State Key Laboratory of Heavy Oil Processing and Centre for Bioengineering & Biotechnology, China University of Petroleum, Qingdao, 266580, PR China.
Biotechnol Biofuels. 2012 Dec 6;5(1):88. doi: 10.1186/1754-6834-5-88.
Acetoin and 2,3-butanediol are two important biorefinery platform chemicals. They are currently fermented below 40°C using mesophilic strains, but the processes often suffer from bacterial contamination.
This work reports the isolation and identification of a novel aerobic Geobacillus strain XT15 capable of producing both of these chemicals under elevated temperatures, thus reducing the risk of bacterial contamination. The optimum growth temperature was found to be between 45 and 55°C and the medium initial pH to be 8.0. In addition to glucose, galactose, mannitol, arabionose, and xylose were all acceptable substrates, enabling the potential use of cellulosic biomass as the feedstock. XT15 preferred organic nitrogen sources including corn steep liquor powder, a cheap by-product from corn wet-milling. At 55°C, 7.7 g/L of acetoin and 14.5 g/L of 2,3-butanediol could be obtained using corn steep liquor powder as a nitrogen source. Thirteen volatile products from the cultivation broth of XT15 were identified by gas chromatography-mass spectrometry. Acetoin, 2,3-butanediol, and their derivatives including a novel metabolite 2,3-dihydroxy-3-methylheptan-4-one, accounted for a total of about 96% of all the volatile products. In contrast, organic acids and other products were minor by-products. α-Acetolactate decarboxylase and acetoin:2,6-dichlorophenolindophenol oxidoreductase in XT15, the two key enzymes in acetoin metabolic pathway, were found to be both moderately thermophilic with the identical optimum temperature of 45°C.
Geobacillus sp. XT15 is the first naturally occurring thermophile excreting acetoin and/or 2,3-butanediol. This work has demonstrated the attractive prospect of developing it as an industrial strain in the thermophilic fermentation of acetoin and 2,3-butanediol with improved anti-contamination performance. The novel metabolites and enzymes identified in XT15 also indicated its strong promise as a precious biological resource. Thermophilic fermentation also offers great prospect for improving its yields and efficiencies. This remains a core aim for future work.
乙酰丙酮和 2,3-丁二醇是两种重要的生物炼制平台化学品。它们目前使用嗜温菌株在 40°C 以下发酵,但这些过程常常受到细菌污染的困扰。
本工作报道了一株新型好氧地芽孢杆菌(Geobacillus)菌株 XT15 的分离和鉴定,该菌株能够在高温下同时生产这两种化学品,从而降低细菌污染的风险。最佳生长温度为 45 至 55°C,培养基初始 pH 值为 8.0。除葡萄糖外,半乳糖、甘露醇、阿拉伯糖和木糖也都是可接受的底物,这使得纤维素生物质作为原料成为可能。XT15 优先使用有机氮源,包括玉米浆粉,这是玉米湿法加工的一种廉价副产品。在 55°C 下,以玉米浆粉为氮源可获得 7.7 g/L 的乙酰丙酮和 14.5 g/L 的 2,3-丁二醇。通过气相色谱-质谱联用仪鉴定了 XT15 培养液中的 13 种挥发性产物。乙酰丙酮、2,3-丁二醇及其衍生物(包括一种新型代谢物 2,3-二羟基-3-甲基庚烷-4-酮)占所有挥发性产物的 96%左右。相比之下,有机酸和其他产物则是次要副产物。XT15 中的两种关键酶,即α-乙酰乳酸脱羧酶和乙酰丙酮:2,6-二氯酚靛酚氧化还原酶,都是中度嗜热的,最佳温度相同,为 45°C。
地芽孢杆菌 XT15 是第一种自然产生的能分泌乙酰丙酮和/或 2,3-丁二醇的嗜热菌。本工作证明了将其开发为高温发酵生产乙酰丙酮和 2,3-丁二醇的工业菌株的诱人前景,具有提高抗污染性能的潜力。XT15 中鉴定的新型代谢物和酶也表明其作为一种宝贵的生物资源具有很大的潜力。高温发酵也为提高其产量和效率提供了广阔的前景。这仍然是未来工作的核心目标。