de Oliveira Junqueira Ana Caroline, Moreira Melo Nadielle Tamires, Skorupa Parachin Nádia, Costa Paes Hugo
Department of Molecular Biology, University of Brasília, Brasília 70790-900, Brazil.
Genomic Sciences and Biotechnology, Catholic University of Brasília, Brasília 70790-160, Brazil.
Microorganisms. 2023 Feb 15;11(2):483. doi: 10.3390/microorganisms11020483.
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 or have shown that the production of pyruvate-derived chemicals is improved by deleting the gene. A previous lactate-producing strain engineered by our group was used as a host for the deletion of the gene. In addition, the expression of a bacterial hemoglobin gene under the alcohol dehydrogenase 2 promoter from , 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 and , the deletion of Mpc1 had no significant impact on lactic acid production but negatively affected cell growth in strains. Furthermore, the relative quantification of the 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 .
胞质丙酮酸是微生物发酵过程中乳酸生产的必需代谢物。然而,在有氧条件下,丙酮酸通过线粒体丙酮酸载体(MPC)转运到线粒体基质中,并在细胞呼吸中被氧化。以前使用或的报告表明,通过缺失基因可提高丙酮酸衍生化学品的产量。我们小组之前构建的一株产乳酸菌株被用作缺失该基因的宿主。此外,来自的已知作为缺氧传感器的乙醇脱氢酶2启动子下的细菌血红蛋白基因的表达,用于评估通气是否能提供足够的氧气以满足乳酸生产过程中的代谢需求。然而,与和不同,Mpc1的缺失对乳酸生产没有显著影响,但对菌株的细胞生长有负面影响。此外,基因的相对定量显示,即使在有氧培养中也检测到血红蛋白的表达,这表明生物反应器中的氧气需求可能导致功能性缺氧。总体而言,这些结果补充了我们之前发表的结果,并表明在中使用缺氧阻断细胞呼吸比缺失Mpc更适合生产乳酸。