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Hypersecretion of vaccine antigen outer membrane lipoprotein A in Corynebacterium glutamicum through high-throughput based development process
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  • Manman Sun,
  • Xiong Gao,
  • Rodrigo Ledesma-Amaro,
  • An Li,
  • Rongbing Wang,
  • Jiangqi Nie,
  • Pei Zheng,
  • Yankun Yang,
  • zhonghu bai,
  • xiuxia liu
Manman Sun
Jiangnan University

Corresponding Author:sunmanman0927@163.com

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Xiong Gao
The Hong Kong University of Science and Technology
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Rodrigo Ledesma-Amaro
Department of Bioengineering
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An Li
Jiangnan University
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Rongbing Wang
University of Turku
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Jiangqi Nie
Jiangnan University
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Pei Zheng
Tecon Biology CO.Ltd
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Yankun Yang
Jiangnan University
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zhonghu bai
National Engineering Laboratory for Cereal Fermentauition Technology
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xiuxia liu
National Engineering Laboratory for Cereal Fermentauition Technology
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Abstract

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 4×1 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.