EXPRESSION AND ENZYMATIC PROPERTIES OF A UNIQUE RECOMBINANT ANTICOAGULANT AND FIBRINOLYTIC ENZYME FROM ACINETOBACTER BAUMANNII TU04
Journal Title: International Journal of Pharmacy and Pharmaceutical Sciences - Year 2015, Vol 7, Issue 12
Abstract
Objective: The objective of this research is to clone and express a new fibrinolytic enzyme encoding serine protease gene in Escherichia coli thus, characterize such purified recombinant.Methods: The recombinant clone was successfully expressed in Lemo21 system and purified using immobilized nickel cation affinity chromatography on a His•bind resin®, followed by ammonium sulfate precipitation and protein filtration in combination. General properties of the purified enzyme were investigated, including the molecular weight, effects of inhibitors and metal ions, substrate specificity, amylolytic activity, fibrinolytic activity and effect of anticoagulant activity in-vitro.Results: The recombinant clone was expressed in Lemo21 system in the cytoplasm in a soluble and active form. The resulting enzyme, SERpro was purified to homogeneity with a purification of 19.35-fold and recovery yield of 4.85%. The enzyme exhibited maximal activity at 37 °C and at pH7.4, respectively. The molecular weight of the purified enzyme was 82 kDa, determined using sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The fibrinogenolysis peptide sequence analysis revealed that SERpro degraded Bβ chain of Fibrin at a much lower rate but cleaved Aα and γ-chain extensively. The enzyme was activated by metal ions such as Mg2+, Fe3+and Zn2+, and was inhibited strongly by PMSF. The clotting time of human blood serum in the presence of 1U SERpro reached a relative partial thromboplastin time of 13.9% with a 1.14-fold increase.Conclusion: The study deduced SERpro as a new protease with anti-thrombotic activity from Acinetobacter baumannii TU04.Â
Authors and Affiliations
Renuka Krishnan, Chun Shiong Chong, Kian Mau Goh, Firdaus Abdul Wahab, Haryati Jamaluddin
EFFECT OF VITAMIN C ON mRNA EXPRESSION OF BMPs DURING FRACTURE HEALING IN THE FEMUR CALLUS OF SPRAGUE-DAWLEY FEMALE RATS
Fracture reparation is a complex physiological process. During fracture healing no scar is left but a new bone tissue is created. In addition to mechanical stabilization, inflammatory cytokines, growth factors, vitamins...
EURYCOMA LONGIFOLIA, A MALAYSIAN MEDICINAL HERB, SIGNIFICANTLY UPREGULATES PROLIFERATION AND DIFFERENTIATION IN PRE-OSTEOBLASTS (MC3T3-E1): AN IN VITRO MODEL
Objective: Eurycoma longifolia (EL), a well-recognized Malaysian medicinal herb, has gained widespread popularity due to its ability to protect against bone calcium loss in androgen-deficient osteoporosis. Nevertheless...
VITRO ANTIOXIDANT PROPERTIES AND WOUND HEALING ACTIVITY OF HYDROETHANOLIC TURMERIC RHIZOME EXTRACT (ZINGIBERACEAE)
Objective: In the present study, we evaluated the ethno pharmacological effect of the different doses of Turmeric rhizome hydroethanolic extract ointment (TRO), using in vitro antioxidant activity and in vivo excision, i...
ANTIOXIDANT ACTIVITY OF FOURTEEN EULOPHIA SPECIES TRADITIONALLY KNOWN AS AMARKAND
Objective: Amarkand is a group of medicinal plants, of which tubers are regularly consumed by the tribal people in India as food and medicine. It has a strong background of regional ethnopharmacological and food uses. Th...
MITIGATING ROLE OF ZINC AND IRON AGAINST CADMIUM INDUCED TOXICITY IN LIVER AND KIDNEY OF MALE ALBINO RAT: A STUDY WITH REFERENCE TO METALLOTHIONEIN QUANTIFICATION
Objective: The present study is carried out to know the mitigating role of zinc (Zn) and / or iron (Fe) supplementation on cadmium (Cd) induced toxicity in rats with special reference to metallothionein (MT) protein.Meth...