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Characterization of Two Dehydrogenases from Gluconobacter oxydans Involved in the Transformation of Patulin to Ascladiol.

ABSTRACT: Patulin is a mycotoxin that primarily contaminate apples and apple products. Whole cell or cell-free extracts of Gluconobacter oxydans ATCC 621 were able to transform patulin to E-ascladiol. Proteins from cell-free extracts were separated by anion exchange chromatography and fractions with patulin transformation activity were subjected to peptide mass fingerprinting, enabling the identification of two NADPH dependent short chain dehydrogenases, GOX0525 and GOX1899, with the requisite activity. The genes encoding these enzymes were expressed in E. coli and purified. Kinetic parameters for patulin reduction, as well as pH profiles and thermostability were established to provide further insight on the potential application of these enzymes for patulin detoxification.

SUBMITTER: Chan ETS

PROVIDER: S-EPMC9323132 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Enhanced production of l-sorbose by systematic engineering of dehydrogenases in <i>Gluconobacter oxydans</i>.

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High-Throughput Screening of a 2-Keto-L-Gulonic Acid-Producing Gluconobacter oxydans Strain Based on Related Dehydrogenases.

Project description:High-throughput screening is a powerful tool for discovering strains in the natural environment that may be suitable for target production. Herein, a novel enzyme-based high-throughput screening method was developed for rapid screening of strains overproducing 2-keto-L-gulonic acid (2-KLG). The screening method detects changes in the fluorescence of reduced nicotinamide adenine dinucleotide (NADH) at 340 nm using a microplate reader when 2-KLG is degraded by 2-KLG reductase. In this research, three different 2-KLG reductases were expressed, purified, and studied. The 2-KLG reductase from Aspergillus niger were selected as the best appropriate reductase to establishment the method for its high activity below pH 7. Using the established method, and coupled with fluorescence-activated cell sorting, we achieved a high 2-KLG-producing strain of Gluconobacter oxydans WSH-004 from soil. When cultured with D-sorbitol as the substrate, the 2-KLG yield was 2.5 g/L from 50 g/L D-sorbitol without any side products. Compared with other reported screening methods, our enzyme-based method is more efficient and accurate for obtaining high-producing 2-KLG strains, and it is also convenient and cost-effective. The method is broadly applicable for screening keto acids and other products that can be oxidized via nicotinamide adenine dinucleotide (NAD+) or nicotinamide adenine dinucleotide phosphate (NADP+).

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A novel strain of acetic acid bacteria Gluconobacter oxydans FBFS97 involved in riboflavin production.

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Project description:Gluconobacter oxydans Raw sequence reads

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Project description:Industrially useful bacterium

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Global mRNA decay in Gluconobacter oxydans 621H

Project description:In this study we conducted genome-wide mRNA decay analysis to get further insights into the physiology of Gluconobacter oxydans.

2018-09-21 | GSE103428 | GEO

Complete Genome Sequence of the Industrial Strain Gluconobacter oxydans H24.

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CRISPR/Cpf1-FOKI-induced gene editing in <i>Gluconobacter oxydans</i>.

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