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Titolo:
MOLECULAR-CLONING, EXPRESSION AND EVALUATION OF PHOSPHOHYDROLASES FORPHYTATE-DEGRADING ACTIVITY
Autore:
MOORE E; HELLY VR; CONNEELY OM; WARD PP; POWER RF; HEADON DR;
Indirizzi:
NATL UNIV IRELAND UNIV COLL GALWAY,ALLTECH RES INT GALWAY IRELAND BAYLOR COLL MED,DEPT CELL BIOL HOUSTON TX 77030 NATL UNIV IRELAND UNIV COLL GALWAY,DEPT BIOCHEM,CELL & MOLEC BIOL GRPGALWAY IRELAND
Titolo Testata:
Journal of industrial microbiology
fascicolo: 5, volume: 14, anno: 1995,
pagine: 396 - 402
SICI:
0169-4146(1995)14:5<396:MEAEOP>2.0.ZU;2-G
Fonte:
ISI
Lingua:
ENG
Soggetto:
YEAST ACID-PHOSPHATASE; ASPERGILLUS-NIGER PHYTASE; EXTRACELLULAR PHYTASE; GENE; PHOSPHORUS; FICUUM; GLYCOSYLATION; PURIFICATION; SECRETION; PIGS;
Keywords:
ACID PHOSPHATASE; PHYTASE; ASPERGILLUS; SACCHAROMYCES CEREVISIAE; PHOSPHORUS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
41
Recensione:
Indirizzi per estratti:
Citazione:
E. Moore et al., "MOLECULAR-CLONING, EXPRESSION AND EVALUATION OF PHOSPHOHYDROLASES FORPHYTATE-DEGRADING ACTIVITY", Journal of industrial microbiology, 14(5), 1995, pp. 396-402

Abstract

Four acid phosphatase (phosphomonoesterase E.C.3.1.3.2) genes were cloned by polymerase chain reaction (PCR). These were pho3, pho5 and pho11 from Saccharomyces cerevisiae and the gene for a phosphate-respressible acid phosphatase from Aspergillus niger. The individual genes were subcloned into an A. oryzae expression vector downstream from a starch-inducible alpha-amylase promoter and the resulting expression constructs were transformed into a mutant strain of A. oryzae, AO7. Southern hybridization analysis confirmed that the acid phosphatase genes hadbeen integrated into the host genome with estimates of integrated copy numbers ranging from 2 to 20 for individual transformants. Northern hybridization analysis of total RNA from individual transformants revealed the presence of a single transcript of the expected size of 1.8 kb. Production of recombinant protein was induced by the addition of 30g L(-1) of soluble starch in the fermentation media. Active acid phosphatases, not present in control cultures, were detected in the supernatant fractions of transformant cultures by acid phosphatase activity staining of non-denaturing polyacrylamide gels. The ability of the recombinant acid phosphatases to hydrolyze phytate was assessed by referenced phytase (myoinositol hexakisphosphate phosphohydrolase E.C. 3.1.3.8) activity assay procedures. A two- to six-fold increase in phytase activity was measured in transformants compared to control, untransformed A. oryzae. Sufficient quantities of A. niger and pho5 recombinant acid phosphatases were generated from large-scale fermentations to assess the efficacy of these enzymes as phytate-degrading enzymes when included in poultry diets. Data indicated an increase in available phosphorus of 1 g kg(-1) obtained with yeast acid phosphatase and A. niger acid phosphatase representing 40% utilization of unavailable dietary Pcompared to 48% utilization for commercial phytase.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/09/20 alle ore 13:12:53