title: Production of recombinant streptokinase from Streptococcus pyogenes isolate and its potential for thrombolytic therapy.
abstract: OBJECTIVE
  To produce an effective recombinant streptokinase (rSK) from pathogenic Streptococcus pyogenes isolate in yeast, and evaluate its potential for thrombolytic therapy.
  
  
  METHODS
  This study was conducted from November 2012 to December 2013 at King Khalid University, Abha, Kingdom of Saudi Arabia (KSA). Throat swabs collected from 45 pharyngitis patients in Asser Central Hospital, Abha, KSA were used to isolate Streptococcus pyogenes. The bacterial DNA was used for amplification of the streptokinase gene (1200 bp). The gene was cloned and in vitro transcribed in an eukaryotic expression vector that was transformed into yeast Pichia pastoris SMD1168, and the rSK protein was purified and tested for its thrombolytic activity.
  
  
  RESULTS
  The Streptococcus pyogenes strain was isolated and its DNA nucleotide sequence revealed similarity to other Streptococcus pyogenes in the Gene bank. Sequencing of the amplified gene based on DNA nucleotide sequence revealed a SK gene closely related to other SK genes in the Gene bank. However, based on deduced amino acids sequence, the gene formed a separate cluster different from clusters formed by other examined genes, suggesting a new bacterial isolate and accordingly a new gene. The purified protein showed 82% clot lysis compared to a commercial SK (81%) at an enzyme concentration of 2000 U/ml.
  
  
  CONCLUSIONS
  The present yeast rSK showed similar thrombolytic activity in vitro as that of a commercial SK, suggesting its potential for thrombolytic therapy and large scale production.
authors.affiliation: Department of Internal Medicine, College of Medicine, King Khalid University, Abha, Kingdom of Saudi Arabia. E-mail. aassiri@yahoo.com.
authors.forename: Abdullah S
authors.initials: AS
authors.lastname: Assiri
authors.forename: Basiouny A
authors.initials: BA
authors.lastname: El-Gamal
authors.forename: Elsayed E
authors.initials: EE
authors.lastname: Hafez
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Haidara
mesh_terms: D063285:Fibrin Clot Lysis Time; D005343:Fibrinolytic Agents / Q000494:pharmacology*; D005822:Genetic Vectors; D006801:Humans; D010612:Pharyngitis / Q000382:microbiology; D010843:Pichia; D011994:Recombinant Proteins / Q000096:biosynthesis; D013290:Streptococcal Infections / Q000382:microbiology; D013297:Streptococcus pyogenes / Q000235:genetics* / Q000302:isolation & purification; D013300:Streptokinase / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D013917:Thrombin / Q000187:drug effects; D015912:Thrombolytic Therapy
chemical_list: D005343:Fibrinolytic Agents; D011994:Recombinant Proteins; D013300:Streptokinase; D013917:Thrombin