Enhanced production and secretion of heterologous proteins by the filamentous fungus Aspergillus oryzae via disruption of vacuolar protein sorting receptor gene Aovps10.

Applied and Environmental Microbiology
Jaewoo YoonKatsuhiko Kitamoto

Abstract

Filamentous fungi have received attention as hosts for heterologous protein production because of their high secretion capability and eukaryotic posttranslational modifications. However, despite these positive attributes, a bottleneck in posttranscriptional processing limits protein yields. The vacuolar protein sorting gene VPS10 encodes a sorting receptor for the recognition and delivery of several yeast vacuolar proteins. Although it can also target recombinant and aberrant proteins for vacuolar degradation, there is limited knowledge of the effect of its disruption on heterologous protein production. In this study, cDNA encoding AoVps10 from the filamentous fungus Aspergillus oryzae was cloned and sequenced. Microscopic observation of the transformant expressing AoVps10 fused with enhanced green fluorescent protein showed that the fusion protein localized at the Golgi and prevacuolar compartments. Moreover, disruption of the Aovps10 gene resulted in missorting and secretion of vacuolar carboxypeptidase AoCpyA into the medium, indicating that AoVps10 is required for sorting of vacuolar proteins to vacuoles. To investigate the extracellular production levels of heterologous proteins, DeltaAovps10 mutants expressing either bovi...Continue Reading

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Citations

Aug 9, 2011·Applied Microbiology and Biotechnology·Ayako OhnoKatsuhiko Kitamoto
Feb 22, 2012·Applied Microbiology and Biotechnology·Martin PfefferDiethard Mattanovich
Oct 14, 2011·Yi chuan = Hereditas·Xian-Zhong ChenZheng-Xiang Wang
Jan 19, 2016·Journal of Bioscience and Bioengineering·Hiroshi KikukawaJun Ogawa
Oct 31, 2012·Molecular Microbiology·Robert-Jan BleichrodtHan A B Wösten
Oct 16, 2015·Journal of Bioscience and Bioengineering·Hidetoshi NakamuraKatsuhiko Kitamoto
Dec 20, 2014·BMC Genomics·Tatiana Q AguiarLucília Domingues
Jul 23, 2011·Journal of Biotechnology·Jozef AnnéKristel Bernaerts
May 24, 2016·The Journal of General and Applied Microbiology·Chihiro KadookaTaiki Futagami
Jun 10, 2020·Fungal biology and biotechnology·Hung Hiep HuynhJun-Ichi Maruyama
Jan 14, 2011·Traffic·Areti Pantazopoulou, Miguel A Peñalva
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Apr 24, 2020·Frontiers in Bioengineering and Biotechnology·Qin WangHan Xiao
Jan 19, 2020·Protein Expression and Purification·Uriel Rojas-SánchezMiguel A Gómez-Lim

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