Molecular cloning and characterization of the gene encoding a fibrinolytic enzyme from Bacillus subtilis Strain A1

A fibrinolytic metalloprotease gene from Bacillus subtilis has been cloned in Escheridria coliXL1-Blue and the bacterial expressed enzyme was purified. The nucleotide sequence of the cloned fibrinolytic enzyme gene revealed a single open reading frame of 1023 bp coding for 341 amino acids (M^sub r^...

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Bibliographic Details
Published in:World journal of microbiology & biotechnology 2004-10, Vol.20 (7), p.711-717
Main Authors: JEONG, Yong-Kee, JAE HYUN KIM, GAL, Sang-Wan, KIM, Ji-Eun, PARK, Soon-Suk, CHUNG, Kyung-Tae, KIM, Young-Hee, KIM, Byung-Woo, JOO, Woo-Hong
Format: Article
Language:English
Subjects:
Amino acids
Bacillus anthracis
Bacillus subtilis
Bacteria
Bacteriology
Biological and medical sciences
Biotechnology
Cloning
E coli
Enzymes
Fundamental and applied biological sciences. Psychology
Microbiology
Serratia marcescens
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Summary:A fibrinolytic metalloprotease gene from Bacillus subtilis has been cloned in Escheridria coliXL1-Blue and the bacterial expressed enzyme was purified. The nucleotide sequence of the cloned fibrinolytic enzyme gene revealed a single open reading frame of 1023 bp coding for 341 amino acids (M^sub r^ 37708.21 Da). N-terminal amino acid sequencing of the fibrinolytic enzyme excreted from E. coli host cells revealed that the mature fibrinolytic enzyme consists of 288 amino acids (M^sub r^ 31391.1 Da). The deduced amino acid sequence showed significant homology with Erwina carotovora neutral metalloprotease and Serratia marcescens minor metalloprotease by 65 and 58% amino acid sequence identity, respectively. The protein showed significant alignments with the conserved domain of catalytic activity and the α-helix domain in Bacillus anthracisthermolysis metalloprotease. The biochemical properties of the purified enzyme suggested that the enzyme is a fibrinolytic metalloprotease, which has optimal activity at pH 7.0 and 50 °C.[PUBLICATION ABSTRACT]
ISSN:0959-3993
1573-0972
DOI:10.1007/s11274-003-4514-5