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

Autor: Reilly MC; Joint BioEnergy Institute, Emeryville, CA, 94608, USA.; Chemical and Biological Processes Development Group, Pacific Northwest National Laboratory, Richland, WA, 99352, USA., Kim J; Joint BioEnergy Institute, Emeryville, CA, 94608, USA.; Chemical and Biological Processes Development Group, Pacific Northwest National Laboratory, Richland, WA, 99352, USA., Lynn J; Joint BioEnergy Institute, Emeryville, CA, 94608, USA.; Naval Medical Research Unit Dayton, Wright-Patterson Air Force Base, Dayton, OH, 45433, USA., Simmons BA; Joint BioEnergy Institute, Emeryville, CA, 94608, USA.; Biological Systems and Engineering Division, Lawrence Berkeley National Laboratory, Berkeley, CA, 94720, USA., Gladden JM; Joint BioEnergy Institute, Emeryville, CA, 94608, USA.; Biomass Science and Conversion Technologies Department, Sandia National Laboratories, Livermore, CA, 94551, USA., Magnuson JK; Joint BioEnergy Institute, Emeryville, CA, 94608, USA.; Chemical and Biological Processes Development Group, Pacific Northwest National Laboratory, Richland, WA, 99352, USA., Baker SE; Joint BioEnergy Institute, Emeryville, CA, 94608, USA. scott.baker@pnnl.gov.; Biosystems Design and Simulation Group, Environmental Molecular Sciences Division, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. scott.baker@pnnl.gov.
Jazyk: angličtina
Zdroj: Applied microbiology and biotechnology [Appl Microbiol Biotechnol] 2018 Feb; Vol. 102 (4), pp. 1797-1807. Date of Electronic Publication: 2018 Jan 06.
DOI: 10.1007/s00253-017-8717-3
Abstrakt: 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.
Databáze: MEDLINE
Externí odkaz:
Nepřihlášeným uživatelům se plný text nezobrazuje