The cDNA encoding long-chain fatty-acid alcohol oxidase(FAO) was isolated by using RT-PCR method from Candida cloacae grown in a medium containing oleic acid. The cDNA of fatty-acid alcohol oxidase was cloned into exp...The cDNA encoding long-chain fatty-acid alcohol oxidase(FAO) was isolated by using RT-PCR method from Candida cloacae grown in a medium containing oleic acid. The cDNA of fatty-acid alcohol oxidase was cloned into expression plasmid pET17b under T7 promoter and then transformed into E.coli BL21/DE3. The molecular weight of the expressed protein was estimated to be approximately 64 kD by SDS-PAGE. The expressed protein had a specific catalytic activity of fatty-acid alcohol oxidase and its activity was 1090±116 U/ml medium. Northern blotting revealed that there was high expression of fao mRNA in Candida cloacae yeast grown in a medium containing oleic acid.展开更多
A rapid and sensitive fluorometric method for the enzymatic detection of ethanol using CdSe/ZnS quantum dots （QDs） is proposed. The photoluminescence of QDs is sensitive to H202. This finding leads to a novel approa...A rapid and sensitive fluorometric method for the enzymatic detection of ethanol using CdSe/ZnS quantum dots （QDs） is proposed. The photoluminescence of QDs is sensitive to H202. This finding leads to a novel approach for the determination of ethanol using alcohol oxidase （AOx） which, on oxidation of ethanol, produces H202. The method has higher sensitivity, wider analytical range （0.1-8 mmol/L）, and a lower detection limit （0.05 mmol/L）. The relationship between quenching of the photoluminescence of the QDs and the concentration of ethanol is linear.展开更多
文摘The cDNA encoding long-chain fatty-acid alcohol oxidase(FAO) was isolated by using RT-PCR method from Candida cloacae grown in a medium containing oleic acid. The cDNA of fatty-acid alcohol oxidase was cloned into expression plasmid pET17b under T7 promoter and then transformed into E.coli BL21/DE3. The molecular weight of the expressed protein was estimated to be approximately 64 kD by SDS-PAGE. The expressed protein had a specific catalytic activity of fatty-acid alcohol oxidase and its activity was 1090±116 U/ml medium. Northern blotting revealed that there was high expression of fao mRNA in Candida cloacae yeast grown in a medium containing oleic acid.
基金Project(21005067)supported by the National Natural Science Foundation of ChinaProject(11JJ4015)supported by Natural Science Foundation of Hunan Province,China
文摘A rapid and sensitive fluorometric method for the enzymatic detection of ethanol using CdSe/ZnS quantum dots （QDs） is proposed. The photoluminescence of QDs is sensitive to H202. This finding leads to a novel approach for the determination of ethanol using alcohol oxidase （AOx） which, on oxidation of ethanol, produces H202. The method has higher sensitivity, wider analytical range （0.1-8 mmol/L）, and a lower detection limit （0.05 mmol/L）. The relationship between quenching of the photoluminescence of the QDs and the concentration of ethanol is linear.
