Efficient secretion of bacillus subtilis levanase by saccharomyces cerevisiae

Wanker, Erich E.; Klingsbichel, E.; Schwab, Helmut

doi:10.1016/0378-1119(95)00284-d

Cited by 9 publications

(5 citation statements)

References 17 publications

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“…Signal sequences of gram-negative bacteria are recognized by Sec translocons of gram-positive bacteria and vice versa (22,25,32), and even eukaryotic signal peptides of the Sec-related pathway for protein import into the endoplasmatic reticulum and bacterial Sec signal peptides are interchangeable (12,40). The observed species specificity of pre-GFOR export via the Tat pathway is therefore in marked contrast to the general Sec pathway.…”

Section: Discussionmentioning

confidence: 98%

Specificity of Signal Peptide Recognition in Tat-Dependent Bacterial Protein Translocation

et al. 2001

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The bacterial twin arginine translocation (Tat) pathway translocates across the cytoplasmic membrane folded proteins which, in most cases, contain a tightly bound cofactor. Specific amino-terminal signal peptides that exhibit a conserved amino acid consensus motif, S/T-R-R-X-F-L-K, direct these proteins to the Tat translocon. The glucose-fructose oxidoreductase (GFOR) of Zymomonas mobilis is a periplasmic enzyme with tightly bound NADP as a cofactor. It is synthesized as a cytoplasmic precursor with an amino-terminal signal peptide that shows all of the characteristics of a typical twin arginine signal peptide. However, GFOR is not exported to the periplasm when expressed in the heterologous host Escherichia coli, and enzymatically active pre-GFOR is found in the cytoplasm. A precise replacement of the pre-GFOR signal peptide by an authentic E. coli Tat signal peptide, which is derived from pre-trimethylamine N-oxide (TMAO) reductase (TorA), allowed export of GFOR, together with its bound cofactor, to the E. coli periplasm. This export was inhibited by carbonyl cyanide m-chlorophenylhydrazone, but not by sodium azide, and was blocked in E. coli tatC and tatAE mutant strains, showing that membrane translocation of the TorA-GFOR fusion protein occurred via the Tat pathway and not via the Sec pathway. Furthermore, tight cofactor binding (and therefore correct folding) was found to be a prerequisite for proper translocation of the fusion protein. These results strongly suggest that Tat signal peptides are not universally recognized by different Tat translocases, implying that the signal peptides of Tatdependent precursor proteins are optimally adapted only to their cognate export apparatus. Such a situation is in marked contrast to the situation that is known to exist for Sec-dependent protein translocation.

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Section: Discussionmentioning

confidence: 98%

Specificity of Signal Peptide Recognition in Tat-Dependent Bacterial Protein Translocation

et al. 2001

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“…This glycosylated endoxylanase was predominantly found in the culture medium of S. cerevisiae SEY2102 or 2805 harboring pAEDX-1, suggesting that the signal peptide of Bacillus endoxylanase functioned well in the yeast secretory pathway. This high secretion efficiency was also observed in the expression of B. subtilis levanase gene [23] and B. macerans cyclodextrin glucanotransferase gene in S. cerevisiae [24].…”

Section: Analysis Of Extracellular Glycosylated Endoxylanasementioning

confidence: 84%

Thermostability and xylan-hydrolyzing property of endoxylanase expressed in yeast Saccharomyces cerevisiae

Lee

Heo

Lee

et al. 2009

Biotechnol Bioproc E

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The endoxylanase gene (ñóå_, GeneBank access code U51675), including its signal sequence, from _~Åáääìë spp. was amplified and connected in frame downstream of yeast ^aeN promoter and then the resulting plasmid, pAEDX-1, was introduced into p~ÅÅÜ~êçãóÅÉë=ÅÉêÉîáëá~É. When the yeast transformants were grown on YPD medium, the majority of endoxylanase activity was detected in the extracellular culture medium, indicating that the signal peptide of _~Åáääìë endoxylanase functioned well in yeast. In the batch cultivation of yeast transformants, the total expression level of endoxylanase and secretion efficiency were measured to be about 9.8 U/mL and 66.2%, respectively. The extracellular endoxylanase expressed in yeast showed an enhanced thermal stability due to the k-linked glycosylation. Through the hydrolysis of birchwood xylan with the endoxylanase, it was found that xylobiose and xylotriose were produced as major products with equimolar ratio. © KSBB hÉóïçêÇëW=båÇçñóä~å~ëÉ, Saccharomyces cerevisiaeI íÜÉêãçëí~Äáäáíó, ñóäçÄáçëÉI=ñóäçíêáçëÉ=

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“…Modification of the hydrophobic core of the human apolipoprotein B signal sequence fused to invertase has been shown to affect the translocation efficiency of the protein in yeast (Sturley et al 1994). The signal sequence of the B. subtilis levanase which exhibits a high hydrophobic degree (Wanke et al 1995) was recently shown to be efficiently secreted by yeast in contrast to B. subtilis neutral protease (Wang and Devenish, 1993; Table 4). Moreover, efficient secretion of the yeast carboxypeptidase Y from mammalian cells also depends on the overall hydrophobicity of the h-domain in the signal sequence (Bird et al 1990).…”

Section: Discussionmentioning

confidence: 99%

“…Data on secretion in S. cerevisiae are from Romanos et al (1992) except for heterologous secretion of neutral protease (Wang and Devenish, 1993), of a-galactosidase (Harmsen et al, 1993) and of levanase (Wanke et a/., 1995). aMature part of the protein; bpropeptide part of the protein; "heterologous secretion under the control of the invertase signal sequence; dheterologous or homologous secretion of proteins under the control of their own secretion signal.…”

mentioning

confidence: 99%

The targeting of bacillus subtilis levansucrase in yeast is correlated to both the hydrophobicity of the signal peptide and the net charge of the N‐terminus mature part

Scotti

Præstegaard

Chambert

et al. 1996

Yeast

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Efficient secretion of bacillus subtilis levanase by saccharomyces cerevisiae

Cited by 9 publications

References 17 publications

Specificity of Signal Peptide Recognition in Tat-Dependent Bacterial Protein Translocation

Specificity of Signal Peptide Recognition in Tat-Dependent Bacterial Protein Translocation

Thermostability and xylan-hydrolyzing property of endoxylanase expressed in yeast Saccharomyces cerevisiae

The targeting of bacillus subtilis levansucrase in yeast is correlated to both the hydrophobicity of the signal peptide and the net charge of the N‐terminus mature part

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