Easy and Non-Expensive Method to Increase the Production, Enhance the Stability and Improve the Purification of Cry1Ab Crystals

Rosalía Contreras¹, Christian Hernández¹, Manuel Acosta¹, Viviana Pitones², Gretel Mendoza³ & Jorge Olmos^1*

¹Molecular Microbiology Laboratory, Department of Marine Biotechnology, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), México
²Faculty of Dentistry, Universidad Autónoma de Baja California, México
³Academic Unit of Biological Sciences, Universidad Autónoma de Zacatecas, México

Dr. Jorge Olmos, Molecular Microbiology Laboratory, Department of Marine Biotechnology, Centro de Investigación Científica y de Educación Superior de Ensenada (CICESE), México.

Keywords: Bacillus Thuringiensis; Cry1Ab; PH Stability; Filtration; Purification

Abstract

We were able to increase Bacillus thuringiensis Cry1Ab crystals production and enhance its stability by using C3.5:N4.5 nutrients ratio. Crystal size was conveniently reduced from ≥1.0 to ≤ 0.5 μm by increasing culture aeration conditions, in order to facilitate their purification by a simple filtration procedure The recombinant Cry1Ab protein was produced 10 times more using the C3.5:N4.5 ratio, with respect to the amount obtained using the C7:N1 and C1:N7 proportions. No gradients neither solvent treatments were needed to get a highly purified crystals with insecticidal activity. No spores were observed under the microscope using malachite green stain.

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