高产四甲基吡嗪微生物的筛选、鉴定及发酵条件优化

Screening, identification and fermentation conditions optimization of high yield tetramethylpyrazine microorganism

该研究以高温大曲为材料,采用传统培养分离及高效液相色谱(HPLC)法分离筛选高产四甲基吡嗪的菌株,通过分子生物学技术对其进行菌种鉴定,并通过单因素试验和响应面法对其产四甲基吡嗪的发酵工艺进行优化。结果表明,分离筛选获得一株高产四甲基吡嗪的菌株,编号为1-13,经鉴定其为解淀粉芽孢杆菌(Bacillus amyloliquefaciens),其产四甲基吡嗪的最优发酵条件为初始pH值7.0、发酵温度39.0℃、铵盐添加量50.0 g/L(发酵第6天时添加)。在此优化条件下发酵10 d,四甲基吡嗪产量可达(166.19±2.79)mg/L,较优化前(112.26 mg/L)提高48%。

High yield tetramethylpyrazine strains were isolated and screened from high-temperature Daqu by traditional culture separation and HPLC,and identified by molecular biotechnology.In addition,the fermentation process for tetramethylpyrazine production was optimized by single factor tests and response surface method.The results showed that a high yield tetramethylpyrazine strain was isolated and screened,numbered as 1-13,and identified as Bacillus amyloliquefaciens.The optimal fermentation conditions for the strain 1-13 producing tetramethylpyrazine were initial pH 7.0,fermentation temperature 39.0℃and ammonium salt addition 50.0 g/L(adding on the 6th day of fermentation).Under the optimal conditions,the yield of tetramethylpyrazine was(166.19±2.79)mg/L after fermentation 10 d,which was 48%higher than that before optimization(112.26 mg/L).