Siedler, Solvej; Bringer, Stephanie; Polen, Tino; Bott, Michael published an article in 2014, the title of the article was NADPH-dependent reductive biotransformation with Escherichia coli and its pfkA deletion mutant: Influence on global gene expression and role of oxygen supply.Application of 3976-69-0 And the article contains the following content:
An Escherichia coli ΔpfkA mutant lacking the major phosphofructokinase possesses a partially cyclized pentose phosphate pathway leading to an increased NADPH per glucose ratio. This effect decreases the amount of glucose required for NADPH regeneration in reductive biotransformations, such as the conversion of Me acetoacetate (MAA) to (R)-Me 3-hydroxybutyrate (MHB) by an alc. dehydrogenase from Lactobacillus brevis. Here, global transcriptional analyses were performed to study regulatory responses during reductive biotransformation. DNA microarray anal. revealed amongst other things increased expression of soxS, supporting previous results indicating that a high NADPH demand contributes to the activation of SoxR, the transcriptional activator of soxS. Furthermore, several target genes of the ArcAB two-component system showed a lower mRNA level in the reference strain than in the ΔpfkA mutant, pointing to an increased QH2/Q ratio in the reference strain. This prompted us to analyze yields and productivities of MAA reduction to MHB under different oxygen regimes in a bioreactor. Under anaerobic conditions, the specific MHB production rates of both strains were comparable (7.4 ± 0.2 mmolMHB h-1 gcdw-1) and lower than under conditions of 15% dissolved oxygen, where those of the reference strain (12.8 mmol h-1 gcdw-1) and of the ΔpfkA mutant (11.0 mmol h-1 gcdw-1) were 73% and 49% higher. While the oxygen transfer rate (OTR) of the reference strain increased after the addition of MAA, presumably due to the oxidation of the acetate accumulated before MAA addition, the OTR of the ΔpfkA strain strongly decreased, indicating a very low respiration rate despite sufficient oxygen supply. The latter effect can likely be attributed to a restricted conversion of NADPH into NADH via the soluble transhydrogenase SthA, as the enzyme is outcompeted in the presence of MAA by the recombinant NADPH-dependent alc. dehydrogenase. The differences in respiration rates can explain the suggested higher ArcAB activity in the reference strain. Biotechnol. Bioeng. 2014;9999: 1-9. © 2014 Wiley Periodicals, Inc. The experimental process involved the reaction of (R)-Methyl 3-hydroxybutanoate(cas: 3976-69-0).Application of 3976-69-0
The Article related to escherichia reductive biotransformation oxygen supply gene expression, dna microarrays, escherichia coli, stha transhydrogenase, phosphofructokinase, reductive whole-cell biotransformation, resting cells and other aspects.Application of 3976-69-0
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