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Titolo:
Efficient removal of sulfide following integration of multiple copies of the sulfide-quinone oxidoreductase gene (sqr) into the Escherichia coli chromosome
Autore:
Shibata, H; Takahashi, M; Yamaguchi, I; Kobayashi, S;
Indirizzi:
Meiji Univ, Lab Anim Environm, Tama Ku, Kawasaki, Kanagawa 2148571, Japan Meiji Univ Kawasaki Kanagawa Japan 2148571 asaki, Kanagawa 2148571, Japan
Titolo Testata:
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
fascicolo: 5, volume: 91, anno: 2001,
pagine: 493 - 499
SICI:
1389-1723(200105)91:5<493:EROSFI>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
PARACOCCUS-DENITRIFICANS GB17; RHODOBACTER-CAPSULATUS; CYTOCHROME-B562-O COMPLEX; OSCILLATORIA-LIMNETICA; CHROMATIUM-VINOSUM; REDUCTASE-ACTIVITY; FLAVOCYTOCHROME-C; DEHYDROGENASE; PURIFICATION; EXPRESSION;
Keywords:
sulfide oxidation; sulfide-quinone oxidoreductase; homologous recombination; chromosomal integration; recombinant Escherichia coli;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
32
Recensione:
Indirizzi per estratti:
Indirizzo: Kobayashi, S Meiji Univ, Lab Anim Environm, Tama Ku, 1-1-1 Higashimita, Kawasaki, Kanagawa 2148571, Japan Meiji Univ 1-1-1 Higashimita Kawasaki Kanagawa Japan 2148571
Citazione:
H. Shibata et al., "Efficient removal of sulfide following integration of multiple copies of the sulfide-quinone oxidoreductase gene (sqr) into the Escherichia coli chromosome", J BIOSCI BI, 91(5), 2001, pp. 493-499

Abstract

For the oxidation and removal of hydrogen sulfide, which causes an offensive odor from the contents of animal intestines, recombinant strains of Escherichia coli were constructed. The sulfide-quinone oxidoreductase gene (sqr) from Rhodobacter capsulatus was integrated in low copy numbers into the chromosome of Escherichia coli W3110. Multiple copies of sqr on plasmids were also delivered into the cytoplasm of the same strain. The sqr genes were homologously transducted onto the chromosomal lacZ region and their existence there was verified by Southern blot analysis. Sulfide oxidation in a chemical medium effectively increased for the recombinant strains which carried 2 similar to3 copies of sqr under the control of the lac or tac promoter in the chromosome, and also for strains which carried 10 copies of sqr under the control of the lac or tac promoter on plasmids. In both types of recombinant, the tac promoter was more effective for SQR expression than the lac promoter. Construction of a recombinant with 3 copies of sqr under the control of the tac promoter in the chromosome was unsuccessful. In recombinants with SQR activity lower than 700 nmol/mg cell protein/min, oxygen consumption increased proportionally to SQR activity. An elevation in SQR activity in this range resulted in an increase in oxygen consumption and a decrease in sulfide concentration. When the recombinant cells were cultured until the 160th generation, WL2, WL3 and WT2, which carried 2, 3 and 2 copies of sqr in the chromosome, respectively, retained SQR activity similar to that of the first generation. For WL300 and WT20 which carried multi-copies of sqr in plasmids SQR activity was undetectable. The recombinant with 2 copiesof sqr in the chromosome regulated by the tac promoter was most suitable for sulfide oxidation and growth of the cells.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 31/03/20 alle ore 10:24:34