Su Lingqia, Ji Dening, Tao Xiumei, Yu Lingang, Wu Jing, Xia Yongmei
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China.
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China.
J Biotechnol. 2017 Jan 20;242:92-100. doi: 10.1016/j.jbiotec.2016.12.009. Epub 2016 Dec 8.
In this study, a gene encoding a putative lipase from Fusarium oxysporum was optimized via codon optimization and expressed in Pichia pastoris KM71. The gene product was identified as a phospholipase B (PLB). The engineered P. pastoris was further cultured in a 3.6-L bioreactor. After optimization of the induction conditions, this system produced 6.6mgmL protein and 6503.8UmL PLB activity in the culture medium. Efficient expression of this PLB in P. pastoris should reduce the costs of production and application. The purified enzyme, with a specific activity of 1170Umg, was optimally active at pH 5.0 and 55°C. The results of a degumming experiment performed using the recombinant PLB showed that the phosphorus content of a test oil was decreased from 75.88ppm to 3.3ppm in 2h under optimal reaction conditions. This study provides a basis for the industrial use of F. oxysporum PLB in oil degumming applications.
在本研究中，通过密码子优化对尖孢镰刀菌中一个假定脂肪酶的编码基因进行了优化，并在毕赤酵母KM71中进行表达。该基因产物被鉴定为磷脂酶B（PLB）。对工程化毕赤酵母进一步在3.6升生物反应器中培养。在优化诱导条件后，该系统在培养基中产生了6.6mg/mL的蛋白质和6503.8U/mL的PLB活性。该PLB在毕赤酵母中的高效表达应能降低生产和应用成本。纯化后的酶比活性为1170U/mg，在pH 5.0和55°C时活性最佳。使用重组PLB进行的脱胶实验结果表明，在最佳反应条件下，测试油的磷含量在2小时内从75.88ppm降至3.3ppm。本研究为尖孢镰刀菌PLB在油脂脱胶应用中的工业应用提供了依据。
