Cloning and Expression of Three New Aazotobacter Vinelandii Genes Closely Related to a Previously Described Gene Family Encoding Mannuronan C-5-epimerases

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1998 Dec 29

PMID 9864314

Citations 26

Authors

B I Svanem

G Skjak-Braek

H Ertesvag

S Valla

Affiliations

Soon will be listed here.

Abstract

The cloning and expression of a family of five modular-type mannuronan C-5-epimerase genes from Azotobacter vinelandii (algE1 to -5) has previously been reported. The corresponding proteins catalyze the Ca2+-dependent polymer-level epimerization of beta-D-mannuronic acid to alpha-L-guluronic acid (G) in the commercially important polysaccharide alginate. Here we report the identification of three additional structurally similar genes, designated algE6, algE7, and algY. All three genes were sequenced and expressed in Escherichia coli. AlgE6 introduced contiguous stretches of G residues into its substrate (G blocks), while AlgE7 acted as both an epimerase and a lyase. The epimerase activity of AlgE7 leads to formation of alginates with both single G residues and G blocks. AlgY did not display epimerase activity, but a hybrid gene in which the 5'-terminal part was exchanged with the corresponding region in algE4 expressed an active epimerase. Southern blot analysis of genomic A. vinelandii DNA, using the 5' part of algE2 as a probe, indicated that all hybridization signals originated from algE1 to -5 or the three new genes reported here.

Citing Articles

Computational modeling of the molecular basis for the calcium-dependence of the mannuronan C-5 epimerase AvAlgE6 from .

Gaardlos M, Lervik A, Samsonov S Comput Struct Biotechnol J. 2023; 21:2188-2196.

PMID: 37013001 PMC: 10066508. DOI: 10.1016/j.csbj.2023.03.021.

Strain Construction and Process Development for Efficient Recombinant Production of Mannuronan C-5 Epimerases in .

Tondervik A, Aune R, Degelmann A, Piontek M, Ertesvag H, Skjak-Braek G Front Plant Sci. 2022; 13:837891.

PMID: 35734252 PMC: 9208277. DOI: 10.3389/fpls.2022.837891.

Mechanistic Basis for Understanding the Dual Activities of the Bifunctional Azotobacter vinelandii Mannuronan C-5-Epimerase and Alginate Lyase AlgE7.

Gaardlos M, Heggeset T, Tondervik A, Teze D, Svensson B, Ertesvag H Appl Environ Microbiol. 2021; 88(3):e0183621.

PMID: 34878812 PMC: 8824271. DOI: 10.1128/AEM.01836-21.

Molecular weight and guluronic/mannuronic ratio of alginate produced by Azotobacter vinelandii at two bioreactor scales under diazotrophic conditions.

Diaz-Barrera A, Sanchez-Rosales F, Padilla-Cordova C, Andler R, Pena C Bioprocess Biosyst Eng. 2021; 44(6):1275-1287.

PMID: 33635396 DOI: 10.1007/s00449-021-02532-8.

Identification of a Pivotal Residue for Determining the Block Structure-Forming Properties of Alginate C-5 Epimerases.

Stanisci A, Tondervik A, Gaardlos M, Lervik A, Skjak-Braek G, Sletta H ACS Omega. 2020; 5(8):4352-4361.

PMID: 32149266 PMC: 7057702. DOI: 10.1021/acsomega.9b04490.

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