Construction of a Xylanase-Producing Strain of Brevibacterium lactofermentum by Stable Integration of an Engineered xysA Gene from Streptomyces halstedii JM8

Sirin A I Adham, Ana B. Campelo, Angelina Ramos, José A. Gil

Research output: Contribution to journalArticle

12 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)5425-5430
Number of pages6
JournalApplied and Environmental Microbiology
Volume67
Issue number12
DOIs
Publication statusPublished - Dec 2001

Fingerprint

Streptomyces halstedii
Brevibacterium
Corynebacterium glutamicum
Streptomyces
xylanases
gene
Genes
genes
nitrilotriacetic acid
Nitrilotriacetic Acid
Chromosomes, Human, 4-5
antibiotic resistance
Microbial Drug Resistance
plasmid
Cell Division
antibiotics
Sepharose
agarose
cell division
chromosome

ASJC Scopus subject areas

  • Biotechnology
  • Environmental Science(all)
  • Microbiology

Cite this

Construction of a Xylanase-Producing Strain of Brevibacterium lactofermentum by Stable Integration of an Engineered xysA Gene from Streptomyces halstedii JM8. / Adham, Sirin A I; Campelo, Ana B.; Ramos, Angelina; Gil, José A.

In: Applied and Environmental Microbiology, Vol. 67, No. 12, 12.2001, p. 5425-5430.

Research output: Contribution to journalArticle

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