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PMID: 11722888 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Construction of a xylanase-producing strain of Brevibacterium lactofermentum by stable integration of an engineered xysA gene from Streptomyces halstedii JM8.

Applied and environmental microbiology ·Vol. 67 ·No. 12 ·2001-12-00 ·Pages 5425-30

Adham SA, Campelo AB, Ramos A, Gil JA

Abstract

A xylanolytic strain of Brevibacterium lactofermentum containing the Streptomyces halstedii His-tagged xysA gene was generated. The new strain contains DNA derived from S. halstedii, expresses xylanolytic activity, and was obtained by an integrative process mediated by a conjugative plasmid targeted to a dispensable chromosomal region located downstream from the essential cell division gene ftsZ. The His-tagged Xys1 enzyme was constitutively expressed under the control of the kan promoter from Tn5 and was easily purified by use of Ni-nitrilotriacetic acid-agarose. The new strain is stable for more than 200 generations, lacks any known antibiotic resistance gene, and does not need any selective pressure to maintain the integrated gene. This strategy can be used to integrate any gene into the B. lactofermentum chromosome and to maintain it stably without the use of antibiotics for selection.

MeSH Terms

Bacterial Proteins Brevibacterium/enzymology,genetics Conjugation, Genetic Plasmids/genetics Recombination, Genetic Streptomyces/enzymology,genetics Xylan Endo-1,3-beta-Xylosidase Xylosidases/biosynthesis,genetics,metabolism

Chemicals

Bacterial Proteins Xylosidases Xylan Endo-1,3-beta-Xylosidase Xys1 protein, Streptomyces halstedii

Authors & Affiliations

4 authors, click to expand affiliations / ORCID

Adham S A

Departamento de Ecología, Genética y Microbiología, Area de Microbiología, Facultad de Biología, Universidad de León, Spain.

Campelo A B

Ramos A

Gil J A

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Article Info

Journal

Applied and environmental microbiology

Abbr.

Appl Environ Microbiol

ISSN

0099-2240

DOI

10.1128/AEM.67.12.5425-5430.2001

Published

2001-12-00

Pages

5425-30

Language

English

Region

United States

NLM ID

7605801

PMCID

PMC93325

Subset

External Links

PubMed source DOI: 10.1128/AEM.67.12.5425-5430.2001 PMC Full Text View journal details

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Construction of a xylanase-producing strain of Brevibacterium lactofermentum by stable integration of an engineered xysA gene from Streptomyces halstedii JM8.

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