A Different Method of Measuring and Detecting Mono-and Dioxygenase Activities

Zazueta-Sandoval, Roberto; Novoa, Vanesa Zazueta; Jiménez, Hortencia Silva; Ortiz, Roberto Cabrera

doi:10.1007/978-1-4612-0057-4_60

Cited by 3 publications

(5 citation statements)

References 9 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One unit of enzyme activity was defined as the amount of enzyme that leads to the production of 1 μmol H 2 O 2 per minute at 25 °C. FAO specific activity was expressed as units per mg of protein Oxygenase activity was assayed as described previously by Zazueta-Sandoval et al (2003).…”

Section: Preparation Of Cell Free Extractsmentioning

confidence: 99%

Peroxidases in YR-1 strain ofMucor circinelloides a potential bioremediator of petroleum-contaminated soils

2008

Self Cite

View full text Add to dashboard Cite

We have studied the induction and subcellular localisation of peroxidase activity present in cell-free extracts of YR-1 strain of Mucor circinelloides in relation with different growth conditions, particularly different carbon sources; the results obtained indicated that the peroxidase expression could be an inducible mechanism depending on the carbon source used. Peroxidase was induced in the presence of hydrocarbons or methanol meanwhile in glucose no activity was founded. On zymograms of peroxidase activity after polyacrylamide gel-electrophoresis two different activity bands were revealed, one of them (major band) shows different relative mobility depending on the aliphatic hydrocarbon chain length used as carbon source, indicating the possibility that could exist different iso-forms of the peroxidase activity. In an immunological approach using a polyclonal antibody against commercial peroxidase, the result shows that the antibody recognised the same two bands revealed in zymogram indicating that activity bands possesses a similar epitope in their structure. On SDS-PAGE loading supernatants obtained by high-speed centrifugation from cell-free extracts of YR-1 grown in glucose, n-decane or methanol, were detected different prominent protein bands at (in kDa): 64 and 94 (glucose), 27 and 46 (n-decane), and 28 and 47 (n-hexadecane) in comparison with a 55 kD band of horseradish peroxidase used as a control. These protein-bands could be the putative subunits of each iso-form in the native protein. Preliminary studies about the intracellular localisation of the enzyme using isopicnic-sucrose gradients suggest that it is located in the peroxisomal fraction.

show abstract

Section: Preparation Of Cell Free Extractsmentioning

confidence: 99%

Peroxidases in YR-1 strain ofMucor circinelloides a potential bioremediator of petroleum-contaminated soils

2008

Self Cite

View full text Add to dashboard Cite

show abstract

“…In addition the results show the dependence of enzyme activity with temperature. Zazueta-Sandoval et al (2003) did not observe effect of incubation temperature at 28 or 37 • C, for the unknown strain they used. purified and characterized a benzoateparahydroxylase of A. niger.…”

Section: The Effect Of Temperature On Oxygenase(s) Activitymentioning

confidence: 71%

“…Oxygenase activity assays were performed, in cell free crude enzyme extracts. Two methods were used: oxygen uptake rate (oxymetry), based on a method described by and a modified spectrophotometric method described by Zazueta-Sandoval et al (2003).…”

Section: Analytical Proceduresmentioning

confidence: 99%

“…Enzyme activity was calculated subtracting reference slope (absorbance at 460 nm vs time) from sample slope and interpolating this value on a peroxidase activity calibration curve made with different concentrations of hydrogen peroxide as substrate of the enzyme in the presence of odianisidine (Zazueta-Sandoval et al, 2003). This method is based in the characteristics of ortodianisidine (orto-DNS) which is a chromogenic acceptor of oxygen and in the action of oxygenase on hydrocarbon molecule and peroxidase (POD) on hydrogen peroxide, accordingly with the next two net reactions, using hexadecane as example of substrate for a monooxygenase (MOX):…”

Section: Analytical Proceduresmentioning

confidence: 99%

“…Oxygenase(s) substrate specificity was analyzed by the spectrophotometric method (Zazueta et al, 2003).…”

Section: Characterization Of the Oxygenase(s)mentioning

confidence: 99%

See 2 more Smart Citations

Producción de oxigenasas de hidrocarburos por Aspergillus niger por fermentación en estado sólido

Flores¹,

Rojas²

View full text Add to dashboard Cite

en FES no tiene una especificidad diferente para hidrocarburos alifáticos (lineales o ramificados). Sin embargo, la especificidad para compuestos aromáticos se incrementa con Resumen la complejidad del anillo. La mayor especificidad se presenta con tolueno y pireno. La actividad del extracto enzimático de FEL no presenta un patrón determinado y muestra actividad tanto con aromáticos como con alifáticos.Se evaluó la actividad oxigenasa en extractos enzimáticos de biomasa obtenida tanto en FEL como en FES, encontrándose que la actividad es significativamente diferente para la biomasa de ambos sistemas de fermentación. La actividad del extracto enzimático de FEL es por lo menos 7 veces menor que la actividad en FES.Palabras clave: oxigenasas, fermentación en estado sólido, hidrocarburos aromáticos, hexadecano, soporte de poliuretano, Aspergillus niger.

show abstract

Actividades dihidrodiol-deshidrogenasa en Mucor circinelloides YR-1 aislada de suelo contaminado con petróleo

Morales¹,

Durón-Castellanos²,

Flores-Martínez³

et al. 2010

Acta Universitaria

View full text Add to dashboard Cite

Después de crecer células de la cepa YR-1 de Mucor circinelloides en naftaleno como única fuente de carbono, fue revelada la presencia de cinco actividades: 1, 2-dihidroxi-1, 2-dihidro-naftalendiol deshidrogenasa mediante zimogramas electroforéticos, utilizando (+)-cis-1(R), 2(s)-dihidroxi-1, 2-dihidronaftaleno como sustrato y NADP+ como aceptor de electrones. Las bandas NDD1, NDD3 y NDD5 que mostraron una mayor intensidad en los zimogramas y por su movilidad electroforética, fueron purificadas a homogeneidad por pasos secuenciales de electroforesis-electroelución para conocer tanto el peso molecular, como el número de subunidades que conforman a cada banda de actividad. Los resultados mostraron pesos moleculares de 98.64 kDa, 48.87 kDa y 102.45 kDa, para cada subunidad respectiva. Se concluye que las actividades no son isoformas conformadas por la misma subunidad proteica, las enzimas nativas son homopoliméricas y no contienen subunidades protoméricas compartidas.

show abstract

A Different Method of Measuring and Detecting Mono-and Dioxygenase Activities

Cited by 3 publications

References 9 publications

Peroxidases in YR-1 strain ofMucor circinelloides a potential bioremediator of petroleum-contaminated soils

Peroxidases in YR-1 strain ofMucor circinelloides a potential bioremediator of petroleum-contaminated soils

Producción de oxigenasas de hidrocarburos por Aspergillus niger por fermentación en estado sólido

Actividades dihidrodiol-deshidrogenasa en Mucor circinelloides YR-1 aislada de suelo contaminado con petróleo

Contact Info

Product

Resources

About