Factors Influencing Recombinant Protein Secretion Efficiency in Gram-Positive Bacteria: Signal Peptide and Beyond

Peng, Chong; Shi, Chaoshuo; Cao, Xue; Liu, Yu; Liu, Fufeng; Lu, Fuping

doi:10.3389/fbioe.2019.00139

Cited by 52 publications

(37 citation statements)

References 76 publications

(106 reference statements)

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“…We made a similar observation with leptin (14). While incomplete cleavage may be overcome by testing different leader sequences or by optimizing the combination of the leader sequence and the N-terminal sequence of the mature protein, these are not trivial tasks (19). Alternative approaches may therefore be more efficient.…”

Section: Discussionmentioning

confidence: 56%

Secretion of Recombinant Interleukin-22 by Engineered Lactobacillus reuteri Reduces Fatty Liver Disease in a Mouse Model of Diet-Induced Obesity

Schueler

Stapleton

et al. 2020

mSphere

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The incidence of metabolic syndrome continues to rise globally. In mice, intravenous administration of interleukin-22 (IL-22) ameliorates various disease phenotypes associated with diet-induced metabolic syndrome. In patients, oral treatment is favored over intravenous treatment, but methodologies to deliver IL-22 via the oral route are nonexistent. The goal of this study was to assess to what extent engineered Lactobacillus reuteri secreting IL-22 could ameliorate nonalcoholic fatty liver disease. We used a mouse model of diet-induced obesity and assessed various markers of metabolic syndrome following treatment with L. reuteri and a recombinant derivative. Mice that received an 8-week treatment of wild-type probiotic gained less weight and had a smaller fat pad than the control group, but these phenotypes were not further enhanced by recombinant L. reuteri. However, L. reuteri secreting IL-22 significantly reduced liver weight and triglycerides at levels that exceeded those of the probiotic wild-type treatment group. Our findings are interesting in light of the observed phenotypes associated with reduced nonalcoholic liver disease, in humans the most prevalent chronic liver disease, following treatment of a next-generation probiotic that is administered orally. Once biological and environmental containment strategies are in place, therapeutic applications of recombinant Lactobacillus reuteri are on the horizon. IMPORTANCE In humans, nonalcoholic fatty liver disease (NAFLD) is the most prevalent liver disease due to the increased prevalence of obesity. While treatment of NAFLD is often geared toward lifestyle changes, such as diet and exercise, the use of dietary supplements such as probiotics is underinvestigated. Here, we report that probiotic Lactobacillus reuteri reduces fatty liver in a mouse model of diet-induced obesity. This phenotype was further enhanced upon delivery of recombinant interleukin-22 by engineered Lactobacillus reuteri. These observations pave the road to a better understanding of probiotic mechanisms driving the reduction of diet-induced steatosis and to development of next-generation probiotics for use in the clinic. Ultimately, these studies may lead to rational selection of (engineered) probiotics to ameliorate fatty liver disease.

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Section: Discussionmentioning

confidence: 56%

Secretion of Recombinant Interleukin-22 by Engineered Lactobacillus reuteri Reduces Fatty Liver Disease in a Mouse Model of Diet-Induced Obesity

Schueler

Stapleton

et al. 2020

mSphere

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“…Using a staphylococcal nuclease (NucA) and the truncated form of a α-amylase (AmyA) derived from L. amylovorus as reporter proteins, they identified several signal peptides that resulted in efficient secretion of the reported proteins. However, the performance of the signal peptides may vary depending on the protein that is secreted ( Mathiesen et al, 2009 ; Peng et al, 2019 ).…”

Section: Introductionmentioning

confidence: 99%

Efficient Secretion and Recombinant Production of a Lactobacillal α-amylase in Lactiplantibacillus plantarum WCFS1: Analysis and Comparison of the Secretion Using Different Signal Peptides

et al. 2021

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Lactic acid bacteria (LAB) have been used as starter cultures and producers of enzymes, antimicrobial peptides or metabolites that contribute to the flavor, texture and safety of food products. Lactiplantibacillus plantarum, one of the best-studied LAB, is considered as safe and effective cell factory for food applications. In this study, our aim was to use L. plantarum as the producer for high levels of a food-grade lactobacillal α-amylase, which has potential applications in food, fermentation and feed industries. The native form of an α-amylase (AmyL) from L. plantarum S21, an amylolytic LAB isolated from Thai fermented rice noodles, was expressed in L. plantarum WCFS1 using the pSIP expression system. The secretion of the α-amylase was driven by the native signal peptides of the α-amylases from L. plantarum S21 (SP_AmyL) and Lactobacillus amylovorus NRRL B-4549 (SP_AmyA), as well as by three Sec-type signal peptides derived from L. plantarum WCFS1; Lp_2145, Lp_3050, and Lp_0373. Among the tested signal peptides, Lp_2145 appears to be the best signal peptide giving the highest total and extracellular enzymatic activities of α-amylase AmyL from L. plantarum S21, which were 13.1 and 8.1 kU/L of fermentation, respectively. These yields were significantly higher than the expression and secretion in L. plantarum WCFS1 using the native signal peptide SP_AmyL, resulting in 6.2- and 5.4-fold increase in total and extracellular activities of AmyL, respectively. In terms of secretion efficiency, Lp_0373 was observed as the most efficient signal peptide among non-cognate signal peptides for the secretion of AmyL. Real-time reverse-transcriptase quantitative PCR (RT-qPCR) was used to estimate the mRNA levels of α-amylase transcript in each recombinant strain. Relative quantification by RT-qPCR indicated that the strain with the Lp_2145 signal peptide-containing construct had the highest mRNA levels and that the exchange of the signal peptide led to a change in the transcript level of the target gene.

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“…In addition, understanding the presence or absence of SPs in the genes of interest is critical for choosing the appropriate recombinant protein expression and purification systems, as the intracellular accumulation of secretory proteins and toxins may be toxic to the host cells. Indeed, the ability of SPs to translocate proteins has been utilised in recombinant protein expression systems for high quality and quantity results (Futatsumori-Sugai and Tsumoto, 2010; Cho et al ., 2019; Karyolaimos et al ., 2019; Peng et al ., 2019).…”

Section: Introductionmentioning

confidence: 99%

Razor: annotation of signal peptides from toxins

Bhandari

Gardner

Lim

2020

Preprint

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Motivation: Signal peptides are responsible for protein transport and secretion and are ubiquitous to all forms of life. The annotation of signal peptides is important for understanding protein translocation and toxin secretion, optimising recombinant protein expression, as well as for disease diagnosis and metagenomics. Results: Here we explore the features of these signal sequences across eukaryotes. We find that different kingdoms have their characteristic distributions of signal peptide residues. Additionally, the signal peptides of secretory toxins have common features across kingdoms. We leverage these subtleties to build Razor, a simple yet powerful tool for annotating signal peptides, which additionally predicts toxin- and fungal-specific signal peptides based on the first 23 N-terminal residues. Finally, we demonstrate the usability of Razor by scanning all reviewed sequences from UniProt. Indeed, Razor is able to identify toxins using their signal peptide sequences only. Strikingly, we discover that many defensive proteins across kingdoms harbour a toxin-like signal peptide; some of these defensive proteins have emerged through convergent evolution, e.g. defensin and defensin-like protein families, and phospholipase families. Availability and implementation: Razor is available as a web application (https://tisigner.com/razor) and a command-line tool (https://github.com/Gardner-BinfLab/Razor).

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Factors Influencing Recombinant Protein Secretion Efficiency in Gram-Positive Bacteria: Signal Peptide and Beyond

Cited by 52 publications

References 76 publications

Secretion of Recombinant Interleukin-22 by Engineered Lactobacillus reuteri Reduces Fatty Liver Disease in a Mouse Model of Diet-Induced Obesity

Secretion of Recombinant Interleukin-22 by Engineered Lactobacillus reuteri Reduces Fatty Liver Disease in a Mouse Model of Diet-Induced Obesity

Efficient Secretion and Recombinant Production of a Lactobacillal α-amylase in Lactiplantibacillus plantarum WCFS1: Analysis and Comparison of the Secretion Using Different Signal Peptides

Razor: annotation of signal peptides from toxins

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