Glucoamylase gene fusions alleviate limitations for protein production in Aspergillus awamori at the transcriptional and (post) translational levels

@article{Gouka1997GlucoamylaseGF,
  title={Glucoamylase gene fusions alleviate limitations for protein production in Aspergillus awamori at the transcriptional and (post) translational levels},
  author={Robin Gouka and Peter J. Punt and Cees A.M.J.J. van den Hondel},
  journal={Applied and Environmental Microbiology},
  year={1997},
  volume={63},
  pages={488 - 497}
}
In this study we have analyzed the effects of a glucoamylase gene fusion on the mRNA levels and protein levels for the human interleukin-6 gene (hil6) and the guar alpha-galactosidase gene (aglA). Previously it was shown that production of nonfused alpha-galactosidase and hIL-6 in Aspergillus awamori was limited at transcriptional and (post)translational levels, respectively (R. J. Gouka, P. J. Punt, J. G. M. Hessing, and C. A. M. J. J. van den Hondel, Appl. Environ. Microbiol. 62:1951-1957… 
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References

SHOWING 1-10 OF 45 REFERENCES
Analysis of heterologous protein production in defined recombinant Aspergillus awamori strains
TLDR
The data suggest that intracellular protein degradation is the most likely explanation for the low levels of secreted human interleukin 6.
An expression system based on the promoter region of the Aspergillus awamori 1,4-β-endoxylanase A gene
TLDR
A new, highly inducible fungal promoter derived from the Aspergillus awamori 1,4-β-endoxylanase A (exlA) gene is described and induction analyses demonstrated that induction was dependent on the presence of D-xylose in the medium.
Secretion of the alpha-galactosidase from Cyamopsis tetragonoloba (guar) by Bacillus subtilis
TLDR
The activity of this Bacillus sp.-produced enzyme was similar to that of the glycosylated enzyme purified from guar seeds, indicating that glycosolation has no essential function for enzyme activity.
Expression of the α‐galactosidase from Cyamopsis tetragonoloba (guar) by Hansenula polymorpha
TLDR
The methylotrophic yeast Hansenula polymorpha, a host organism for the production of heterologous proteins, has been applied to produce the α‐galactosidase from the plant Cyamopsis tetragonoloba (guar) to show that the S. cerevisiae invertase signal sequence was correctly processed and restored the specific activity completely.
Efficient KEX2-Like Processing of a Glucoamylase-Interleukin-6 Fusion Protein by Aspergillus Nidulans and Secretion of Mature Interleukin-6
We have designed an expression vector for the secretion of human interleukin-6 (hIL-6) in which the mature protein is fused through a spacer peptide, contain-ing a KEX-2 like protein processing
Aspergillus Niger var. Awamori as a Host for the Expression of Heterologous Genes
TLDR
Many of the molecular details and principles which have been elucidated in yeast and in mammalian cell systems appear to be applicable to the study of heterologous gene expression and protein secretion in filamentous fungi as well.
Expression of tetanus toxin fragment C in yeast: gene synthesis is required to eliminate fortuitous polyadenylation sites in AT-rich DNA.
TLDR
Intracellular fragment C was as effective in vaccinating mice against tetanus authentic fragment C, though the glycosylated material was inactive, though it was rendered fully active by de-glycosylation.
Transcriptional and post-transcriptional events affect the production of secreted hen egg white lysozyme by Aspergillus niger.
TLDR
Results suggest that post-transcriptional events significantly affect final secreted protein yields in Aspergillus niger transformants containing either the hen egg-white lysozyme cDNA gene or a glucoamylase-HEWL gene fusion.
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