Overexpression of the gene encoding alternative oxidase for enhanced glucose consumption in oxalic acid producing Aspergillus niger expressing oxaloacetate hydrolase gene

Isato Yoshioka, Keiichi Kobayashi, Kohtaro Kirimura

Research output: Contribution to journalArticle

Abstract

The filamentous fungus Aspergillus niger is a well-known hyper-producer of organic acids such as citric acid and oxalic acid. This fungus possesses the cyanide (CN)-insensitive respiration pathway consisting of alternative oxidase (EC 1.10.3.11; AOX), in addition to the cytochrome pathway. Since this CN-insensitive respiration pathway reoxidizes NADH without ATP production, it contributes to continuous glycolysis in A. niger. In this study, to show the availability of aoxA gene encoding AOX as a tool for metabolic engineering, we generated efficient oxalic acid (OA)-producers by genetic engineering of A. niger using aoxA gene. The OA-producing strain EOAH-1, generated by overexpression of the oxaloacetate hydrolase (EC 3.7.1.1; OAH) gene oahA in A. niger WU-2223L, produced 28 g/L OA from 30 g/L glucose during the 9-day cultivation period. Moreover, the strain EAOXOAH-1, generated by overexpression of both aoxA and oahA genes in strain WU-2223L, produced 28 g/L OA during the 7-day cultivation period. Strain EAOXOAH-1 showed higher glucose consumption rate than EOAH-1 did, indicating that overexpression of aoxA contributed to the acceleration of glucose consumption, and that the OA production period was shortened by 2 days. Thus, we clearly show that AOX gene must be an effective tool in metabolic engineering for efficient organic acids production from carbohydrates.

Original languageEnglish
JournalJournal of Bioscience and Bioengineering
DOIs
Publication statusAccepted/In press - 2019 Jan 1

Fingerprint

oxaloacetase
Oxalic Acid
Hydrolases
Gene encoding
Oxalic acid
Aspergillus niger
Aspergillus
Glucose
Genes
Metabolic engineering
Metabolic Engineering
Organic acids
Cyanides
Fungi
Respiration
Genetic engineering
Acids
Genetic Engineering
Adenosinetriphosphate
Citric acid

Keywords

  • Alternative oxidase
  • Aspergillus niger
  • Glycolysis
  • Metabolic engineering
  • Oxalic acid

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

Cite this

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title = "Overexpression of the gene encoding alternative oxidase for enhanced glucose consumption in oxalic acid producing Aspergillus niger expressing oxaloacetate hydrolase gene",
abstract = "The filamentous fungus Aspergillus niger is a well-known hyper-producer of organic acids such as citric acid and oxalic acid. This fungus possesses the cyanide (CN)-insensitive respiration pathway consisting of alternative oxidase (EC 1.10.3.11; AOX), in addition to the cytochrome pathway. Since this CN-insensitive respiration pathway reoxidizes NADH without ATP production, it contributes to continuous glycolysis in A. niger. In this study, to show the availability of aoxA gene encoding AOX as a tool for metabolic engineering, we generated efficient oxalic acid (OA)-producers by genetic engineering of A. niger using aoxA gene. The OA-producing strain EOAH-1, generated by overexpression of the oxaloacetate hydrolase (EC 3.7.1.1; OAH) gene oahA in A. niger WU-2223L, produced 28 g/L OA from 30 g/L glucose during the 9-day cultivation period. Moreover, the strain EAOXOAH-1, generated by overexpression of both aoxA and oahA genes in strain WU-2223L, produced 28 g/L OA during the 7-day cultivation period. Strain EAOXOAH-1 showed higher glucose consumption rate than EOAH-1 did, indicating that overexpression of aoxA contributed to the acceleration of glucose consumption, and that the OA production period was shortened by 2 days. Thus, we clearly show that AOX gene must be an effective tool in metabolic engineering for efficient organic acids production from carbohydrates.",
keywords = "Alternative oxidase, Aspergillus niger, Glycolysis, Metabolic engineering, Oxalic acid",
author = "Isato Yoshioka and Keiichi Kobayashi and Kohtaro Kirimura",
year = "2019",
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doi = "10.1016/j.jbiosc.2019.08.014",
language = "English",
journal = "Journal of Bioscience and Bioengineering",
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T1 - Overexpression of the gene encoding alternative oxidase for enhanced glucose consumption in oxalic acid producing Aspergillus niger expressing oxaloacetate hydrolase gene

AU - Yoshioka, Isato

AU - Kobayashi, Keiichi

AU - Kirimura, Kohtaro

PY - 2019/1/1

Y1 - 2019/1/1

N2 - The filamentous fungus Aspergillus niger is a well-known hyper-producer of organic acids such as citric acid and oxalic acid. This fungus possesses the cyanide (CN)-insensitive respiration pathway consisting of alternative oxidase (EC 1.10.3.11; AOX), in addition to the cytochrome pathway. Since this CN-insensitive respiration pathway reoxidizes NADH without ATP production, it contributes to continuous glycolysis in A. niger. In this study, to show the availability of aoxA gene encoding AOX as a tool for metabolic engineering, we generated efficient oxalic acid (OA)-producers by genetic engineering of A. niger using aoxA gene. The OA-producing strain EOAH-1, generated by overexpression of the oxaloacetate hydrolase (EC 3.7.1.1; OAH) gene oahA in A. niger WU-2223L, produced 28 g/L OA from 30 g/L glucose during the 9-day cultivation period. Moreover, the strain EAOXOAH-1, generated by overexpression of both aoxA and oahA genes in strain WU-2223L, produced 28 g/L OA during the 7-day cultivation period. Strain EAOXOAH-1 showed higher glucose consumption rate than EOAH-1 did, indicating that overexpression of aoxA contributed to the acceleration of glucose consumption, and that the OA production period was shortened by 2 days. Thus, we clearly show that AOX gene must be an effective tool in metabolic engineering for efficient organic acids production from carbohydrates.

AB - The filamentous fungus Aspergillus niger is a well-known hyper-producer of organic acids such as citric acid and oxalic acid. This fungus possesses the cyanide (CN)-insensitive respiration pathway consisting of alternative oxidase (EC 1.10.3.11; AOX), in addition to the cytochrome pathway. Since this CN-insensitive respiration pathway reoxidizes NADH without ATP production, it contributes to continuous glycolysis in A. niger. In this study, to show the availability of aoxA gene encoding AOX as a tool for metabolic engineering, we generated efficient oxalic acid (OA)-producers by genetic engineering of A. niger using aoxA gene. The OA-producing strain EOAH-1, generated by overexpression of the oxaloacetate hydrolase (EC 3.7.1.1; OAH) gene oahA in A. niger WU-2223L, produced 28 g/L OA from 30 g/L glucose during the 9-day cultivation period. Moreover, the strain EAOXOAH-1, generated by overexpression of both aoxA and oahA genes in strain WU-2223L, produced 28 g/L OA during the 7-day cultivation period. Strain EAOXOAH-1 showed higher glucose consumption rate than EOAH-1 did, indicating that overexpression of aoxA contributed to the acceleration of glucose consumption, and that the OA production period was shortened by 2 days. Thus, we clearly show that AOX gene must be an effective tool in metabolic engineering for efficient organic acids production from carbohydrates.

KW - Alternative oxidase

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