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

Reilly, Morgann C.; Kim, Joonhoon; Lynn, Jed; Simmons, Blake A.; Gladden, John M.; Magnuson, Jon K.; Baker, Scott E.

doi:10.1007/s00253-017-8717-3

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

Journal Article · Sat Jan 06 00:00:00 EST 2018 · Applied Microbiology and Biotechnology

DOI:https://doi.org/10.1007/s00253-017-8717-3· OSTI ID:1416031

Reilly, Morgann C. ^[1]; Kim, Joonhoon ^[1]; Lynn, Jed ^[2]; Simmons, Blake A. ^[3]; Gladden, John M. ^[4]; Magnuson, Jon K. ^[1];

^[1]

Joint BioEnergy Institute, Emeryville, CA (United States); Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
Joint BioEnergy Institute, Emeryville, CA (United States); Wright-Patterson Air Force Base, Dayton, OH (United States)
Joint BioEnergy Institute, Emeryville, CA (United States); Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States)
Joint BioEnergy Institute, Emeryville, CA (United States); Sandia National Lab. (SNL-CA), Livermore, CA (United States)

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.

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Research Organization:: Pacific Northwest National Laboratory (PNNL), Richland, WA (United States); Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)

Sponsoring Organization:: USDOE Office of Science (SC), Biological and Environmental Research (BER)

Grant/Contract Number:: AC02-05CH11231; AC05-76RL01830

OSTI ID:: 1416031

Alternate ID(s):: OSTI ID: 1420441; OSTI ID: 1431407; OSTI ID: 1433118

Report Number(s):: PNNL-SA-132880; PII: 8717

Journal Information:: Applied Microbiology and Biotechnology, Journal Name: Applied Microbiology and Biotechnology Vol. 102 Journal Issue: 4; ISSN 0175-7598

Publisher:: SpringerCopyright Statement

Country of Publication:: Germany

Language:: English

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Cited by: 11 works

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59 BASIC BIOLOGICAL SCIENCES
Aspergillus niger
Heterologous protein expression
Forward mutagenesis
Whole-genome resequencing
MstC

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

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