Construction of L-lysine-overproducing strains of Brevibacterium lactofermentum by targeted disruption of the hom and thrB genes

Fernández-González, Cristina; Gil, José A.; Mateos, Luís M.; Schwarzer, A.; Schäfer, Andreas; Kalinowski, Jörn; Pühler, Alfred; Martı́n, Juan F.

doi:10.1007/s002530050860

Cited by 16 publications

(23 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Media were supplemented with kanamycin (12.5 g/ml for C. glutamicum and 50 g/ml for E. coli), ampicillin (100 g/ml), or chloramphenicol (20 g/ml) when necessary. Mobilizable plasmids were transferred by conjugation from E. coli S17-1 (donor strain) to the indicated recipient strains of C. glutamicum (21). All reagents were obtained from commercial sources.…”

Section: Methodsmentioning

confidence: 99%

Efflux Permease CgAcr3-1 of Corynebacterium glutamicum Is an Arsenite-specific Antiporter

Villadangos

Gil

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

Section: Methodsmentioning

confidence: 99%

Efflux Permease CgAcr3-1 of Corynebacterium glutamicum Is an Arsenite-specific Antiporter

Villadangos

Gil

et al. 2012

Journal of Biological Chemistry

View full text Add to dashboard Cite

show abstract

“…Expression of both operons is induced by arsenite. Their repressors, CgArsR1 and CgArsR2, which are 66% identical, both have three cysteine residues, two N-terminal residues (Cys 15 and Cys 16 ) that have no counterpart in any of the other characterized members of the ArsR/SmtB family and a third cysteine residue (Cys 55 ) that is near but not within the putative DNA binding domain of the regulatory proteins. The positions of these cysteine residues in the two CgArsRs resemble those of CadC but appear to have arisen independently.…”

mentioning

confidence: 99%

Evolution of Metal(loid) Binding Sites in Transcriptional Regulators

Ordóñez

Thiyagarajan

Cook

et al. 2008

Journal of Biological Chemistry

View full text Add to dashboard Cite

Arsenic, a toxic metalloid, is currently and has always been ranked first on the Superfund List of Hazardous Substances (available on the World Wide Web), in part because of its environmental ubiquity. As a consequence, nearly all organisms have genes that confer resistance to arsenic. Environmental arsenic is sensed by members of the ArsR/SmtB family of metalloregulatory proteins (1-3). These winged helix repressor proteins specifically bind to arsenic and other toxic metals. Consequently, they control expression of genes involved in arsenic biotransformation and efflux. For example, the ArsR repressor encoded by Escherichia coli plasmid R773 binds to the promoter region of its respective ars operon in the absence of As(III) or Sb(III) (4). This homodimeric repressor has the sequence Cys 32 -Val-Cys 34 -Asp-Leu-Cys 37 in the DNA binding domain of each monomer (5, 6). The three sulfur thiolates of the cysteine residues form a very specific three-coordinate binding site for the trivalent metalloids As(III) and Sb(III). Binding of metalloids to R773 ArsR is presumed to induce a conformational change, leading to dissociation from the DNA and hence derepression. The Staphylococcus aureus CadC is a Cd(II)/Pb(II)/Zn(II)-responsive member of the ArsR/SmtB family that has four cysteine residues in the inducer binding domain (7). Of these four cysteine residues, two come from one subunit, whereas the other two come from the other subunit of the homodimer (8, 9). The position of this metal binding site in CadC is congruent to that of the R773 ArsR but is formed between two monomers. CadC also has a second type of metal binding site (DXHX 10 HX 2 E) for Zn(II) at the dimer interface that is not a regulatory site. This site, however, is identical to the regulatory Zn(II) site of SmtB from Synechococcus PCC 7942 (10). Another member of the ArsR/SmtB family is the ArsR from Acidithiobacillus ferrooxidans (AfArsR), 3 which has three cysteine residues (Cys 95 , Cys 96 , and Cys 102 ) at the dimer interface rather than in or near the DNA binding domain (11). These three cysteine residues form a three-coordinate or S 3 binding site for trivalent metalloids (12). Although both the As(III) binding site of AfArsR and the Zn(II) binding site of SmtB are at the C-terminal dimerization domain, the former is formed of three cysteine residues within a single subunit (two sites per dimer), whereas the latter is formed by four residues, two residues from one monomer and two from the other. Thus, metal(loid) binding sites appear to arise by convergent evolution, even in homologous proteins.

show abstract

“…This plasmid cannot replicate in B. lactofermentum but can be transferred by conjugation from E. coli S17-1. This plasmid has been used to interrupt several genes in B. lactofermentum or Corynebacterium glutamicum (4,8,22).…”

Section: Resultsmentioning

confidence: 99%

“…B. lactofermentum R31 was used as the recipient strain. Conjugation between E. coli and B. lactofermentum was performed as described by Fernández-González et al (4). For direct selection of antibiotic-resistant transconjugants, tryptic soy agar (TSA) plates (21) were supplemented with kanamycin at a final concentration of 30 g ml Ϫ1 .…”

Section: Methodsmentioning

confidence: 99%

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

Adham

Campelo

Ramos

et al. 2001

Appl Environ Microbiol

View full text Add to dashboard Cite

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.

show abstract

Construction of L-lysine-overproducing strains of Brevibacterium lactofermentum by targeted disruption of the hom and thrB genes

Cited by 16 publications

References 20 publications

Efflux Permease CgAcr3-1 of Corynebacterium glutamicum Is an Arsenite-specific Antiporter

Efflux Permease CgAcr3-1 of Corynebacterium glutamicum Is an Arsenite-specific Antiporter

Evolution of Metal(loid) Binding Sites in Transcriptional Regulators

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

Contact Info

Product

Resources

About