Wang Dan, Thakker Chandresh, Liu Ping, Bennett George N, San Ka-Yiu
Department of Bioengineering, Rice University, 6100 Main Street, MS-362, Houston, Texas, 77005-1892.
College of Chemistry and Chemical Engineering, Chongqing University, Chongqing, P. R. China.
Biotechnol Bioeng. 2015 Nov;112(11):2324-33. doi: 10.1002/bit.25633. Epub 2015 Jul 31.
Conversion of biomass feedstock to chemicals and fuels has attracted increasing attention recently. Soybean meal, containing significant quantities of carbohydrates, is an inexpensive renewable feedstock. Glucose, galactose, and fructose can be obtained by enzymatic hydrolysis of soluble carbohydrates of soybean meal. Free fatty acids (FFAs) are valuable molecules that can be used as precursors for the production of fuels and other value-added chemicals. In this study, free fatty acids were produced by mutant Escherichia coli strains with plasmid pXZ18Z (carrying acyl-ACP thioesterase (TE) and (3R)-hydroxyacyl-ACP dehydratase) using individual sugars, sugar mixtures, and enzymatic hydrolyzed soybean meal extract. For individual sugar fermentations, strain ML211 (MG1655 fadD(-) fabR(-) )/pXZ18Z showed the best performance, which produced 4.22, 3.79, 3.49 g/L free fatty acids on glucose, fructose, and galactose, respectively. While the strain ML211/pXZ18Z performed the best with individual sugars, however, for sugar mixture fermentation, the triple mutant strain XZK211 (MG1655 fadD(-) fabR(-) ptsG(-) )/pXZ18Z with an additional deletion of ptsG encoding the glucose-specific transporter, functioned the best due to relieved catabolite repression. This strain produced approximately 3.18 g/L of fatty acids with a yield of 0.22 g fatty acids/g total sugar. Maximum free fatty acids production of 2.78 g/L with a high yield of 0.21 g/g was achieved using soybean meal extract hydrolysate. The results suggested that soybean meal carbohydrates after enzymatic treatment could serve as an inexpensive feedstock for the efficient production of free fatty acids.
生物质原料转化为化学品和燃料最近已引起越来越多的关注。豆粕含有大量碳水化合物,是一种廉价的可再生原料。葡萄糖、半乳糖和果糖可通过豆粕中可溶性碳水化合物的酶促水解获得。游离脂肪酸(FFA)是有价值的分子,可用作生产燃料和其他增值化学品的前体。在本研究中,携带质粒pXZ18Z(携带酰基-ACP硫酯酶(TE)和(3R)-羟基酰基-ACP脱水酶)的突变大肠杆菌菌株使用单糖、糖混合物和酶解豆粕提取物生产游离脂肪酸。对于单糖发酵,菌株ML211(MG1655 fadD(-) fabR(-) )/pXZ18Z表现最佳,在葡萄糖、果糖和半乳糖上分别产生4.22、3.79、3.49 g/L游离脂肪酸。虽然菌株ML211/pXZ18Z在单糖发酵中表现最佳,然而,对于糖混合物发酵,额外缺失编码葡萄糖特异性转运蛋白的ptsG的三突变菌株XZK211(MG1655 fadD(-) fabR(-) ptsG(-) )/pXZ18Z由于缓解了分解代谢物阻遏而表现最佳。该菌株产生约3.18 g/L脂肪酸,产率为0.22 g脂肪酸/g总糖。使用豆粕提取物水解物实现了最大游离脂肪酸产量2.78 g/L,高产率为0.21 g/g。结果表明,酶处理后的豆粕碳水化合物可作为高效生产游离脂肪酸的廉价原料。