Mushtaq, Z.
Isolation, identification and sequence analysis of subtilisin gene (quaking homolog) encoding a fibrinolytic enzyme from bacillus subtilis - Vol.83(4), July-Aug - Mumbai Indian Journal of Pharmaceutical Science 2021 - 856-864p.
Fibrinolytic therapy progressed by the evolution of different strategies that helped in enhancing its efficacy
and specificity. The use of microbial fibrinolytic enzymes is leading to a promising direction for the treatment
of cardiovascular diseases. Subtilisin, a member of subtilases is a fibrinolytic enzyme abundantly found in
Bacillus species. The isolation of subtilisin gene (quaking homolog) from Bacillus subtilis was attempted in
the present work. The genomic deoxyribonucleic acid extraction was done following Yamada protocol and
used as a template for polymerase chain reaction amplification of the target gene using specifically designed
primers. The polymerase chain reaction product was ligated into cloning vector pTZ57R/T followed by its
transformation into Escherichia coli top 10 strain. A 1090 base pair, partial gene sequence was amplified
coding for subtilisin protein of 363 amino acids with molecular weight of 37550.7 Daltons. The nucleotide
sequence revealed significant evolutionary relationships with subtilases from other strains of Bacillus
subtilis. Our study confirms the presence of subtilisin (quaking homolog) gene in local Bacillus species
suggesting economical way to produce important thrombolytic agents of commercial and pharmaceutical
worth.
PHARMACEUTICS
Isolation, identification and sequence analysis of subtilisin gene (quaking homolog) encoding a fibrinolytic enzyme from bacillus subtilis - Vol.83(4), July-Aug - Mumbai Indian Journal of Pharmaceutical Science 2021 - 856-864p.
Fibrinolytic therapy progressed by the evolution of different strategies that helped in enhancing its efficacy
and specificity. The use of microbial fibrinolytic enzymes is leading to a promising direction for the treatment
of cardiovascular diseases. Subtilisin, a member of subtilases is a fibrinolytic enzyme abundantly found in
Bacillus species. The isolation of subtilisin gene (quaking homolog) from Bacillus subtilis was attempted in
the present work. The genomic deoxyribonucleic acid extraction was done following Yamada protocol and
used as a template for polymerase chain reaction amplification of the target gene using specifically designed
primers. The polymerase chain reaction product was ligated into cloning vector pTZ57R/T followed by its
transformation into Escherichia coli top 10 strain. A 1090 base pair, partial gene sequence was amplified
coding for subtilisin protein of 363 amino acids with molecular weight of 37550.7 Daltons. The nucleotide
sequence revealed significant evolutionary relationships with subtilases from other strains of Bacillus
subtilis. Our study confirms the presence of subtilisin (quaking homolog) gene in local Bacillus species
suggesting economical way to produce important thrombolytic agents of commercial and pharmaceutical
worth.
PHARMACEUTICS