Two Bacillus beta-mannanases having different COOH termini are produced in Escherichia coli carrying pMAH5.
The nucleotide sequence was determined for the alkalophilic Bacillus sp. strain AM-001 beta-mannanase gene which produced two beta-mannanases (A and B) in Escherichia coli transformants. The putative beta-mannanase gene was 1,539 base pairs long and encoded a mature beta-mannanase protein of 487 amino acids and a signal peptide of 26 amino acids. The COOH-terminal amino acid of beta-mannanase A is an arginine residue located at amino acid 513 of the deduced amino acid sequence, and that of beta-mannanase B is a valine residue located at amino acid 365. Deletion derivatives having 1,098 base pairs from the ATG start codon maintained the beta-mannanase activity of the encoded polypeptide. However, clones harboring DNA fragments (1,051 base pairs) shorter than the gene which encoded beta-mannanase B (1,095 base pairs) did not exhibit the beta-mannanase activity. The simultaneous production of both beta-mannanases A and B in an E. coli transformant was demonstrated by the maxicell procedure.
Akino
T
T
Laboratory of Microbial Metabolism, Research Development Corporation of Japan, Tokyo.
Kato
C
C
Horikoshi
K
K
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D001345:Autoradiography; D001407:Bacillus / Q000201:enzymology / Q000235:genetics*; D001426:Bacterial Proteins / Q000032:analysis; D001483:Base Sequence; D015153:Blotting, Western; D003062:Codon / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D008361:Mannosidases / Q000096:biosynthesis / Q000235:genetics*; D008969:Molecular Sequence Data; D010957:Plasmids; D044902:beta-Mannosidase
D000596:Amino Acids; D001426:Bacterial Proteins; D003062:Codon; D004269:DNA, Bacterial; D008361:Mannosidases; D044902:beta-Mannosidase