Ding Qiang, Liu Yadi, Hu Guipeng, Guo Liang, Gao Cong, Chen Xiulai, Chen Wei, Chen Jian, Liu Liming
State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, China.
International Joint Laboratory On Food Safety, Jiangnan University, Wuxi, 214122, China.
Bioresour Bioprocess. 2021 Nov 30;8(1):118. doi: 10.1186/s40643-021-00470-7.
Microbial organelles are a promising model to promote cellular functions for the production of high-value chemicals. However, the concentrations of enzymes and nanoparticles are limited by the contact surface in single Escherichia coli cells. Herein, the definition of contact surface is to improve the amylase and CdS nanoparticles concentration for enhancing the substrate starch and cofactor NADH utilization. In this study, two biofilm-based strategies were developed to improve the contact surface for the production of shikimate and L-malate. First, the contact surface of E. coli was improved by amylase self-assembly with a blue light-inducible biofilm-based SpyTag/SpyCatcher system. This system increased the glucose concentration by 20.7% and the starch-based shikimate titer to 50.96 g L, which showed the highest titer with starch as substrate. Then, the contact surface of E. coli was improved using a biofilm-based CdS-biohybrid system by light-driven system, which improved the NADH concentration by 83.3% and increased the NADH-dependent L-malate titer to 45.93 g L. Thus, the biofilm-based strategies can regulate cellular functions to increase the efficiency of microbial cell factories based on the optogenetics, light-driven, and metabolic engineering.
微生物细胞器是促进细胞功能以生产高价值化学品的一个有前景的模型。然而，单个大肠杆菌细胞中的酶和纳米颗粒浓度受接触表面的限制。在此，接触表面的定义是提高淀粉酶和硫化镉纳米颗粒的浓度，以增强底物淀粉和辅因子烟酰胺腺嘌呤二核苷酸（NADH）的利用。在本研究中，开发了两种基于生物膜的策略来改善用于生产莽草酸和L - 苹果酸的接触表面。首先，通过基于蓝光诱导生物膜的SpyTag/SpyCatcher系统进行淀粉酶自组装，改善了大肠杆菌的接触表面。该系统使葡萄糖浓度提高了20.7%，以淀粉为底物的莽草酸滴度达到50.96 g/L，这是使用淀粉作为底物时的最高滴度。然后，通过基于光驱动系统的生物膜硫化镉生物杂交系统改善了大肠杆菌的接触表面，该系统使NADH浓度提高了83.3%，并将依赖NADH的L - 苹果酸滴度提高到45.93 g/L。因此，基于生物膜的策略可以基于光遗传学、光驱动和代谢工程来调节细胞功能，以提高微生物细胞工厂的效率。
