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Isolation and characterization of marine Brevibacillus sp. S-1 collected from South China Sea and a novel antitumor peptide produced by the strain.

Zheng L, Yi Y, Liu J, Lin X, Yang K, Lv M, Zhou X, Hao J, Liu J, Zheng Y, Sun M - PLoS ONE (2014)

Bottom Line: S-1, using ion-exchange chromatography and reverse-phase HPLC chromatography.Additionally, SBP exhibited low cytotoxicity against HFL1 human normal fibroblast lung cells.The result suggested that the cytotoxic effect of the peptide is specific to tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, PR China.

ABSTRACT
A Gram-positive, rod-shaped bacterium, designated as S-1, was isolated from a marine sediment sample collected from South China Sea. Phylogenetic analysis based on 16S rRNA gene sequence showed that S-1 belongs to the genus Brevibacillus. A novel cytotoxic peptide was isolated from the fermentation broth of the marine-derived bacterium Brevibacillus sp. S-1, using ion-exchange chromatography and reverse-phase HPLC chromatography. The molecular weight of this peptide was determined as 1570 Da by MALDI-TOF mass spectrometry, and its structure was proposed as a cyclic peptide elucidated by MALDI-TOF/TOF mass spectrometry and de novo sequencing. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay showed that this peptide exhibited cytotoxicity against BEL-7402 human hepatocellular carcinoma cells, RKO human colon carcinoma cells, A549 human lung carcinoma cells, U251 human glioma cells and MCF-7 human breast carcinoma cells. Additionally, SBP exhibited low cytotoxicity against HFL1 human normal fibroblast lung cells. The result suggested that the cytotoxic effect of the peptide is specific to tumor cells.

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Related in: MedlinePlus

The molecular ion peak of SBP [M+H]+1 to 1570.0358 measured by MALDI TOF/TOF Mass spectrum.
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pone-0111270-g004: The molecular ion peak of SBP [M+H]+1 to 1570.0358 measured by MALDI TOF/TOF Mass spectrum.

Mentions: Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI TOF/TOF MS) analysis of SBP was performed with different ion signals in the mass range of 700–3600 Da. 1570.0358, 1592.0237 and 1608.2003 is M+H, M+Na and M+K ions, respectively. The molecular weight of SBP was about 1570 Da, as determined by MALDI TOF/TOF MS analysis (Figure 4). The precursor ions (m/z 1570. 0358) were further detected by MS/MS analysis. MS/MS spectra consisting of a series of y and b ions were obtained and used for de novo sequencing. Based on the manual calculation of the molecular weights and the m/z values, the amino acid sequence of SBP peptide was proposed. The amino acid sequence of fragment ion m/z 1570.0358 was the APQNI/LVPKTI/LKYI/LC and the peptide was structurally cycle (Figure 5). One of the amino acids in this sequence is Thr, and therefore, SBP may be a cyclic peptide linked end-to-end, or a cyclic depsipeptide linked with a Thr-(-OH)-to-Cys(-COOH) bond. However, the possibility of being a cyclic desipeptide is quite low because of bigger space steric hindrance between Thr and Cys. The theoretical M+H ion of the proposed cyclic peptide is 1569.925 Da, and in agreement with the exact mass observed for the M+H ion (figure 4). The molecular weight of Ile and Leu is same, so these two amino acids could not be determined according to available data. The results of the present study indicated that the amino acid sequence of SBP was the APQNI/LVPKTI/LKYI/LC, and the SBP was most likely to be a cyclic peptide linked in an end-to-end fashion. The amino acid sequence alignment was performed against the National Center for Biotechnology Information Basic Local Alignment Search Tool (NCBI BLAST) database, and the low similarity of its amino acid sequences with the known proteins indicated that SBP was a novel peptide.


Isolation and characterization of marine Brevibacillus sp. S-1 collected from South China Sea and a novel antitumor peptide produced by the strain.

Zheng L, Yi Y, Liu J, Lin X, Yang K, Lv M, Zhou X, Hao J, Liu J, Zheng Y, Sun M - PLoS ONE (2014)

The molecular ion peak of SBP [M+H]+1 to 1570.0358 measured by MALDI TOF/TOF Mass spectrum.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4220994&req=5

pone-0111270-g004: The molecular ion peak of SBP [M+H]+1 to 1570.0358 measured by MALDI TOF/TOF Mass spectrum.
Mentions: Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI TOF/TOF MS) analysis of SBP was performed with different ion signals in the mass range of 700–3600 Da. 1570.0358, 1592.0237 and 1608.2003 is M+H, M+Na and M+K ions, respectively. The molecular weight of SBP was about 1570 Da, as determined by MALDI TOF/TOF MS analysis (Figure 4). The precursor ions (m/z 1570. 0358) were further detected by MS/MS analysis. MS/MS spectra consisting of a series of y and b ions were obtained and used for de novo sequencing. Based on the manual calculation of the molecular weights and the m/z values, the amino acid sequence of SBP peptide was proposed. The amino acid sequence of fragment ion m/z 1570.0358 was the APQNI/LVPKTI/LKYI/LC and the peptide was structurally cycle (Figure 5). One of the amino acids in this sequence is Thr, and therefore, SBP may be a cyclic peptide linked end-to-end, or a cyclic depsipeptide linked with a Thr-(-OH)-to-Cys(-COOH) bond. However, the possibility of being a cyclic desipeptide is quite low because of bigger space steric hindrance between Thr and Cys. The theoretical M+H ion of the proposed cyclic peptide is 1569.925 Da, and in agreement with the exact mass observed for the M+H ion (figure 4). The molecular weight of Ile and Leu is same, so these two amino acids could not be determined according to available data. The results of the present study indicated that the amino acid sequence of SBP was the APQNI/LVPKTI/LKYI/LC, and the SBP was most likely to be a cyclic peptide linked in an end-to-end fashion. The amino acid sequence alignment was performed against the National Center for Biotechnology Information Basic Local Alignment Search Tool (NCBI BLAST) database, and the low similarity of its amino acid sequences with the known proteins indicated that SBP was a novel peptide.

Bottom Line: S-1, using ion-exchange chromatography and reverse-phase HPLC chromatography.Additionally, SBP exhibited low cytotoxicity against HFL1 human normal fibroblast lung cells.The result suggested that the cytotoxic effect of the peptide is specific to tumor cells.

View Article: PubMed Central - PubMed

Affiliation: Key Laboratory of Sustainable Development of Marine Fisheries, Ministry of Agriculture, Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao, PR China.

ABSTRACT
A Gram-positive, rod-shaped bacterium, designated as S-1, was isolated from a marine sediment sample collected from South China Sea. Phylogenetic analysis based on 16S rRNA gene sequence showed that S-1 belongs to the genus Brevibacillus. A novel cytotoxic peptide was isolated from the fermentation broth of the marine-derived bacterium Brevibacillus sp. S-1, using ion-exchange chromatography and reverse-phase HPLC chromatography. The molecular weight of this peptide was determined as 1570 Da by MALDI-TOF mass spectrometry, and its structure was proposed as a cyclic peptide elucidated by MALDI-TOF/TOF mass spectrometry and de novo sequencing. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay showed that this peptide exhibited cytotoxicity against BEL-7402 human hepatocellular carcinoma cells, RKO human colon carcinoma cells, A549 human lung carcinoma cells, U251 human glioma cells and MCF-7 human breast carcinoma cells. Additionally, SBP exhibited low cytotoxicity against HFL1 human normal fibroblast lung cells. The result suggested that the cytotoxic effect of the peptide is specific to tumor cells.

Show MeSH
Related in: MedlinePlus