Functional analysis of the mitochondrial alternative oxidase gene (aox1) from Aspergillus niger CGMCC 10142 and its effects on citric acid production

Applied Microbiology and Biotechnology
Li HouDepei Wang

Abstract

In this work, we constructed the aox1 disruption strains 3-4 and 4-10, as well as the aox1 overexpression strains 72 and 102 in Aspergillus niger. The energy metabolism, EMP, TCA pathways, and flux were investigated for the citric acid (CA) overproduction via the aox1 overexpression among them. As expected, the overexpression of the aox1 gene enabled a higher growth rate than that of the rate of its parent strain in medium with respiratory chain inhibitors. In liquefied corn starch medium supplemented with 0.2 μg/mL antimycin A, the CA production of the overexpression strain 102 reached up to 169.1 g/L, whereas the highest value of the parent strain was 158.9 g/L. For the perspective of the aox1 disruption strain 4-10, the yield of CA dropped to 125.6 g/L, and the loose mycelial pellets forming in the medium also revealed that the fundamentally important role of AOX in A. niger lies in the resistance to oxidative stress under fully aerobic conditions. Based on real-time qPCR gene expression analysis and measurement of intracellular ATP and NADH levels, we came to a conclusion that the higher NADH oxidation rate resulting from the overexpression of the aox1 gene mainly contributed to rate-limited step's acceleration and strength...Continue Reading

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Citations

Feb 14, 2019·Applied Microbiology and Biotechnology·Levente Karaffa, Christian P Kubicek
Nov 14, 2018·Applied Microbiology and Biotechnology·Wei HuJi-Hong Chen
Oct 13, 2020·Critical Reviews in Microbiology·Bikash Chandra Behera
Jan 15, 2021·Scientific Reports·Jorgelindo da Veiga MoreiraSabine Peres

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Methods Mentioned

BETA
genetic modifications
Assay
Enzyme-Linked Immunosorbent Assay
PCR
spore
reverse transcription PCR
genetic modification

Software Mentioned

Clone Manager
Origin
Excel

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