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Título: Loss of a functional mitochondrial pyruvate carrier in Komagataella phaffii does not improve lactic acid production from glycerol in aerobic cultivation
Autor(es): Junqueira, Ana Caroline de Oliveira
Melo, Nadielle Tamires Moreira
Parachin, Nádia Skorupa
Paes, Hugo Costa
ORCID: https://orcid.org/0000-0003-0327-232X
https://orcid.org/0000-0002-1448-6852
Afiliação do autor: University of Brasília, Department of Molecular Biology
Catholic University of Brasília, Genomic Sciences and Biotechnology
Gingko Bioworks
University of Brasília, University of Brasília Medical School, Clinical Medicine Division
Assunto: Transportador de piruvato mitocondrial
Hemoglobina bacteriana
Glicerol
Data de publicação: 15-Fev-2023
Editora: MDPI
Referência: JUNQUEIRA, Ana Caroline de Oliveira et al. Loss of a functional mitochondrial pyruvate carrier in Komagataella phaffii does not improve lactic acid production from glycerol in aerobic cultivation. Microorganisms, [S.l.], v. 11, n. 2, 483, 2023. DOI: https://doi.org/10.3390/microorganisms11020483. Disponível em: https://www.mdpi.com/2076-2607/11/2/483. Acesso em: 09 out. 2023.
Abstract: Cytosolic pyruvate is an essential metabolite in lactic acid production during microbial fermentation. However, under aerobiosis, pyruvate is transported to the mitochondrial matrix by the mitochondrial pyruvate carrier (MPC) and oxidized in cell respiration. Previous reports using Saccha- romyces cerevisiae or Aspergillus oryzae have shown that the production of pyruvate-derived chemicals is improved by deleting the MPC1 gene. A previous lactate-producing K. phaffii strain engineered by our group was used as a host for the deletion of the MPC1 gene. In addition, the expression of a bacterial hemoglobin gene under the alcohol dehydrogenase 2 promoter from Scheffersomyces stipitis, known to work as a hypoxia sensor, was used to evaluate whether aeration would supply enough oxygen to meet the metabolic needs during lactic acid production. However, unlike S. cerevisiae and A. oryzae, the deletion of Mpc1 had no significant impact on lactic acid production but negatively affected cell growth in K. phaffii strains. Furthermore, the relative quantification of the VHb gene revealed that the expression of hemoglobin was detected even in aerobic cultivation, which indicates that the demand for oxygen in the bioreactor could result in functional hypoxia. Overall, the results add to our previously published ones and show that blocking cell respiration using hypoxia is more suitable than deleting Mpc for producing lactic acid in K. phaffii.
Unidade Acadêmica: Instituto de Ciências Biológicas (IB)
Departamento de Biologia Celular (IB CEL)
Faculdade de Medicina (FMD)
Programa de pós-graduação: Programa de Pós-Graduação em Biologia Molecular
Licença: Copyright: © 2023 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https:// creativecommons.org/licenses/by/4.0).
DOI: https://doi.org/10.3390/microorganisms11020483
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