Wu Hui, San Ka-Yiu
Department of Bioengineering, Rice University, Houston, Texas.
Biotechnol Bioeng. 2014 Nov;111(11):2209-19. doi: 10.1002/bit.25296. Epub 2014 Jul 14.
Free fatty acids (FFAs) can be used as precursors for the production of biofuels or chemicals. Different composition of FFAs will be useful for further modification of the biofuel/biochemical quality. Microbial biosynthesis of even chain FFAs can be achieved by introducing an acyl-acyl carrier protein thioesterase gene into E. coli. In this study, odd straight medium chain FFAs production was investigated by using metabolic engineered E. coli carrying acyl-ACP thioesterase (TE, Ricinus communis), propionyl-CoA synthase (Salmonella enterica), and β-ketoacyl-acyl carrier protein synthase III (four different sources) with supplement of extracellular propionate. By using these metabolically engineered E. coli, significant quantity of C13 and C15 odd straight-chain FFAs could be produced from glucose and propionate. The highest concentration of total odd straight chain FFAs attained was 1205 mg/L by the strain HWK201 (pXZ18, pBHE2), and 85% of the odd straight chain FFAs was C15. However, the highest percentage of odd straight chain FFAs was achieved by the strain HWK201 (pXZ18, pBHE3) of 83.2% at 48 h. This strategy was also applied successfully in strains carrying different TE, such as the medium length acyl-ACP thioesterase gene from Umbellularia californica. C11 and C13 became the major odd straight-chain FFAs.
游离脂肪酸(FFAs)可用作生产生物燃料或化学品的前体。不同组成的游离脂肪酸将有助于进一步改善生物燃料/生物化学物质的质量。通过将酰基-酰基载体蛋白硫酯酶基因导入大肠杆菌,可以实现偶数链游离脂肪酸的微生物生物合成。在本研究中,使用携带酰基-ACP硫酯酶(TE,蓖麻)、丙酰辅酶A合酶(肠炎沙门氏菌)和β-酮酰基-酰基载体蛋白合酶III(四种不同来源)并补充细胞外丙酸的代谢工程大肠杆菌,研究奇数直链中链游离脂肪酸的生产。通过使用这些代谢工程大肠杆菌,可以从葡萄糖和丙酸中产生大量的C13和C15奇数直链游离脂肪酸。菌株HWK201(pXZ18,pBHE2)获得的总奇数直链游离脂肪酸的最高浓度为1205 mg/L,其中85%的奇数直链游离脂肪酸为C15。然而,菌株HWK201(pXZ18,pBHE3)在48小时时达到了奇数直链游离脂肪酸的最高百分比,为83.2%。该策略也成功应用于携带不同硫酯酶的菌株,如来自加州月桂树的中链酰基-ACP硫酯酶基因。C11和C13成为主要的奇数直链游离脂肪酸。