NDLI: Deletion of pyruvate decarboxylase by a new method for efficient markerless gene deletions in Gluconobacter oxydans

Content Provider	Springer Nature Link
Author	Peters, Björn Junker, Anja Brauer, Katharina Mühlthaler, Bernadette Kostner, David Mientus, Markus Liebl, Wolfgang Ehrenreich, Armin
Copyright Year	2012
Abstract	Gluconobacter oxydans, a biotechnologically relevant species which incompletely oxidizes a large variety of carbohydrates, alcohols, and related compounds, contains a gene for pyruvate decarboxylase (PDC). This enzyme is found only in very few species of bacteria where it is normally involved in anaerobic ethanol formation via acetaldehyde. In order to clarify the role of PDC in the strictly oxidative metabolism of acetic acid bacteria, we developed a markerless in-frame deletion system for strain G. oxydans 621H which uses 5-fluorouracil together with a plasmid-encoded uracil phosphoribosyltransferase as counter selection method and used this technique to delete the PDC gene (GOX1081) of G. oxydans 621H. The PDC deletion mutant accumulated large amounts of pyruvate but almost no acetate during growth on d-mannitol, d-fructose or in the presence of l-lactate. This suggested that in G. oxydans acetate formation occurs by decarboxylation of pyruvate and subsequent oxidation of acetaldehyde to acetate. This observation and the efficiency of the markerless deletion system were confirmed by constructing deletion mutants of two acetaldehyde dehydrogenases (GOX1122 and GOX2018) and of the acetyl-CoA-synthetase (GOX0412). Acetate formation during growth of these mutants on mannitol did not differ significantly from the wild-type strain.
Starting Page	2521
Ending Page	2530
Page Count	10
File Format	PDF
ISSN	01757598
Journal	Applied Microbiology and Biotechnology
Volume Number	97
Issue Number	6
e-ISSN	14320614
Language	English
Publisher	Springer-Verlag
Publisher Date	2012-09-01
Publisher Place	Berlin, Heidelberg
Access Restriction	Subscribed
Subject Keyword	Gluconobacter oxydans Acetic acid bacteria Uracil phosphoribosyltransferase Markerless deletion Pyruvate decarboxylase Acetate metabolism Biotechnology Microbiology Microbial Genetics and Genomics
Content Type	Text
Resource Type	Article
Subject	Medicine Applied Microbiology and Biotechnology Biotechnology

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

Importance of codB for new codA-based markerless gene deletion in Gluconobacter strains

Characterization of membrane-bound dehydrogenases of Gluconobacter oxydans 621H using a new system for their functional expression

Asymmetric reduction of diketones by two Gluconobacter oxydans oxidoreductases

Vinyl ketone reduction by three distinct Gluconobacter oxydans 621H enzymes

Analysis of aldehyde reductases from Gluconobacter oxydans 621H

Characterization of enzymes involved in the central metabolism of Gluconobacter oxydans

Identification of mannitol as compatible solute in Gluconobacter oxydans

The consequence of an additional NADH dehydrogenase paralog on the growth of Gluconobacter oxydans DSM3504

Draft genome sequence of Gluconobacter thailandicus NBRC 3257

Deletion of pyruvate decarboxylase by a new method for efficient markerless gene deletions in Gluconobacter oxydans

Similar Documents

Importance of codB for new codA-based markerless gene deletion in Gluconobacter strains

Characterization of membrane-bound dehydrogenases of Gluconobacter oxydans 621H using a new system for their functional expression

Asymmetric reduction of diketones by two Gluconobacter oxydans oxidoreductases

Vinyl ketone reduction by three distinct Gluconobacter oxydans 621H enzymes

Analysis of aldehyde reductases from Gluconobacter oxydans 621H

Characterization of enzymes involved in the central metabolism of Gluconobacter oxydans

Identification of mannitol as compatible solute in Gluconobacter oxydans

The consequence of an additional NADH dehydrogenase paralog on the growth of Gluconobacter oxydans DSM3504

Draft genome sequence of Gluconobacter thailandicus NBRC 3257

Deletion of pyruvate decarboxylase by a new method for efficient markerless gene deletions in Gluconobacter oxydans