title: Comparative analysis of procoagulant and fibrinogenolytic activity of crude protease fractions of turmeric species.
abstract: Turmeric rhizome is a traditional herbal medicine, which has been widely used as a remedy to stop bleeding on fresh cuts and for wound healing by the rural and tribal population of India.
  
  
  OBJECTIVE
  To validate scientific and therapeutic application of turmeric rhizomes to stop bleeding on fresh cuts and its role in wound healing process.
  
  
  METHODS
  The water extracts of thoroughly scrubbed and washed turmeric rhizomes viz., Curcuma aromatica Salisb., Curcuma longa L., Curcuma caesia Roxb., Curcuma amada Roxb. and Curcuma zedoria (Christm.) Roscoe. were subjected to salting out and dialysis. The dialyzed crude enzyme fractions (CEFs) were assessed for proteolytic activity using casein as substrate and were also confirmed by caseinolytic zymography. Its coagulant activity and fibrinogenolytic activity were assessed using human citrated plasma and fibrinogen, respectively. The type of protease(s) in CEFs was confirmed by inhibition studies using specific protease inhibitors.
  
  
  RESULTS
  The CEFs of C. aromatica, C. longa and C. caesia showed 1.89, 1.21 and 1.07 folds higher proteolytic activity, respectively, compared to papain. In contrast to these, C. amada and C. zedoria exhibited moderate proteolytic activity. CEFs showed low proteolytic activities compared to trypsin. The proteolytic activities of CEFs were confirmed by caseinolytic zymography. The CEFs of C. aromatica, C. longa and C. caesia showed complete hydrolysis of Aα, Bβ and γ subunits of human fibrinogen, while C. amada and C. zedoria showed partial hydrolysis. The CEFs viz., C. aromatica, C. longa, C. caesia, C. amada and C. zedoria exhibited strong procoagulant activity by reducing the human plasma clotting time from 172s (Control) to 66s, 84s 88s, 78s and 90s, respectively. The proteolytic activity of C. aromatica, C. longa, C. caesia and C. amada was inhibited (>82%) by PMSF, suggesting the possible presence of a serine protease(s). However, C. zedoria showed significant inhibition (60%) against IAA and moderate inhibition (30%) against PMSF, indicating the presence of cysteine and serine protease(s).
  
  
  CONCLUSIONS
  The CEFs of turmeric species exhibited strong procoagulant activity associated with fibrinogenolytic activity. This study provides the scientific credence to turmeric in its propensity to stop bleeding and wound healing process practiced by traditional Indian medicine.
authors.affiliation: Department of Biotechnology, Sri Jayachamarajendra College of Engineering, JSS Technical Institutions Campus, Mysuru 570006, Karnataka, India; JSS research foundation, JSS Technical Institutions Campus, Mysuru 570006, Karnataka, India.
authors.forename: B R
authors.initials: BR
authors.lastname: Shivalingu
authors.affiliation: JSS research foundation, JSS Technical Institutions Campus, Mysuru 570006, Karnataka, India.
authors.forename: H K
authors.initials: HK
authors.lastname: Vivek
authors.affiliation: Department of Biotechnology, Sri Jayachamarajendra College of Engineering, JSS Technical Institutions Campus, Mysuru 570006, Karnataka, India; JSS research foundation, JSS Technical Institutions Campus, Mysuru 570006, Karnataka, India.
authors.forename: Zohara
authors.initials: Z
authors.lastname: Nafeesa
authors.affiliation: Department of Studies in Chemistry, University of Mysore, Mysuru 570006, Karnataka, India.
authors.forename: B S
authors.initials: BS
authors.lastname: Priya
authors.affiliation: JSS research foundation, JSS Technical Institutions Campus, Mysuru 570006, Karnataka, India. Electronic address: nanju_chem@yahoo.com.
authors.forename: S Nanjunda
authors.initials: SN
authors.lastname: Swamy
mesh_terms: D000328:Adult; D001777:Blood Coagulation / Q000187:drug effects; D003029:Coagulants / Q000302:isolation & purification / Q000494:pharmacology*; D030024:Curcuma / Q000737:chemistry*; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D007194:India; D008519:Medicine, Traditional; D010447:Peptide Hydrolases / Q000378:metabolism; D010936:Plant Extracts / Q000494:pharmacology*; D027343:Rhizome; D057057:Serine Proteases / Q000378:metabolism; D014945:Wound Healing / Q000187:drug effects
chemical_list: D003029:Coagulants; D010936:Plant Extracts; D005340:Fibrinogen; D010447:Peptide Hydrolases; D057057:Serine Proteases
keywords: Curcuma longa Linn; Fibrinogenolytic; Hemostasis; Pro-coagulant; Serine protease(s)