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产朊假丝酵母重组菌株中过表达异源酶的蛋白质谱。

Protein profile in Aspergillus nidulans recombinant strains overproducing heterologous enzymes.

机构信息

Department of Biochemistry and Tissue Biology, Institute of Biology, University of Campinas (UNICAMP), Campinas, SP, Brazil.

Department of Internal Medicine, Faculty of Medical Sciences, University of Campinas (UNICAMP), Campinas, SP, Brazil.

出版信息

Microb Biotechnol. 2018 Mar;11(2):346-358. doi: 10.1111/1751-7915.13027. Epub 2018 Jan 8.

Abstract

Filamentous fungi are robust cell factories and have been used for the production of large quantities of industrially relevant enzymes. However, the production levels of heterologous proteins still need to be improved. Therefore, this article aimed to investigate the global proteome profiling of Aspergillus nidulans recombinant strains in order to understand the bottlenecks of heterologous enzymes production. About 250, 441 and 424 intracellular proteins were identified in the control strain Anid_pEXPYR and in the recombinant strains Anid_AbfA and Anid_Cbhl respectively. In this context, the most enriched processes in recombinant strains were energy pathway, amino acid metabolism, ribosome biogenesis, translation, endoplasmic reticulum and oxidative stress, and repression under secretion stress (RESS). The global protein profile of the recombinant strains Anid_AbfA and Anid_Cbhl was similar, although the latter strain secreted more recombinant enzyme than the former. These findings provide insights into the bottlenecks involved in the secretion of recombinant proteins in A. nidulans, as well as in regard to the rational manipulation of target genes for engineering fungal strains as microbial cell factories.

摘要

丝状真菌是强大的细胞工厂,已被用于大量生产工业相关的酶。然而,异源蛋白的生产水平仍有待提高。因此,本文旨在研究构巢曲霉重组菌株的全蛋白质组谱,以了解异源酶生产的瓶颈。在对照菌株 Anid_pEXPYR 和重组菌株 Anid_AbfA 和 Anid_Cbhl 中分别鉴定了约 250、441 和 424 种细胞内蛋白质。在这种情况下,重组菌株中最丰富的过程是能量途径、氨基酸代谢、核糖体生物发生、翻译、内质网和氧化应激,以及在分泌应激下的抑制(RESS)。重组菌株 Anid_AbfA 和 Anid_Cbhl 的全蛋白质谱相似,尽管后者菌株分泌的重组酶比前者多。这些发现为理解构巢曲霉中重组蛋白分泌的瓶颈提供了线索,也为理性操纵目标基因以工程化真菌菌株作为微生物细胞工厂提供了线索。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/17e1/5812239/e31d2cc3e49d/MBT2-11-346-g001.jpg

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