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
Keywords
Related Articles
- EP ID EP578036
- DOI -
- Views 49
- Downloads 0
ANTIPROLIFERATIVE EFFECT OF FLOWER EXTRACTS OF SPILANTHES PANICULATA ON HEPATIC CARCINOMA CELLS
Objective: The development and evaluation of new antiproliferative drugs obtained from natural resources has gained its importance because of their less cytotoxic properties. There is no such report regarding antiprolife...
EVALUATION OF SAFETY PROFILE AND CEREBROPROTECTIVE POTENTIAL OF DIMETHYL FUMARATE (DMF) AGAINST ISCHEMIA AND REPERFUSION INDUCED CEREBRAL INJURY IN WISTAR RATS
Objective: Dimethyl fumarate (DMF) is the methyl ester of fumaric acid and used in the treatment of psoriasis, multiple sclerosis, etc. The present study was carried out to reveal safety as well as to find out the cerebr...
FREE RADICAL LOAD IN LYMPHOID ORGANS (SPLEEN AND THYMUS) OF INDIAN GOAT CAPRA HIRCUS: ROLE OF SEX, SEASON AND MELATONIN
Objective: Lymphoid organs (i.e. spleen and thymus) are important due to functional dynamicity. As a result, the generated free radicals may limit their function. Thus, present study was aimed to note seasonal and sex de...
A CASE REPORT ON ISOLATED HEMATURIA WITH ANEMIA DUE TO SCHISTOSOMA HAEMATOBIUM
An unsual case in India was reported with an Isolated Hematuria with Anemia due to Schistosoma Haematobium. A male North African patient aged with 22 years admitted in hospital in Hyderabad with chief complaints of hemat...
DESIGN OF HYDROQUINONE INDICATOR STRIP BASED ON POLYMETHYLMETHACRYLATE (PMMA) FOR IDENTIFICATION OF HYDROQUINONE IN WHITENING COSMETICS: CASE STUDY IN BANDUNG INDONESIA
Objective: The objective of this research was to developing an indicator strips made from polymer polymethylmethacrylate (PMMA) to detect hydroquinone in whitening cosmetics.Methods: The strips were prepared by reagent b...