Loss of a functional mitochondrial pyruvate carrier in Komagataella phaffii does not improve lactic acid production from glycerol in aerobic cultivation

• Autor(a) principal: Junqueira, Ana Caroline de Oliveira
• Data de Publicação: 2023
• Outros Autores: Melo, Nadielle Tamires Moreira, Parachin, Nádia Skorupa, Paes, Hugo Costa
• Tipo de documento: Artigo
• Idioma: eng
• Título da fonte: Repositório Institucional da UnB
• Texto Completo: http://repositorio2.unb.br/jspui/handle/10482/46641; https://doi.org/10.3390/microorganisms11020483; https://orcid.org/0000-0003-0327-232X; https://orcid.org/0000-0002-1448-6852
• topic: Transportador de piruvato mitocondrial; Hemoglobina bacteriana; Glicerol

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.