Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, No.189 Songling Road, Laoshan District, Qingdao, Shandong Province, 266101, China.
Qingdao Engineering Laboratory of Single Cell Oil, Qingdao, Shandong, 266101, China.
BMC Biotechnol. 2017 Aug 30;17(1):68. doi: 10.1186/s12896-017-0388-8.
Arachidonic acid (ARA, C20:4, n-6), which belongs to the omega-6 series of polyunsaturated fatty acids and has a variety of biological activities, is commercially produced in Mortierella alpina. Dissolved oxygen or oxygen utilization efficiency is a critical factor for Mortierella alpina growth and arachidonic acid production in large-scale fermentation. Overexpression of the Vitreoscilla hemoglobin gene is thought to significantly increase the oxygen utilization efficiency of the cells.
An optimized Vitreoscilla hemoglobin (VHb) gene was introduced into Mortierella alpina via Agrobacterium tumefaciens-mediated transformation. Compared with the parent strain, the VHb-expressing strain, termed VHb-20, grew faster under both limiting and non-limiting oxygen conditions and exhibited dramatic changes in cell morphology. Furthermore, VHb-20 produced 4- and 8-fold higher total lipid and ARA yields than those of the wild-type strain under a microaerobic environment. Furthermore, ARA production of VHb-20 was also 1.6-fold higher than that of the wild type under normal conditions. The results demonstrated that DO utilization was significantly increased by expressing the VHb gene in Mortierella alpina.
The expression of VHb enhances ARA and lipid production under both lower and normal dissolved oxygen conditions. This study provides a novel strategy and an engineered strain for the cost-efficient production of ARA.
花生四烯酸(ARA,C20:4,n-6)属于ω-6 系列多不饱和脂肪酸,具有多种生物学活性,在高山被孢霉中商业化生产。溶解氧或氧利用率是高山被孢霉大规模发酵生长和产生花生四烯酸的关键因素。过表达威氏血红蛋白基因被认为可以显著提高细胞的氧利用率。
通过根癌农杆菌介导的转化将优化的威氏血红蛋白(VHb)基因导入高山被孢霉。与亲本菌株相比,表达 VHb 的菌株(称为 VHb-20)在限氧和非限氧条件下生长更快,细胞形态发生显著变化。此外,在微需氧环境下,VHb-20 的总脂和 ARA 产量分别比野生型菌株高出 4 倍和 8 倍。此外,在正常条件下,VHb-20 的 ARA 产量也比野生型高出 1.6 倍。结果表明,在高山被孢霉中表达 VHb 基因可显著提高 DO 的利用率。
在较低和正常溶解氧条件下,表达 VHb 均可增强 ARA 和脂质的生产。本研究为高效生产 ARA 提供了一种新的策略和工程菌株。