Isolation and structural analysis of bamylocin A, novel lipopeptide from Bacillus amyloliquefaciens LP03 having antagonistic and crude oil-emulsifying activity.
Bacillus amyloliquefaciens strain LP03 isolated from soil, produced an antagonistic compound that strongly inhibited the growth of plant-pathogenic fungi and a lipopeptide biosurfactant. Also, isolated strain LP03 had a marked crude oil-emulsifying activity as it developed a clear zone around the colony after incubation for 24 h at 37 degrees C. LP03 was identified as Bacillus amyloliquefaciens by analysis of partial 16 S rRNA gene and partial gyrA gene sequence. The lipopeptide was purified by acid precipitation of cell-free culture broth, extraction of the precipitates with methanol, silica gel column chromatography, and reverse-phase, high-pressure liquid chromatography. The purified biosurfactant was analyzed biochemical structure by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) and electrospray ionization mass spectrometry/mass spectrometry (ESI-MS/MS). The masses of the two peaks were observed by HPLC chromatography. Their masses were determined to be 1,044 and 1,058 m/z with MALDI-TOF mass spectrometry. As constituents of the peptide and lipophilic part of the m/z 1,022.6, seven amino acids (Glu-Leu-Met-Leu-Pro-Leu-Leu) and beta-hydroxy-C13 fatty acid were determined by ESI-MS/MS. The lipopeptide of 1,022.6 Da differed from surfactins in the substitution of leucine, valine and aspartic acid in positions 3, 4, and 5 by methionine, leucine, and proline, respectively. Novel lipopeptide was designated as bamylocin A.
Lee
Sang-Cheol
SC
Department of Biotechnology, Faculty of Natural Resources and Life Science, Dong-A University, Busan 604-714, Korea.
Kim
Sun-Hee
SH
Park
In-Hye
IH
Chung
Soo-Yeol
SY
Choi
Yong-Lark
YL
D000935:Antifungal Agents / Q000302:isolation & purification* / Q000494:pharmacology*; D001407:Bacillus / Q000737:chemistry* / Q000145:classification / Q000302:isolation & purification; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D027081:DNA Gyrase / Q000235:genetics; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D045703:Emulsifying Agents / Q000302:isolation & purification* / Q000494:pharmacology*; D005227:Fatty Acids / Q000032:analysis; D005658:Fungi / Q000187:drug effects; D007723:Korea; D008074:Lipoproteins / Q000302:isolation & purification* / Q000494:pharmacology*; D008970:Molecular Weight; D010578:Petroleum / Q000378:metabolism; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D020539:Sequence Analysis, Protein; D012988:Soil Microbiology; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D000935:Antifungal Agents; D004271:DNA, Fungal; D004275:DNA, Ribosomal; D045703:Emulsifying Agents; D005227:Fatty Acids; D008074:Lipoproteins; D010578:Petroleum; D012336:RNA, Ribosomal, 16S; D027081:DNA Gyrase