Biosynthesis and transcriptional analysis of thurincin H, a tandem repeated bacteriocin genetic locus, produced by Bacillus thuringiensis SF361.
Thurincin H, a bacteriocin produced by Bacillus thuringiensis SF361 isolated from honey, strongly inhibited the growth of Bacillus cereus F4552. The bacteriocin was purified by 65% ammonium sulfate precipitation of the culture supernatant, followed by octyl-sepharose CL-4B and reverse-phase HPLC. The molecular mass of the bacteriocin was determined to be 3139.51 Da and the 14 amino acids of the bacteriocin at the N-terminus were identified. The complete amino acid sequence of mature thurincin H was deduced from three structural genes, thnA1, thnA2, and thnA3 found in tandem repeats on the chromosome, all of which encode for the same bacteriocin, thurincin H. The genetic determinants for thurincin H biosynthesis consist of 10 ORFs, including three thurincin H structural genes. Northern hybridization elucidated that the transcription of all three bacteriocin structural genes was regulated by a putative promoter located upstream of thnA1.
Lee
Hyungjae
H
Department of Food Science and Technology, New York State Agricultural Experiment Station, Cornell University, 630 W North St., Geneva, NY 14456, USA.
Churey
John J
JJ
Worobo
Randy W
RW
D000595:Amino Acid Sequence; D000645:Ammonium Sulfate / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000096:biosynthesis* / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D001409:Bacillus cereus / Q000187:drug effects; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D001483:Base Sequence; D053898:Biosynthetic Pathways / Q000235:genetics; D005591:Chemical Fractionation / Q000379:methods; D002853:Chromatography, Liquid / Q000379:methods; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D020440:Gene Duplication; D020869:Gene Expression Profiling; D023061:Gene Order; D006722:Honey / Q000382:microbiology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011401:Promoter Regions, Genetic; D012685:Sepharose; D017422:Sequence Analysis, DNA
D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D004269:DNA, Bacterial; D012685:Sepharose; D000645:Ammonium Sulfate