Alone at Last! - Heterologous Expression of a Single Gene is Sufficient for Establishing the Five-step Weimberg Pathway in

Overview

Journal Metab Eng Commun

Specialty Biochemistry

Date 2019 Apr 25

PMID 31016135

Citations 6

Authors

Christian Brusseler

Anja Spath

Sascha Sokolowsky

Jan Marienhagen

Affiliations

Soon will be listed here.

Abstract

can grow on d-xylose as sole carbon and energy source via the five-step Weimberg pathway when the pentacistronic operon from is heterologously expressed. More recently, it could be demonstrated that the wild type accumulates the Weimberg pathway intermediate d-xylonate when cultivated in the presence of d-xylose. Reason for this is the activity of the endogenous dehydrogenase IolG, which can also oxidize d-xylose. This raised the question whether additional endogenous enzymes in contribute to the catabolization of d-xylose via the Weimberg pathway. In this study, analysis of the genome in combination with systematic reduction of the heterologous operon revealed that the hitherto unknown and endogenous dehydrogenase KsaD (Cg0535) can also oxidize α-ketoglutarate semialdehyde to the tricarboxylic acid cycle intermediate α-ketoglutarate, the final enzymatic step of the Weimberg pathway. Furthermore, heterologous expression of either or , encoding for the two dehydratases of the Weimberg pathway in , is sufficient for enabling to grow on d-xylose as sole carbon and energy source. Finally, several variants for the carbon-efficient microbial production of α-ketoglutarate from d-xylose were constructed. In comparison to cultivation solely on d-glucose, the best strain accumulated up to 1.5-fold more α-ketoglutarate in d-xylose/d-glucose mixtures.

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