Open reading frame yjbI of Bacillus subtilis codes for truncated hemoglobin.
A hypothetical open reading frame from Bacillus subtilis genome, yjbI [NCBI genome database Accession No. ] having homology to many globin and globin-like proteins from different microbial genomes, was selectively amplified from the chromosomal DNA of B. subtilis strain DB104 based on genome sequence database of B. subtilis strain 168. The gene was cloned and over-expressed in Escherichia coli under the transcriptional control of tandem lambda P(L) and P(R) promoters, and the protein was purified to homogeneity. The single-chain monomeric hemoglobin-like protein is stable to the extent of 5.45 kcal/mol at 25 degrees C, binds carbon mono-oxide, and shows optical spectra characteristic of hemoproteins. The protein also exhibits peroxidase-like activity. This is the first report of a truncated bacterial globin endowed with peroxidase-like activity. The activity is enhanced in the presence of urea and guanidine hydrochloride, more so in the presence of the latter. Presumably, only a small portion of the protein is involved in peroxidase activity, which is exposed with increasing concentration of the denaturants.
Choudhary
Manohar L
ML
Bharat Biotech Foundation, Genome Valley, Shameerpet, Hyderabad 500 078, India.
Jawaid
Safdar
S
Ahuja
Munish K
MK
Shiva
Narayana K
NK
Gupta
Pankaj
P
Bhuyan
Abani K
AK
Khatri
Ghan Shyam
GS
D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000737:chemistry / Q000235:genetics*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D003001:Cloning, Molecular; D004795:Enzyme Stability; D004926:Escherichia coli / Q000235:genetics; D015964:Gene Expression Regulation, Bacterial; D006454:Hemoglobins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010544:Peroxidases / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D010802:Phylogeny; D011487:Protein Conformation; D017510:Protein Folding; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D054793:Truncated Hemoglobins
D001426:Bacterial Proteins; D006454:Hemoglobins; D054793:Truncated Hemoglobins; D010544:Peroxidases