A study on the Class III Endochitinase of Tamarind (Tamarindus indica) seeds

Sudhanva, M. S. (2010) A study on the Class III Endochitinase of Tamarind (Tamarindus indica) seeds. [Student Project Report]

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Abstract

This Dissertation / Report is the outcome of investigation carried out by the creator(s) / author(s) at the department/division of Central Food Technological Research Institute (CFTRI), Mysore mentioned below in this page.

Item Type:	Student Project Report
Additional Information:	Tamarinin a homolog of Class III chitinases accounts for up to 50% of the total protein from the Tamarind seeds (Tamarindus indica) and belongs to the family 18 glycosyl hydrolases. This 34000 Da protein physiologically serves as the major storage protein, but lacks catalytic activity. Amino‐terminal sequence analyses revealed that Tamarinin is similar to other Class III chitinases. The present investigation entitled “A study on the Class III endochitinase of tamarind (Tamarindus indica) seeds involved purification and characterization of tamarinin the Class III endochitinase from tamarind seeds and delineates the primary structure by cDNA sequence determination. Tamarinin was purified to homogeneity from the alkaline crude extract by chitin bead affinity chromatography. Homogeneity of the purified tamarinin was assess by native page, chitinase zymogram, RP‐HPLC, immunoblot using antibody raised against purified tamarinin and amino‐terminal analysis. Schiff’s staining showed that the purified tamarinin is a glycoprotein. Genomic DNA and total RNA was extracted from young mid matured tamarind seeds. Genomic DNA was used to amplify 759 bp fragment of the tamarinin gene which was further used to design primers for RLM‐RACE. The 5' RACE 150 bp of unknown upstream sequence was deduced. 3' RACE established the 48 bp downstream region of the known tamarinin gene fragment. The deduced protein sequence of tamarinin showed high sequence similarity and shows that all the half cystines and family 18 glycosyl hydrolases signature sequence is conserved. Analysis to predict the signal peptide cleavage showed that the signal peptide is cleaved after 27th amino acid residue which is concurrent with the amino‐terminal analysis of the mature protein. Hydropathy calculations showed that the amino‐terminal and carboxy‐terminal regions of tamarinin are hydrophobic where as the core of the tamarinin is predominantly hydrophilic. Secondary structure prediction showed that tamarinin has 7 α helices and 10 β strands. Since tamarinin is a glycoprotein analysis of the deduced protein sequence N and O glycosylation showed that Asn 130 and Thr 89 are the highly probable N and O glycosylation sites respectively.
Uncontrolled Keywords:	Class III chitinases; Tamarinin; Tamarind seeds; Tamarindus indica
Subjects:	500 Natural Sciences and Mathematics > 04 Chemistry and Allied Sciences > 29 Protein Chemistry 500 Natural Sciences and Mathematics > 10 Plants
Divisions:	Protein Chemistry and Technology
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	14 Jul 2010 09:59
Last Modified:	28 Dec 2011 10:16
URI:	http://ir.cftri.res.in/id/eprint/9556

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