A novel strategy for enhancing extracellular secretion of recombinant proteins in Escherichia coli |
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Authors: | Lingqia Su Chenhua Xu Ronald W. Woodard Jian Chen Jing Wu |
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Affiliation: | 1. State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Ave, Wuxi, Jiangsu, 214122, China 2. School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, 1800 Lihu Ave, Wuxi, Jiangsu, 214122, China 3. Department of Medicinal Chemistry, University of Michigan, Ann Arbor, MI, 48109, USA
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Abstract: | Secretion of cytoplasmic expressed proteins into culture medium has significant commercial advantages in large-scale production of proteins. Our previous study demonstrated that the membrane permeability of Escherichia coli could be significantly improved when Thermobifida fusca cutinase, without a signal peptide, was expressed in cytoplasm. This study investigated the extracellular production of other recombinant proteins, including both secretory and cytosolic proteins, with co-expression of cutinase. When the secretory enzymes, xylanase and α-amylase, were co-expressed with cutinase, the culture period was shortened by half, and the productivity was 7.9 and 2.0-fold to that of their individual control without co-expression, respectively. When the normally cytosolic proteins, xylose isomerase and trehalose synthase, were co-expressed with cutinase, more than half of the target proteins were “secreted” into the culture medium. Moreover, by using β-galactosidase to detect membrane leakage, the improved secretion of the above model proteins was confirmed not to be due to cell lysis. The study provides a novel strategy for enhancing extracellular secretion of recombinant proteins in E. coli. |
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