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Evaluation of an antimicrobial L-amino acid oxidase and peptide derivatives from Bothropoides mattogrosensis pitviper venom.

Okubo BM, Silva ON, Migliolo L, Gomes DG, Porto WF, Batista CL, Ramos CS, Holanda HH, Dias SC, Franco OL, Moreno SE - PLoS ONE (2012)

Bottom Line: Otherwise, no cytotoxic activities against macrophages and erythrocytes were observed.Peptide fragments, which are the key residues involved in antimicrobial activity, were also structurally studied by using theoretical models.The fragments reported here may be promising candidates in the rational design of new antibiotics that could be used to control resistant microorganisms.

View Article: PubMed Central - PubMed

Affiliation: Programa de Pós-Graduação em Biotecnologia, Universidade Católica Dom Bosco, Campo Grande, Mato Grosso do Sul, Brazil.

ABSTRACT
Healthcare-associated infections (HAIs) are causes of mortality and morbidity worldwide. The prevalence of bacterial resistance to common antibiotics has increased in recent years, highlighting the need to develop novel alternatives for controlling these pathogens. Pitviper venoms are composed of a multifaceted mixture of peptides, proteins and inorganic components. L-amino oxidase (LAO) is a multifunctional enzyme that is able to develop different activities including antibacterial activity. In this study a novel LAO from Bothrops mattogrosensis (BmLAO) was isolated and biochemically characterized. Partial enzyme sequence showed full identity to Bothrops pauloensis LAO. Moreover, LAO here isolated showed remarkable antibacterial activity against Gram-positive and -negative bacteria, clearly suggesting a secondary protective function. Otherwise, no cytotoxic activities against macrophages and erythrocytes were observed. Finally, some LAO fragments (BmLAO-f1, BmLAO-f2 and BmLAO-f3) were synthesized and further evaluated, also showing enhanced antimicrobial activity. Peptide fragments, which are the key residues involved in antimicrobial activity, were also structurally studied by using theoretical models. The fragments reported here may be promising candidates in the rational design of new antibiotics that could be used to control resistant microorganisms.

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Related in: MedlinePlus

Multiple alignments of protein fragments determined by MALDI ToF/ToF of BmLAO against a wide diversity of L-amino oxidases from different snake species.The black coils represent α-helices and black arrows β-sheets. Numbers represent the positions of amino acid residues obtained from Gloydius halys (pdb 1tdk) L-amino-oxidase.
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pone-0033639-g002: Multiple alignments of protein fragments determined by MALDI ToF/ToF of BmLAO against a wide diversity of L-amino oxidases from different snake species.The black coils represent α-helices and black arrows β-sheets. Numbers represent the positions of amino acid residues obtained from Gloydius halys (pdb 1tdk) L-amino-oxidase.

Mentions: After de novo sequencing, the fragments obtained and selected for synthesis were denominated BmLAO-f1 (IKFEPPLPPKKAH-NH2), BmLAO-f2 (KKFWEDDG-NH2) and BmLAO-f3 (IYYPPNHNFP-NH2), respectively, in accordance with alignment (Figure 2). These three peptide fragments were chemically synthesized and further evaluated toward Gram-positive and -negative bacteria. Peptide fragments BmLAO-f1, f2 and f3 showed MIC values from 20 to 45, 125 to 250 and 110 to 220 µM for Gram-positive and –negative bacteria described in this work, and BmLAO-f1 revealed the highest activity (Table 1). In summary, natural protein BmLAO showed higher activity against microorganisms in comparison to synthetic peptides (BmLAO-f1, BmLAO-f2 and BmLAO-f3). Fragments are also interesting as biotechnological products due to the low cost incurred by their small size for synthesis facilities.


Evaluation of an antimicrobial L-amino acid oxidase and peptide derivatives from Bothropoides mattogrosensis pitviper venom.

Okubo BM, Silva ON, Migliolo L, Gomes DG, Porto WF, Batista CL, Ramos CS, Holanda HH, Dias SC, Franco OL, Moreno SE - PLoS ONE (2012)

Multiple alignments of protein fragments determined by MALDI ToF/ToF of BmLAO against a wide diversity of L-amino oxidases from different snake species.The black coils represent α-helices and black arrows β-sheets. Numbers represent the positions of amino acid residues obtained from Gloydius halys (pdb 1tdk) L-amino-oxidase.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3306279&req=5

pone-0033639-g002: Multiple alignments of protein fragments determined by MALDI ToF/ToF of BmLAO against a wide diversity of L-amino oxidases from different snake species.The black coils represent α-helices and black arrows β-sheets. Numbers represent the positions of amino acid residues obtained from Gloydius halys (pdb 1tdk) L-amino-oxidase.
Mentions: After de novo sequencing, the fragments obtained and selected for synthesis were denominated BmLAO-f1 (IKFEPPLPPKKAH-NH2), BmLAO-f2 (KKFWEDDG-NH2) and BmLAO-f3 (IYYPPNHNFP-NH2), respectively, in accordance with alignment (Figure 2). These three peptide fragments were chemically synthesized and further evaluated toward Gram-positive and -negative bacteria. Peptide fragments BmLAO-f1, f2 and f3 showed MIC values from 20 to 45, 125 to 250 and 110 to 220 µM for Gram-positive and –negative bacteria described in this work, and BmLAO-f1 revealed the highest activity (Table 1). In summary, natural protein BmLAO showed higher activity against microorganisms in comparison to synthetic peptides (BmLAO-f1, BmLAO-f2 and BmLAO-f3). Fragments are also interesting as biotechnological products due to the low cost incurred by their small size for synthesis facilities.

Bottom Line: Otherwise, no cytotoxic activities against macrophages and erythrocytes were observed.Peptide fragments, which are the key residues involved in antimicrobial activity, were also structurally studied by using theoretical models.The fragments reported here may be promising candidates in the rational design of new antibiotics that could be used to control resistant microorganisms.

View Article: PubMed Central - PubMed

Affiliation: Programa de Pós-Graduação em Biotecnologia, Universidade Católica Dom Bosco, Campo Grande, Mato Grosso do Sul, Brazil.

ABSTRACT
Healthcare-associated infections (HAIs) are causes of mortality and morbidity worldwide. The prevalence of bacterial resistance to common antibiotics has increased in recent years, highlighting the need to develop novel alternatives for controlling these pathogens. Pitviper venoms are composed of a multifaceted mixture of peptides, proteins and inorganic components. L-amino oxidase (LAO) is a multifunctional enzyme that is able to develop different activities including antibacterial activity. In this study a novel LAO from Bothrops mattogrosensis (BmLAO) was isolated and biochemically characterized. Partial enzyme sequence showed full identity to Bothrops pauloensis LAO. Moreover, LAO here isolated showed remarkable antibacterial activity against Gram-positive and -negative bacteria, clearly suggesting a secondary protective function. Otherwise, no cytotoxic activities against macrophages and erythrocytes were observed. Finally, some LAO fragments (BmLAO-f1, BmLAO-f2 and BmLAO-f3) were synthesized and further evaluated, also showing enhanced antimicrobial activity. Peptide fragments, which are the key residues involved in antimicrobial activity, were also structurally studied by using theoretical models. The fragments reported here may be promising candidates in the rational design of new antibiotics that could be used to control resistant microorganisms.

Show MeSH
Related in: MedlinePlus