Hypersecretion of OmlA antigen in Corynebacterium glutamicum through high-throughput based development process
: Sun Manman, Gao Alex Xiong, Ledesma-Amaro Rodrigo, Li An, Wang Rongbin, Nie Jianqi, Zheng Pei, Yang Yankun, Bai Zhonghu, Liu Xiuxia
Publisher: SPRINGER
: 2022
: Applied Microbiology and Biotechnology
: APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
: APPL MICROBIOL BIOT
: 106
: 2953
: 2967
: 15
: 0175-7598
: 1432-0614
DOI: https://doi.org/10.1007/s00253-022-11918-x
: https://doi.org/10.1007/s00253-022-11918-x
Outer membrane lipoprotein A (OmlA) is a vaccine antigen against porcine contagious pleuropneumonia (PCP), a disease severely affecting the swine industry. Here, we aimed to systematically potentiate the secretory production of OmlA in Corynebacterium glutamicum (C. glutamicum), a widely used microorganism in the food industry, by establishing a holistic development process based on our high-throughput culture platform. The expression patterns, expression element combinations, medium composition, and induction conditions were comprehensively screened or optimized in microwell plates (MWPs), followed by fermentation parameter optimization in a 4 x 1 L parallel fermentation system (CUBER4). An unprecedented yield of 1.01 g/L OmlA was ultimately achieved in a 5-L bioreactor following the scaling-up strategy of fixed oxygen mass transfer coefficient (kLa), and the produced OmlA antigen showed well-protective immunity against Actinobacillus pleuropneumoniae challenge. This result provides a rapid and reliable pipeline to achieve the hyper-production of OmlA, and possibly other recombinant vaccines, in C. glutamicum.