The Improved Method for Isolation of Photochrome Transmembrane Protein Bacteriorhodopsin from Purple Membranes of Halobacterium Halobacterium Halobium ET 1001

Oleg Mosin, Ignat Ignatov

Abstract


We proposed the improved method for isolation of photochrome transmembraine protein bacteriorhodopsin (output 5 mg from 100 g of wet biomass) capable to transform light energy to electrochemical energy of generated protons H+ and АТP. The protein was isolated from purple membranes of photo-organotrophic halobacterium Halobacterium halobium ET 1001 by cellular autolysis by distilled water, processing of bacterial biomass by ultrasound at 22 KHz, alcohol extraction of low and high-weight molecular impurities, cellular RNA, carotenoids and lipids, solubilization with 0.5% (w/v) SDS-Na, fractionation by MeOH and column gel permeation chromatography (GPC) of the final protein on Sephadex G-200 with 0.1% (w/v) SDS-Na and 2.5 mM ETDA. The homogeneity of the isolated BR was proved by combination of preparative and analytical methods including elecrtophoresis in 12.5% (w/v) PAAG with 0.1% (w/v) SDS-Na and regeneration of apomembranes with 13-trans-retinal.

Keywords: Halobacterium halobium, purple membranes, bacteriorhodopsin, biosynthesis, biomolecular electronics


Full Text: PDF
Download the IISTE publication guideline!

To list your conference here. Please contact the administrator of this platform.

Paper submission email: JMPB@iiste.org

ISSN 2422-8427

Please add our address "contact@iiste.org" into your email contact list.

This journal follows ISO 9001 management standard and licensed under a Creative Commons Attribution 3.0 License.

Copyright © www.iiste.org