Studies on chemiluminescent enzymes from Photobacterium leiognathi.

Mr., Jigar (2013) Studies on chemiluminescent enzymes from Photobacterium leiognathi. [Student Project Report] (Submitted)

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Abstract

Chemiluminescence enzymes present in bioluminescent bacterium, Photobacterium leiognathi has been studied since many years and enzymes such as bacterial luciferase, fluorescent proteins such as lumazine protein or Blue fluorescent protein (BFP) and yellow fluorescent protein (YFP) were found to play a prominent role in the emission and modulation of light in P. leiognathi. The fluorescent proteins such as BFB and YFP were found to act as antenna proteins since these proteins were found to shift the emission wavelength of light generated by the bacterial luciferase enzyme via the bioluminescence resonance energy transfer (BRET). However, these fluorescent proteins, due to their lower yields; didn’t seek much attention as potential fluorophores. In the present study, we discovered that 95% of these fluorescent proteins are released by CFTRI-BIOPHOTO-1 strain of P. leiognathi in the extracellular environment and hence were found to be in plenty. These proteins viz. YFP and BFP were salted out and purified using conventional size exclusion chromatography having % yield of 24.24 having specific activity of 2850.70 RFU/mg and purification fold of 7.32. These fluorescent proteins (FPs) were found to be resistant to protease action, stable at a wide range of pH (2-12) and temperature (upto 100˚C). These FPs are under investigation for their interaction with gold nanoparticles and other proteins for a variety of biosensing applications such as bioluminescence imaging and toxin monitoring.

Item Type:	Student Project Report
Uncontrolled Keywords:	Chemiluminescent Enzymes, P. leiognathi, Fluorescent proteins, BRET
Subjects:	500 Natural Sciences and Mathematics > 07 Life Sciences > 04 Microbiology 600 Technology > 08 Food technology > 16 Nutritive value > 05 Enzymes
Divisions:	Fermentation Technology and Bioengineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	28 Apr 2014 09:49
Last Modified:	28 Apr 2014 09:49
URI:	http://ir.cftri.res.in/id/eprint/11494

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