Syngas Biofuels Energy, Inc., Houston, TX 77019, USA.
Lett Appl Microbiol. 2012 Aug;55(2):149-54. doi: 10.1111/j.1472-765X.2012.03272.x. Epub 2012 Jun 15.
To engineer acetogen biocatalyst capable of fermenting synthesis gas blend to acetone as the only liquid carbonaceous product.
The metabolic engineering comprised inactivation of phosphotransacetylase via integration of a cassette comprising synthetic genes erm(B), thiolase and HMG-CoA synthase. Acetaldehyde dehydrogenase was inactivated via integration of a cassette consisting of synthetic genes cat, HMG-CoA lyase and acetoacetate decarboxylase. The engineered biocatalyst Clostridum sp. MAceT113 lost production of 253 mmol l(-1) ethanol and 296 mmol l(-1) acetate and started producing 1.8 mol l(-1) acetone in single-stage continuous syngas fermentation.
The acetone concentration in culture broth is economical for bulk manufacture because it is about twenty times of that achieved with known acetone-butanol-ethanol fermentation of sugars.
The process shows the opportunity to produce acetone from synthesis gas at concentrations comparable with production of acetone from products of petroleum cracking. This is the first report on elimination of acetate and acetaldehyde production and directing carbon flux from Acetyl-CoA to acetone via a non-naturally occurring in acetogen acetone biosynthesis pathway identified in eukaryotic organisms.
构建能够利用合成气混合物发酵生产丙酮作为唯一液态碳源产物的乙酰辅酶 A 生物催化剂。
代谢工程通过整合包含合成基因 erm(B)、硫解酶和 HMG-CoA 合酶的基因盒,实现了磷酸转乙酰酶的失活。通过整合包含合成基因 cat、HMG-CoA 裂解酶和乙酰乙酸脱羧酶的基因盒,实现了乙醛脱氢酶的失活。工程化生物催化剂 Clostridium sp. MAceT113 失去了生产 253mmol/L 乙醇和 296mmol/L 乙酸的能力,并在单级连续合成气发酵中开始生产 1.8mol/L 的丙酮。
发酵液中的丙酮浓度对于大规模生产具有经济性,因为它是利用已知的糖的丙酮丁醇乙醇发酵所达到的浓度的约二十倍。
该工艺展示了从合成气生产丙酮的机会,其浓度可与从石油裂化产物生产丙酮相媲美。这是第一个关于消除乙酸和乙醛生产,并通过在真核生物中鉴定的非天然存在的乙酰辅酶 A 到丙酮的丙酮生物合成途径将碳通量从乙酰辅酶 A 定向到丙酮的报告。