Artículo
A dye elution method for the quantification of insecticidal crystal proteins from Bacillus thuringiensis and its compatibility with the presence of agro-industrial raw materials and waste products
Fecha de publicación:
03/2024
Editorial:
Oxford University Press
Revista:
Letters in Applied Microbiology
ISSN:
1472-765X
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
The insecticidal crystal proteins produced by Bacillus thuringiensis during sporulation are active ingredients against lepidopteran, dipteran, andcoleopteran insects. Several methods have been reported for their quantification, such as crystal counting, ELISA, and SDS-PAGE/densitometry.One of the major tasks in industrial processes is the analysis of raw material dependency and costs. Thus, the crystal protein quantificationmethod is expected to be compatible with the presence of complex and inexpensive culture medium components. This work presents a reval-idated elution-based method for the quantification of insecticidal crystal proteins produced by the native strain B. thuringiensis RT. To quantifyproteins, a calibration curve was generated by varying the amount of BSA loaded into SDS-PAGE gels. First, SDS-PAGE was performed for qual-ity control of the bioinsecticide. Then, the stained protein band was excised from 10% polyacrylamide gel and the protein-associated dye waseluted with an alcoholic solution of SDS (3% SDS in 50% isopropanol) during 45 min at 95◦C. This protocol was a sensitive procedure to quantifyproteins in the range of 2.0–10.0 μg. As proof of concept, proteins of samples obtained from a complex fermented broth were separated bySDS-PAGE. Then, Cry1 and Cry2 proteins were properly quantified.
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Articulos(PROIMI)
Articulos de PLANTA PILOTO DE PROC.IND.MICROBIOLOGICOS (I)
Articulos de PLANTA PILOTO DE PROC.IND.MICROBIOLOGICOS (I)
Citación
Mentel, Maria Isabel; Loto, Flavia del Valle; Baigori, Mario Domingo; Pera, Licia Maria; A dye elution method for the quantification of insecticidal crystal proteins from Bacillus thuringiensis and its compatibility with the presence of agro-industrial raw materials and waste products; Oxford University Press; Letters in Applied Microbiology; 77; 3; 3-2024; 1-7
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