School of Biotechnology, Institute of Agricultural Technology, Suranaree University of Technology, 111 University Ave., Suranaree, Muang, Nakhon Ratchasima 30000, Thailand.
Bioresour Technol. 2012 Jan;103(1):329-36. doi: 10.1016/j.biortech.2011.09.096. Epub 2011 Oct 7.
Sucrose-utilizing genes (cscKB and cscA) from Escherichia coli KO11 were cloned and expressed in a metabolically engineered E. coli KJ122 to enhance succinate production from sucrose. KJ122 harboring a recombinant plasmid, pKJSUC, was screened for the efficient sucrose utilization by growth-based selection and adaptation. KJ122-pKJSUC-24T efficiently utilized sucrose in a low-cost medium to produce high succinate concentration with less accumulation of by-products. Succinate concentrations of 51 g/L (productivity equal to 1.05 g/L/h) were produced from sucrose in anaerobic bottles, and concentrations of 47 g/L were produced in 10L bioreactor within 48 h. Antibiotics had no effect on the succinate production by KJ122-pKJSUC-24T. In addition, succinate concentrations of 62 g/L were produced from sugarcane molasses in anaerobic bottles, and concentrations of 56 g/L in 10 L bioreactor within 72 h. These results demonstrated that KJ122-pKJSUC-24T would be a potential strain for bio-based succinate production from sucrose and sugarcane molasses.
从大肠杆菌 KO11 中克隆并表达了蔗糖利用基因(cscKB 和 cscA),以提高大肠杆菌 KJ122 利用蔗糖生产琥珀酸的能力。通过基于生长的选择和适应,筛选出携带重组质粒 pKJSUC 的 KJ122,以实现高效利用蔗糖。KJ122-pKJSUC-24T 在低成本培养基中有效地利用蔗糖,产生高浓度的琥珀酸,同时副产物积累较少。在厌氧瓶中,KJ122-pKJSUC-24T 从蔗糖中生产出 51 g/L 的琥珀酸(相当于 1.05 g/L/h 的生产率),在 10L 生物反应器中 48 小时内生产出 47 g/L 的琥珀酸。抗生素对 KJ122-pKJSUC-24T 的琥珀酸生产没有影响。此外,在厌氧瓶中,KJ122-pKJSUC-24T 从甘蔗废糖蜜中生产出 62 g/L 的琥珀酸,在 10L 生物反应器中 72 小时内生产出 56 g/L 的琥珀酸。这些结果表明,KJ122-pKJSUC-24T 可能成为从蔗糖和甘蔗废糖蜜生产生物基琥珀酸的潜在菌株。