Inhibition of alternative respiration system of Scheffersomyces stipitis and effect on glucose or xylose fermentation

Scheffersomyces stipitis is a Crabtree-negative pentose fermenting yeast, which shows a complex respiratory system involving a cytochrome and an alternative salicylhydroxamic acid (SHAM)-sensitive respiration mechanism that is poorly understood. This work aimed to investigate the role of the antimycin A (AA) sensitive respiration and SHAM-sensitive respiration in the metabolism of xylose and glucose by S. stipitis, upon different agitation conditions. Inhibition of the SHAM-sensitive respiration caused a significant (p < 0.05) decrease in glycolytic flux and oxygen consumption when using glucose and xylose under agitation conditions, but without agitation, only a mild reduction was observed. The combination of SHAM and AA abolished respiration, depleting the glycolytic flux using both carbon sources tested, leading to increased ethanol production of 21.05 g/L at 250 rpm for 0.5 M glucose, and 8.3 g/L ethanol using xylose. In contrast, inhibition of only the AA-sensitive respiration, caused increased ethanol production to 30 g/L using 0.5 M glucose at 250 rpm, and 11.3 g/L from 0.5 M xylose without agitation. Results showed that ethanol production can be induced by respiration inhibition, but the active role of SHAM-sensitive respiration should be considered to investigate better conditions to increase and optimize yields.

Activation of Alternative Respiration with Internal Electron Acceptor upon Anaerobic Glucose Utilization in Escherichia coli strains with Impaired Fermentation Ability

The activation of alternative respiration with an internal electron acceptor upon anaerobic utilization of glucose in Escherichia coli strains with the impaired fermentation ability has been studied. It was found out that the respiration processes utilizing pyruvic acid as an endogenous electron acceptor can markedly contribute to the maintenance of the anaerobic redox balance in E. coli strains deficient in mixed acid fermentation pathways. The sequential inactivation of the pathways of anaerobic dissimilation of pyruvate and impairment of the functionality of the reductive branch of the tricarboxylic acid cycle led to the increase in the contribution of the respiratory formation of lactic acid and alanine to the biosynthesis of the reduced products of anaerobic glucose utilization by the strains from 11 to 54%. The analysis of the enantiomeric composition of the lactic acid and alanine secreted by the strains demonstrated that D-lactate dehydrogenase (Dld), L-lactate dehydrogenase (LctD), and D-alanine dehydrogenase (DadA) participated in the biosynthesis of the respective compounds. Escherichia coli, glucose, fermentation, respiration, metabolic engineering, pyruvate, lactate, alanine. The work was supported by a grant from the Russian Foundation for Basic Research (Project no. 18-04-01222)

Differential root transcriptomics in a polyploid non-model crop: the importance of respiration during osmotic stress

To explore the transcriptomic global response to osmotic stress in roots, 18 mRNA-seq libraries were generated from three triploid banana genotypes grown under mild osmotic stress (5% PEG) and control conditions. Illumina sequencing produced 568 million high quality reads, of which 70–84% were mapped to the banana diploid reference genome. Using different uni- and multivariate statistics, 92 genes were commonly identified as differentially expressed in the three genotypes. Using our in house workflow to analyze GO enriched and underlying biochemical pathways, we present the general processes affected by mild osmotic stress in the root and focus subsequently on the most significantly overrepresented classes associated with: respiration, glycolysis and fermentation. We hypothesize that in fast growing and oxygen demanding tissues, mild osmotic stress leads to a lower energy level, which induces a metabolic shift towards (i) a higher oxidative respiration, (ii) alternative respiration and (iii) fermentation. To confirm the mRNA-seq results, a subset of twenty up-regulated transcripts were further analysed by RT-qPCR in an independent experiment at three different time points. The identification and annotation of this set of genes provides a valuable resource to understand the importance of energy sensing during mild osmotic stress.