Identification of a specific exporter that enables high production of aconitic acid in Aspergillus pseudoterreus.

Autor: Deng S; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: shuang.deng@pnnl.gov., Kim J; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: joonhoon.kim@pnnl.gov., Pomraning KR; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: kyle.pomraning@pnnl.gov., Gao Y; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: Yuqian.gao@pnnl.gov., Evans JE; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: James.Evans@pnnl.gov., Hofstad BA; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: beth.hofstad@pnnl.gov., Dai Z; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: ziyu.dai@pnnl.gov., Webb-Robertson BJ; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: bobbie-jo.webb-robertson@pnnl.gov., Powell SM; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: samantha.powell@pnnl.gov., Novikova IV; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: irina.novikova@pnnl.gov., Munoz N; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: nathalie.munozmunoz@pnnl.gov., Kim YM; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: young-mo.kim@pnnl.gov., Swita M; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: marie.swita@pnnl.gov., Robles AL; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: ana.robles@pnnl.gov., Lemmon T; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: Teresa.lemmon@pnnl.gov., Duong RD; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: rylan.duong@pnnl.gov., Nicora C; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: carrie.nicora@pnnl.gov., Burnum-Johnson KE; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Earth and Biological Sciences Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA. Electronic address: Kristin.burnum-johnson@pnnl.gov., Magnuson J; DOE Agile Biofoundry, Emeryville, CA, 94608, USA; Energy and Environment Directorate, Pacific Northwest National Laboratory, Richland, WA, 99352, USA.
Jazyk: angličtina
Zdroj: Metabolic engineering [Metab Eng] 2023 Nov; Vol. 80, pp. 163-172. Date of Electronic Publication: 2023 Sep 29.
DOI: 10.1016/j.ymben.2023.09.011
Abstrakt: Aconitic acid is an unsaturated tricarboxylic acid that is attractive for its potential use in manufacturing biodegradable and biocompatible polymers, plasticizers, and surfactants. Previously Aspergillus pseudoterreus was engineered as a platform to produce aconitic acid by deleting the cadA (cis-aconitic acid decarboxylase) gene in the itaconic acid biosynthetic pathway. In this study, the aconitic acid transporter gene (aexA) was identified using comparative global discovery proteomics analysis between the wild-type and cadA deletion strains. The protein AexA belongs to the Major Facilitator Superfamily (MFS). Deletion of aexA almost abolished aconitic acid secretion, while its overexpression led to a significant increase in aconitic acid production. Transportation of aconitic acid across the plasma membrane is a key limiting step in its production. In vitro, proteoliposome transport assay further validated AexA's function and substrate specificity. This research provides new approaches to efficiently pinpoint and characterize exporters of fungal organic acids and accelerate metabolic engineering to improve secretion capability and lower the cost of bioproduction.
(Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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