title: Purification, characterisation and cloning of a 2S albumin with DNase, RNase and antifungal activities from Putranjiva roxburghii.
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.
authors: Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, 247667, India.
authors: Prabhat Pratap Singh
authors: PP
authors: Tomar
authors: Navneet S
authors: NS
authors: Chaudhary
authors: Pushkar
authors: P
authors: Priyadarshi
authors: Pushkar
authors: P
authors: Mishra
authors: Deepankar
authors: D
authors: Gahloth
authors: Girijesh Kumar
authors: GK
authors: Patel
authors: Purushotham
authors: P
authors: Selvakumar
authors: Pranav
authors: P
authors: Kumar
authors: Ashwani Kumar
authors: AK
authors: Sharma
mesh_terms: D000418:Albumins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000494:pharmacology*; D001483:Base Sequence; D002850:Chromatography, Gel; D002942:Circular Dichroism; D003001:Cloning, Molecular; D017931:DNA Primers; D003851:Deoxyribonucleases / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D019684:Magnoliopsida / Q000737:chemistry*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D012260:Ribonucleases / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid
chemical_list: D000418:Albumins; D000935:Antifungal Agents; D017931:DNA Primers; D003851:Deoxyribonucleases; D012260:Ribonucleases