、青霉素、亚胺培南、氯霉素敏感,溶血性安全;对酸和胆盐耐受力强,在pH 3.0的条件下活菌对数值几乎不变,延迟时间为2 h,在1.5%胆盐含量下培养4 h后活菌对数值达到8.72,延迟时间仅为0.5 h,在人工模拟胃液条件下处理3 h后活菌对数值仍能达到7.14。乳酸片球菌RH2712在小鼠试验中表现出明显的防醉解酒的功效,且活菌数为1×10^(8)~1×10^(9)CFU/m L时解酒功能强于美他多辛。

Lactic acid bacteria were isolated from milk pimple(traditional fermented food)samples by traditional culture separation method,and the strains with high ethanol degradation function were screened by ethanol degradation test in vitro,and were identified by morphological observation,physiological and biochemical experiments and molecular biology techniques.Safety evaluation,stress tolerance tests,anti-alcoholic and anti-drunken efficacy tests in mice of screened strain were carried out.The results showed that a strain of Pediococcus acidilactici RH2712 with high ethanol degradation rate(13.38%)and strong ethanol tolerance(tolerant volume fraction 12%ethanol)was screened and identified.The strain was sensitive to antibiotics,such as ampicillin,penicillin,imipenem and chloromycetin,but with safe hemolysis.It had strong tolerance to acid and bile salt,and the logarithm value of living bacteria was almost unchanged under the condition of pH 3.0,and the delay time was 2 h.After cultured for 4 h under 1.5%bile salt content,the logarithm value of living bacteria reached 8.72,and the delay time was only 0.5 h.Under the condition of artificial simulated gastric juice,the logarithm value of living bacteria could reach 7.14 after 3 h.P.acidilactici RH2712 showed obvious anti-alcoholic and anti-drunken efficacy in mice,and the anti-alcoholic function was stronger than metadoxine when the number of living bacteria was 1×10^(8)-1×10^(9) CFU/ml.