A simple method for separation of the protein crystals from Bacillus thuringiensis using carboxymethyl cellulose column chromatography

• Published in: Journal of microbiological methods Vol. 19; no. 2; pp. 103 - 110
• Main Authors: MURTY, M. G, SRINIVAS, G, ROOP SINGH BORA, VAITHILINGAM SEKAR
• Format: Journal Article
• Language: English
• Published: Shannon Elsevier Science 01.01.1994
• Subjects: Bacillus thuringiensis; Bacteriological methods and techniques used in bacteriology; Bacteriology; Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Microbiology; Purification; Bacillales; Bacillus thuringiensis; Parasporal crystal; Bacteria; Method; Bacillaceae; Column chromatography; Spores
• ISSN: 0167-7012; 1872-8359
• DOI: 10.1016/0167-7012(94)90041-8

A simple and a rapid technique to separate intact crystals from spores and cell debris of Bacillus thuringiensis using a carboxymethyl cellulose (CM-cellulose) column is described. B. thuringiensis subsp. kurstaki, a toxic strain to lepidopteran insects that produces small bipyramidal crystals, and B. thuringiensis subsp. yunnanensis, a nontoxic strain producing unusually large bipyramidal and spindle-shaped crystals were used as test organisms. Spores and cell debris were washed in sodium acetate and sodium phosphate buffers and the crystals bound to the CM-cellulose were eluted with Tris-EDTA buffer, regardless of their size and shape. Light microscopy and sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the purity of the crystal was >98%. The percent recoveries of crystals from B. thuringiensis subsp. kurstaki and B. thuringiensis subsp. yunnanensis were approximately 14 and 25, respectively.