<i>Bacillus subtilis</i> levansucrase: amino acid substitutions at one site affect secretion efficiency and refolding kinetics mediated by metals

Petit‐Glatron, Marie‐Françoise; Monteil, I.; Benyahia, Fadila; Chambert, Régis

doi:10.1111/j.1365-2958.1990.tb00566.x

Cited by 29 publications

(16 citation statements)

References 35 publications

(22 reference statements)

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“…On the other hand, these proteins might undergo rapid folding right after their synthesis, which interferes with (or hampers) the secretion process. Such interferences between protein conformation and SE were previously shown in E. coli and B. subtilis [32,33]. Altogether, these results suggest that protein conformation rather than protein size is a major problem for heterologous protein secretion in L. lactis as well.…”

Section: Protein Conformation Rather Than Protein Size Can Impair Thesupporting

confidence: 72%

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et al. 2005

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Lactococcus lactis, the model lactic acid bacterium (LAB), is a food grade and well-characterized Gram positive bacterium. It is a good candidate for heterologous protein delivery in foodstuff or in the digestive tract. L. lactis can also be used as a protein producer in fermentor. Many heterologous proteins have already been produced in L. lactis but only few reports allow comparing production yields for a given protein either produced intracellularly or secreted in the medium. Here, we review several works evaluating the influence of the localization on the production yields of several heterologous proteins produced in L. lactis. The questions of size limits, conformation, and proteolysis are addressed and discussed with regard to protein yields. These data show that i) secretion is preferable to cytoplasmic production; ii) secretion enhancement (by signal peptide and propeptide optimization) results in increased production yield; iii) protein conformation rather than protein size can impair secretion and thus alter production yields; and iv) fusion of a stable protein can stabilize labile proteins. The role of intracellular proteolysis on heterologous cytoplasmic proteins and precursors is discussed. The new challenges now are the development of food grade systems and the identification and optimization of host factors affecting heterologous protein production not only in L. lactis, but also in other LAB species.

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Section: Protein Conformation Rather Than Protein Size Can Impair Thesupporting

confidence: 72%

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et al. 2005

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“…The region N6 is followed by a Gly residue invariant in family GH68 (Fig. 3, line 1), which is essential for secretion efficiency and folding process of B. subtilis levansucrase 45. The regions N5 and N6 comprise the third conserved cluster, which we earlier described in sequences of families GH43 and GH68 20…”

Section: Discussionmentioning

confidence: 83%

?-Fructosidase superfamily: Homology with some ?-L-arabinases and ?-D-xylosidases

Naumoff

2000

Proteins

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“…The secretion of B. subtilis levansucrase is stimulated by growth of the cells in medium containing high concentrations of Fe 3ϩ , and in vitro refolding of B. subtilis levansucrase was greatly enhanced by this cation, even though Fe 3ϩ is not bound to the mature protein (49). Furthermore, several extracellular proteins from B. subtilis, such as levansucrase, neutral protease, and ␣-amylase, are calcium-binding proteins, and they require Ca 2ϩ for stability (212,275,313). Ca 2ϩ is trapped in the B. subtilis cell wall, thereby creating a microenvironment that must play an important role in the late steps of secretion (211).…”

Section: Extracytoplasmic Folding Catalystsmentioning

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

747

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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Bacillus subtilis levansucrase: amino acid substitutions at one site affect secretion efficiency and refolding kinetics mediated by metals

Cited by 29 publications

References 35 publications

Untitled

Untitled

?-Fructosidase superfamily: Homology with some ?-L-arabinases and ?-D-xylosidases

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

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