Purification and properties of cysteine protease from rhizomes of Curcuma longa (Linn.).
BACKGROUND
Turmeric rhizome (Curcuma domestica Linn.) contains proteases and has proteolytic activity. Curcumin from turmeric rhizomes has been used for healing many ailments, including cancer. The purpose of this study was to purify turmeric protease and to research their biochemical characteristics [corrected].


RESULTS
Cysteine protease from C. domestica has been purified to homogeneity using acetone precipitation followed by preparatory native polyacrylamide gel electrophoresis (PAGE). This protocol resulted in six fold purification with 28% final recovery. The purified turmeric protease showed a prominent single peak and band on high-performance liquid chromatography and sodium dodecyl sulfate-PAGE, respectively, and an estimated molecular weight of 43 KDa, and exhibited optimal activity between 37 and 60 degrees C. The protease activity of the turmeric protease was significantly inhibited by iodoacetic acid. The turmeric protease had higher alanine and glutamate content and cleaved synthetic peptides N-Cbz-Ile-Pro and N-Cbz-Phe-Leu in a time-dependent manner. Peptide mass fingerprint using matrix-assisted laser desorption/ionization-time of flight mass spectroscopy revealed peptide matches to proteasome subunit alpha type 3 of Oryza sativa ssp. japonica (Rice). The turmeric protease showed antifungal activity at 10 microg mL(-1) towards pathogens Pythium aphanidermatum, Trichoderma viride and Fusarium sp.


CONCLUSIONS
Cysteine addition significantly activated turmeric protease. The protease inhibition test suggested that turmeric protease belonged to the cysteine type. The biochemical characteristics of turmeric protease described in this paper can provide useful information for potential end uses of turmeric protease for pharmaceutical industry applications such as therapeutics.
Nagarathnam
Radhakrishnan
R
Centre for Advanced studies in Botany, Guindy Campus, University of Madras, Chennai 600025, Tamil Nadu, India.
Rengasamy
Anandhan
A
Balasubramanian
Rudresan
R
D000409:Alanine / Q000032:analysis; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D030024:Curcuma / Q000201:enzymology*; D057056:Cysteine Proteases / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005658:Fungi / Q000187:drug effects*; D018698:Glutamic Acid / Q000032:analysis; D019807:Iodoacetic Acid; D013058:Mass Spectrometry; D012275:Oryza; D010455:Peptides / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry / Q000494:pharmacology*; D010940:Plant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D011480:Protease Inhibitors; D046988:Proteasome Endopeptidase Complex; D027343:Rhizome / Q000201:enzymology*
D000935:Antifungal Agents; D010455:Peptides; D010936:Plant Extracts; D010940:Plant Proteins; D011480:Protease Inhibitors; D018698:Glutamic Acid; D057056:Cysteine Proteases; D046988:Proteasome Endopeptidase Complex; D000409:Alanine; D019807:Iodoacetic Acid