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Title: Forward genetics screen coupled with whole-genome resequencing identifies novel gene targets for improving heterologous enzyme production in Aspergillus niger

Plant biomass, once reduced to its composite sugars, can be converted to fuel substitutes. One means of overcoming the recalcitrance of lignocellulose is pretreatment followed by enzymatic hydrolysis. However, currently available commercial enzyme cocktails are inhibited in the presence of residual pretreatment chemicals. Recent studies have identified a number of cellulolytic enzymes from bacteria that are tolerant to pretreatment chemicals such as ionic liquids. The challenge now is generation of these enzymes in copious amounts, an arena where fungal organisms such as Aspergillus niger have proven efficient. Fungal host strains still need to be engineered to increase production titers of heterologous protein over native enzymes, which has been a difficult task. Here, we developed a forward genetics screen coupled with whole-genome resequencing to identify specific lesions responsible for a protein hyper-production phenotype in A. niger. This strategy successfully identified novel targets, including a low-affinity glucose transporter, MstC, whose deletion significantly improved secretion of recombinant proteins driven by a glucoamylase promoter.
Authors:
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Publication Date:
Report Number(s):
PNNL-SA-132880
Journal ID: ISSN 0175-7598; 830403000
Grant/Contract Number:
AC02-05CH11231; AC05-76RL01830
Type:
Published Article
Journal Name:
Applied Microbiology and Biotechnology
Additional Journal Information:
Journal Volume: 102; Journal Issue: 4; Journal ID: ISSN 0175-7598
Publisher:
Springer
Research Org:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Sponsoring Org:
USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Aspergillus niger; Heterologous protein expression; Forward mutagenesis; Whole-genome resequencing; MstC
OSTI Identifier:
1416031
Alternate Identifier(s):
OSTI ID: 1431407; OSTI ID: 1433118