一株高产3-羟基丁酮枯草芽孢杆菌的构建

以产3-羟基丁酮枯草芽孢杆菌(Bacillus subtilis)TH-49(Val-)和产α-乙酰乳酸脱羧酶地衣芽孢杆菌(Bacillus licheniformis)AD-30为亲本菌株，以聚乙二醇为融合促进剂，进行原生质体融合获得融合子，融合子经再生、筛选等过程，最终获得高产3-羟基丁酮菌株HB-32，该菌株3-羟基丁酮产量高达49.64 g/L，比菌株TH-49(Val-)提高了61.8%，且遗传稳定性良好。进一步采用随机扩增多态性DNA(RAPD)技术，从分子水平上分析了高产3-羟基丁酮菌株HB-32与亲本菌株基因组的变化。结果表明，枯草芽孢杆菌(B. subtilis)TH-49(Val-)和地衣芽孢杆菌(B. licheniformis)AD-30原生质体融合成功，提高了融合子HB-32 3-羟基丁酮产量。

Construction of a Bacillus subtilis with high yield acetoin

Using polyethylene glycol as fusogen, the protoplasts of acetoin-producing Bacillus subtilis TH-49(Val-) and α-acetolactate decarboxylase-producing Bacillus licheniformis AD-30 were fused. By regeneration and screening of fusants, the fusant HB-32 with high acetoin production was obtained, and the yield of acetoin was up to 49.64 g/L, which was 61.8% higher than that of the strain TH-49(Val-). The fusant HB-32 had good genetic stability. The genome changes of fusant HB-32 and parent strains were analyzed in molecular level by random amplified polymorphic DNA. The results showed that the protoplast fusants of B. subtilis TH-49(Val-) and B. licheniformis AD-30 were fused successfully, and the yield of acetoin produced by fusant HB-32 was increased.