应用内源前体策略提高杆菌发酵制备TTMP的能力  

High-Yield Fermentative Preparation of Tetramethylpyrazine using An Endogenous Precursor Approach by Bacillus sp.

作　　者：朱兵峰[1] 徐岩 范文来[1] 吴群[1] 

机构地区：[1]江南大学食品科学与技术国家重点实验室工业生物技术教育部重点实验室生物工程学院,江苏无锡214122

出　　处：《酿酒科技》2010年第12期17-22,共6页Liquor-Making Science & Technology

基　　金：国家支撑计划(2007BAK36B00;2008BAI63B06);长江学者和创新团队发展计划(IRT0532);高等学校学科创新引智计划(111-2-06);国家自然科学基金(NSFCNo.31000806;20872050;);江南大学食品科学与技术国家重点实验室(SKLF-MB-200801)的资助

摘　　要：基于前体乙偶姻和四甲基吡嗪(TTMP)分子结构的相关性,建立了一种适合于风味化合物TTMP筛选的内源前体筛选策略;并应用该策略,从酱香型高温大曲中获得1株TTMP产生菌XZ1124,该菌株能利用葡萄糖代谢产生大量前体乙偶姻,从而有效促进了TTMP的合成;根据菌落、细胞形态特征和生理生化特性以及16S rDNA序列分析,将该菌株鉴定为枯草杆菌(Bacillus subtilis)。碳氮源以及培养条件的单因素优化结果表明,供氧条件和培养温度对TTMP合成的影响最为突出。在最优条件下,摇瓶发酵和发酵罐培养时前体乙偶姻的积累量>20 g/L,TTMP的合成量>4.08 g/L。上述结果有效证明了内源前体筛选策略用于筛选具有结构类似前体的风味功能菌;TTMP合成工艺具有生产强度高且能以廉价的农副产品(豆饼粉)作为底物,极具工业应用前景。A spore-forming Bacillus sp.was obtained from a high-temperature Chinese Maotai-flavor Daqu using an endogenous precursor screening strategy.The Bacillus sp.was capable of producing a high level of 2,3,5,6-tetramethylpyrazine（TTMP） via a precursor of 3-hydroxy-2-butanone（HB）.The strain was characterized as Bacillus subtilis according to the morphological,physiological and biochemical properties as well as partial 16S rRNA gene sequences.Different carbon and nitrogen sources,as well as fermentation conditions were investigated.Optimization showed that oxygen supply and fermentation temperature were the most important process parameters.More than 4.08 g/L of TTMP was achieved with a high level of endogenous precursor HB accumulation（more than 20 g/L） when applying the optimized medium and cultivation conditions in both flask and fermentor cultures.Our data demonstrated the effectiveness of endogenous precursor strategy in screening of microorganisms that produced flavor compounds with structure related precursors.Moreover,the high yield of TTMP and the cheap agro-industrial product（soybean meal） as substrate indicated potential for industrial application.

关 键 词：微生物 枯草杆菌 内源前体 发酵制备 3-羟基丁酮 TTMP 

分 类 号：TS261[轻工技术与工程—发酵工程]