Merits of secretion of heterologous proteins from industrial microorganisms

Quax, WJ

doi:10.1007/bf02898715

Cited by 26 publications

(17 citation statements)

References 9 publications

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“…However, even under the optimal induction condition, the level of SpaP-S1 produced was very low. Similar problems have been reported in Bacillus subtilis, where yields of native proteins can be as much as grams per liter (11,30) but the production of heterologous proteins is significantly lower (33).…”

supporting

confidence: 62%

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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supporting

confidence: 62%

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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“…Moreover, good fermentation technologies exist for various bacilli (36,40,41,263). Many proteins can be secreted to very high levels by B. subtilis, such as the ␣-amylase AmyQ from Bacillus amyloliquefaciens (1 to 3 g/liter) (204), protein A from Staphylococcus aureus (Ͼ1 g/liter) (91), and human interleukin-3 (227). Although not precisely documented in the scientific literature, about 10-fold-higher secretion levels can be reached in optimized industrial fermentation systems using Bacillus amyloliquefaciens or Bacillus licheniformis strains.…”

Section: Scope Of This Review-the Secretomementioning

confidence: 99%

Signal Peptide-Dependent Protein Transport in Bacillus subtilis : a Genome-Based Survey of the Secretome

Tjalsma

Bolhuis

Jongbloed

et al. 2000

Microbiol Mol Biol Rev

751

768

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SUMMARY One of the most salient features of Bacillus subtilis and related bacilli is their natural capacity to secrete a variety of proteins into their environment, frequently to high concentrations. This has led to the commercial exploitation of bacilli as major “cell factories” for secreted enzymes. The recent sequencing of the genome of B. subtilis has provided major new impulse for analysis of the molecular mechanisms underlying protein secretion by this organism. Most importantly, the genome sequence has allowed predictions about the composition of the secretome, which includes both the pathways for protein transport and the secreted proteins. The present survey of the secretome describes four distinct pathways for protein export from the cytoplasm and approximately 300 proteins with the potential to be exported. By far the largest number of exported proteins are predicted to follow the major “Sec” pathway for protein secretion. In contrast, the twin-arginine translocation “Tat” pathway, a type IV prepilin-like export pathway for competence development, and ATP-binding cassette transporters can be regarded as “special-purpose” pathways, through which only a few proteins are transported. The properties of distinct classes of amino-terminal signal peptides, directing proteins into the various protein transport pathways, as well as the major components of each pathway are discussed. The predictions and comparisons in this review pinpoint important differences as well as similarities between protein transport systems in B. subtilis and other well-studied organisms, such as Escherichia coli and the yeast Saccharomyces cerevisiae. Thus, they may serve as a lead for future research and applications.

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“…After extensive efforts, only 29% of the total recombinant xylanase was secreted by pET22Mxyl recombinant after 24 h of induction . An attempt was, therefore, made to subclone metagenomic xylanase gene into shuttle vector pWH1520 and express it in B. subtilis , since this has GRAS (Generally Recognized As Safe) status by Food and Drug Administration (FDA), USA and used frequently for the secretion of recombinant proteins because of its elaborate transcription and translation mechanisms …”

Section: Discussionmentioning

confidence: 99%

Production of cellulase‐free xylanase by the recombinant Bacillus subtilis and its applicability in paper pulp bleaching

Verma

Satyanarayana

2013

Biotechnology Progress

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A metagenomic xylanase gene (Mxyl) was successfully cloned into shuttle vector pWH1520 and expressed in Bacillus subtilis extracellularly. On induction with xylose, recombinant xylanase secretion commenced after 6 h. Identifying critical variables for recombinant xylanase production by one-variable-at-time approach followed by optimization of the selected variables (xylose, inoculum density, incubation density) by response surface methodology (RSM) led to three-fold enhancement in extracellular xylanase production (119 U mL(-1) ). When the pulp was treated with recombinant xylanase at 80°C and pH 9.0, kappa number of the pulp was reduced with concomitant increase in brightness and 24% reduction in chlorine consumption. This is the first report on the expression of metagenomic xylanase gene in Bacillus subtilis extracellularly and its utility in developing an environment-friendly pulp bleaching process.

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Merits of secretion of heterologous proteins from industrial microorganisms

Cited by 26 publications

References 9 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

Signal Peptide-Dependent Protein Transport in Bacillus subtilis : a Genome-Based Survey of the Secretome

Production of cellulase‐free xylanase by the recombinant Bacillus subtilis and its applicability in paper pulp bleaching

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