重组大肠杆菌制备珍珠贝源抗氧化肽发酵条件优化

Study on optimization of fermentation conditions for preparing antioxidant peptides of Pinctada fucata by recombinant E.coil

为进一步提高实验室前期构建的产抗氧化肽重组工程菌株DE3-p EGX6p-1-PFMAP制备珍珠贝源抗氧化肽的制备效率,通过摇瓶发酵过程中发酵条件包括诱导温度、诱导时间、IPTG浓度、培养基、摇床转速和接种量的单因素优化实验确定最佳发酵条件,并进一步利用正交实验优化50 L发酵罐发酵条件,结果表明摇瓶和50 L发酵罐最佳发酵条件一致,均为诱导温度30℃、诱导时间4 h、IPTG浓度0.2 mmol/L、培养基p H7、摇床转速220 r/min和接种量5%(v/v)。该工艺操作方便、过程简单、时间短,能高效制备出目的蛋白,为珠母贝源抗氧化肽的下一步工业化应用提供了有力技术支撑。

In order to improve the efficiency of preparing antioxidant peptides of Pinctada fucata by recombinant E. coil（DE3-pEGX6p-1-PFMAP） ,fermentation factors including induction temperature, induction time, concentration of IPTG, culture medium pH, shaker speed and inoculum size were optimized respectively in shake flask. Furthermore,the orthogonal experiment for the pilot- scale was optimized. Results showed that the best fermentation conditions of both shaking flask and orthogonal experiment, including induction temperature, induction time, concentration of IPTG, culture medium pH, shaker speed and inoculum size were 30 ℃,4 h ,0.2 mmol/L, pHT, 220 r/min,5% （v/v）respectively.And the process in this study was easy to operate and take a shorter time,which help further industrial application for preparing the antioxidant peptides of Pinctada fucata by recombinant Ecoil.