An efficient ABC transporter signal peptide directs heterologous protein secretion in food-grade hosts
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(2020) 36:154
ORIGINAL PAPER
An efficient ABC transporter signal peptide directs heterologous protein secretion in food‑grade hosts Ruangurai Promchai1 · Wonnop Visessanguan1 · Plearnpis Luxananil1 Received: 10 July 2020 / Accepted: 14 September 2020 © Springer Nature B.V. 2020
Abstract An efficient expression-secretion system for heterologous protein production in food-grade hosts, Lactobacillus plantarum and Bacillus subtilis, is still required to broaden their applications. The optimal signal peptide compatible with both the desired protein and the target host is important for the system. Here, we constructed new expression-secretion vectors to be used in both bacteria. A natural plasmid originating from food-grade L. plantarum BCC9546 was used as a core vector combined with a strong constitutive promoter, L-ldh promoter, and various signal peptides from several types of L. plantarum proteins: ABC transporter, cell wall-associated and extracellular proteins. A gene encoding 88-kDa amylase isolated from starch-related L. plantarum TBRC470 was used as a gene model to evaluate the systems. By comparing the amounts of secreted amylase from the recombinant strains to that of wild type, all signal peptides gave higher yields of secreted amylase in recombinant B. subtilis. Interestingly, two ABC transporter signal peptides from glutamine and mannose ABC transporters provided noticeably high levels of secreted amylase in recombinant L. plantarum. Moreover, these signal peptides also gave high yields of secreted amylase in recombinant B. subtilis. From the results, the signal peptide of glutamine ABC transporter, which functions in essential amino acid transportation that is a precursor for synthesis of nitrogen-containing compounds and nitrogen homeostasis, has a potential use in development of an efficient expression-secretion system for heterologous protein production in both food-grade hosts. Keywords Signal peptide · ABC transporter protein · Secretory vector · Lactobacillus plantarum · Bacillus subtilis
Introduction At present, recombinant DNA technology is highly useful for production of several enzymes and high-value substances. Gram-positive bacteria carrying the generally recognized as safe (GRAS) status such as Lactobacillus plantarum and Bacillus subtilis are attractive to be used as hosts for production of heterologous proteins in various industries including food and nutraceuticals (Landete 2017; Cui et al. 2018; Börner et al. 2019; Plavec and Berlec 2019). These bacteria are a good host for protein secretion since they possess a single membrane. Secretory protein production has several * Plearnpis Luxananil [email protected] 1
Microbial Cell Factory Research Team, National Center for Genetic Engineering and Biotechnology, National Science and Technology Development Agency, 113 Thailand Science Park, Phahonyothin Road, Klong 1, Klong Luang 12120, Pathumthani, Thailand
advantages such as eliminating all toxic effects of target protein accumulation in the cells, simplifying downstream processing
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