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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/1532
Title: Purification, characterisation and cloning of a 2S albumin with DNase, RNase and antifungal activities from Putranjiva roxburghii
Authors: Tomar P.P.S.
Chaudhary N.S.
Mishra P.
Gahloth D.
Patel G.K.
Selvakumar P.
Kumar P.
Sharma A.K.
Published in: Applied Biochemistry and Biotechnology
Abstract: The present study reports the characterisation of a novel ?12-kDa heterodimeric protein, designated as putrin, from the seeds of Putranjiva roxburghii. The purification of putrin to homogeneity was accomplished using DEAE-sepharose where protein was unbound, CM-sepharose and Cibacron blue 3GA where it was bound and appeared as single peak on a size-exclusion chromatography column. A 15 % sodium dodecyl sulphate polyacrylamide electrophoresis gel, under reducing condition, demonstrated that putrin is made of two polypeptide chains of approximately 4.5 and 7.5 kDa. Circular dichroism studies demonstrated the helical nature and conformational stability of protein at increasing temperatures. Putrin exhibited both RNase and DNase activities and exerted antifungal activity but possessed relatively weak translation- inhibitory activity in cell-free system. The cloning and sequence analysis revealed a 414 bp open reading frame encoding a preproprotein of 137 amino acid residues. The amino acid sequence comparisons and phylogenetic analysis of putrin showed significant homology to 2S seed storage family proteins. The results demonstrated that putrin belongs to 2S albumin family and exhibits a spectrum of biotechnologically exploitable functions. © 2014 Springer Science+Business Media.
Citation: Applied Biochemistry and Biotechnology(2014), 174(2): 471-482
URI: https://doi.org/10.1007/s12010-014-1078-9
http://repository.iitr.ac.in/handle/123456789/1532
Issue Date: 2014
Publisher: Humana Press Inc.
Keywords: 2S albumins
Antifungal activity
CD studies
Deoxyribonuclease activity
Putranjiva roxburghii
Ribonuclease activity
ISSN: 2732289
Author Scopus IDs: 54906718800
24343212400
57209794009
57208611445
57192590014
36493749300
24432768800
57214355701
Author Affiliations: Tomar, P.P.S., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Chaudhary, N.S., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India, Department of Botany, University of Rajasthan, Jaipur 302004, India
Mishra, P., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Gahloth, D., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Patel, G.K., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Selvakumar, P., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Kumar, P., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Sharma, A.K., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India
Funding Details: Acknowledgements This work was supported by a grant no. IRIS ID No. 2011-15280 from Indian Council of Medical Research, Government of India. CD studies and gel-exclusion chromatography were performed at NMR facility and Macromolecular Crystallographic Unit, respectively, at Institute Instrumentation Centre (IIC), IIT Roorkee. The N-terminal sequencing was performed at the protein sequencing facility of IMTECH, Chandigarh. Authors thank Dr. Partha Roy for allowing and Ms. Rajani Salunke for assisting to use luminometer in their lab. NSC and DG thanks UGC, PPST thanks CSIR, GKP thanks MHRD and PS thanks DBT, Government of India, for financial support.
Corresponding Author: Sharma, A.K.; Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee 247667, India; email: aksbsfbs@yahoo.co.in
Appears in Collections:Journal Publications [BT]

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