Enhanced secretion of heterologous cyclodextrin glycosyltransferase by a mutant of Bacillus licheniformis defective in the D-alanylation of teichoic acids

Craynest, M.; Jørgensen, Steen T.; Sarvas, Matti; Kontinen, Vesa P.

doi:10.1046/j.1472-765x.2003.01357.x

Cited by 16 publications

(9 citation statements)

References 20 publications

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“…The B. subtilis dlt mutant had increased expression of heterologous amylase Q from B. amyloliquefaciens and the protective antigen from B. anthracis (14,37). Similar results for expression of other heterologous proteins were obtained in dlt mutants of Bacillus licheniformis and L. lactis (7,29). Thus, our results obtained with the dlt mutant are consistent with the literature.…”

Section: Discussionsupporting

confidence: 82%

Role of the Cell Wall Microenvironment in Expression of a Heterologous SpaP-S1 Fusion Protein by Streptococcus gordonii

Davis

Kennedy

Halperin

et al. 2011

Appl Environ Microbiol

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The charge density in the cell wall microenvironment of Gram-positive bacteria is believed to influence the expression of heterologous proteins. To test this, the expression of a SpaP-S1 fusion protein, consisting of the surface protein SpaP of Streptococcus mutans and a pertussis toxin S1 fragment, was studied in the live vaccine candidate bacterium Streptococcus gordonii. Results showed that the parent strain PM14 expressed very low levels of SpaP-S1. By comparison, the dlt mutant strain, which has a mutation in the dlt operon preventing D-alanylation of the cell wall lipoteichoic acids, and another mutant strain, OB219(pPM14), which lacks the LPXTG major surface proteins SspA and SspB, expressed more SpaP-S1 than the parent. Both the dlt mutant and the OB219(pPM14) strain had a more negatively charged cell surface than PM14, suggesting that the negative charged cell wall played a role in the increase in SpaP-S1 production. Accordingly, the addition of Ca 2؉ , Mg 2؉ , and K ؉ , presumably increasing the positive charge of the cell wall, led to a reduction in SpaP-S1 production, while the addition of bicarbonate resulted in an increase in SpaP-S1 production. The level of SpaP-S1 production could be correlated with the level of PrsA, a peptidyl-prolyl cis/trans isomerase, in the cells. PrsA expression appears to be regulated by the cell envelope stress two-component regulatory system LiaSR. The results collectively indicate that the charge density of the cell wall microenvironment can modulate heterologous SpaP-S1 protein expression in S. gordonii and that this modulation is mediated by the level of PrsA, whose expression is regulated by the LiaSR two-component regulatory system.

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Section: Discussionsupporting

confidence: 82%

Role of the Cell Wall Microenvironment in Expression of a Heterologous SpaP-S1 Fusion Protein by Streptococcus gordonii

Davis

Kennedy

Halperin

et al. 2011

Appl Environ Microbiol

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“…It was not clear from any of the dlt mutants previously described in the literature how the lack of D-alanylation of LTA might cause the expression of a surface protein to be lowered to such an extent. However, mutations in the dlt operon have been shown to result in perturbations at the cell membrane/cell wall interface and this has been linked to changes in post-transcriptional folding, degradation, and secretion of various proteins [8], [11], [14], [40]. To determine whether decreased M protein on the cell surface might be due to impaired retention, we examined M1 protein levels in the culture supernatant.…”

Section: Resultsmentioning

confidence: 99%

Inactivation of DltA Modulates Virulence Factor Expression in Streptococcus pyogenes

et al. 2009

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BackgroundD-alanylated lipoteichoic acid is a virtually ubiquitous component of Gram-positive cell walls. Mutations in the dltABCD operon of numerous species exhibit pleiotropic effects, including reduced virulence, which has been attributed to increased binding of cationic antimicrobial peptides to the more negatively charged cell surface. In this study, we have further investigated the effects that mutating dltA has on virulence factor expression in Streptococcus pyogenes.Methodology/Principal FindingsIsogenic ΔdltA mutants had previously been created in two distinct M1T1 isolates of S. pyogenes. Immunoblots, flow cytometry, and immunofluorescence were used to quantitate M protein levels in these strains, as well as to assess their ability to bind complement. Bacteria were tested for their ability to interact with human PMN and to grow in whole human blood. Message levels for emm, sic, and various regulatory elements were assessed by quantitative RT-PCR. Cell walls of ΔdltA mutants contained much less M protein than cell walls of parent strains and this correlated with reduced levels of emm transcripts, increased deposition of complement, increased association of bacteria with polymorphonuclear leukocytes, and reduced bacterial growth in whole human blood. Transcription of at least one other gene of the mga regulon, sic, which encodes a protein that inactivates antimicrobial peptides, was also dramatically reduced in ΔdltA mutants. Concomitantly, ccpA and rofA were unaffected, while rgg and arcA were up-regulated.Conclusions/SignificanceThis study has identified a novel mechanism for the reduced virulence of dltA mutants of Streptococcus pyogenes in which gene regulatory networks somehow sense and respond to the loss of DltA and lack of D-alanine esterification of lipoteichoic acid. The mechanism remains to be determined, but the data indicate that the status of D-alanine-lipoteichoic acid can significantly influence the expression of at least some streptococcal virulence factors and provide further impetus to targeting the dlt operon of Gram-positive pathogens in the search for novel antimicrobial compounds.

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“…), and the yields of cyclodextrin glycosyltransferase (Craynest et al . ), rPA (Thwaite et al . ), α‐amylase, nattokinase and β‐mannanase (Chen et al .…”

Section: Optimizing Secretion Pathwaymentioning

confidence: 99%

EngineeringBacillusfor efficient production of heterologous protein: current progress, challenge and prospect

et al. 2019

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Summary Bacillus species are widely recognized as good industrial platform strains, which can produce a variety of valuable products including enzymes and functional proteins. Bacillus expression systems gain various competitive edges over other expression systems, with respect to nontoxicity, convenience for gene modification and high yield of target proteins. Recently, a number of Bacillus expression systems have been developed, and various strategies were conducted to improve the yields of target proteins. In this review, we focused on the strategies of host strain optimization for heterologous protein production, including secretion pathway optimization, RNA and protein stability enhancement, cell growth facilitation, genome streamlining and transcriptional regulator engineering. In addition, Bacillus spore surface display and food‐grade expression systems were developed to expand the application of Bacillus expression system in recent years. Finally, the challenges and prospects of Bacillus expression system were discussed regarding the recent progresses, challenges and trends in this field.

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Enhanced secretion of heterologous cyclodextrin glycosyltransferase by a mutant of Bacillus licheniformis defective in the D-alanylation of teichoic acids

Cited by 16 publications

References 20 publications

Role of the Cell Wall Microenvironment in Expression of a Heterologous SpaP-S1 Fusion Protein by Streptococcus gordonii

Role of the Cell Wall Microenvironment in Expression of a Heterologous SpaP-S1 Fusion Protein by Streptococcus gordonii

Inactivation of DltA Modulates Virulence Factor Expression in Streptococcus pyogenes

EngineeringBacillusfor efficient production of heterologous protein: current progress, challenge and prospect

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