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Toxicon : official journal of the International Society on Toxinology2008; 53(3); 330-341; doi: 10.1016/j.toxicon.2008.12.004

Antigenic, microbicidal and antiparasitic properties of an l-amino acid oxidase isolated from Bothrops jararaca snake venom.

Abstract: Venoms from the bee Apis mellifera, the caterpillar Lonomia achelous, the spiders Lycosa sp. and Phoneutria nigriventer, the scorpions Tityus bahiensis and Tityus serrulatus, and the snakes Bothrops alternatus, Bothrops jararaca, Bothrops jararacussu, Bothrops moojeni, Bothrops neuwiedi, Crotalus durissus terrificus, and Lachesis muta were assayed (800mug/mL) for activity against Staphylococcus aureus. Venoms from B. jararaca and B. jararacussu showed the highest S. aureus growth inhibition and also against other Gram-positive and Gram-negative bacteria. To characterize the microbicidal component(s) produced by B. jararaca, venom was fractionated through gel exclusion chromatography. The high molecular weight, anti-S. aureus P1 fraction was further resolved by anion exchange chromatography through Mono Q columns using a 0-0.5M NaCl gradient. Bactericidal Mono Q fractions P5 and P6 showed significant LAAO activity using l-leucine as substrate. These fractions were pooled and subjected to Heparin affinity chromatography, which rendered a single LAAO activity peak. The anti-S. aureus activity was abolished by catalase, suggesting that the effect is dependent on H(2)O(2) production. SDS-PAGE of isolated LAAO indicated the presence of three isoforms since deglycosylation with a recombinant N-glycanase rendered a single 38.2 kDa component. B. jararaca LAAO specific activity was 142.7 U/mg, based on the oxidation of l-leucine. The correlation between in vivo neutralization of lethal toxicity (ED(50)) and levels of horse therapeutic antibodies anti-LAAO measured by ELISA was investigated to predict the potency of Brazilian antibothropic antivenoms. Six horses were hyperimmunized with Bothrops venoms (50% from B. jararaca and 12.5% each from B. alternatus, B. jararacussu, B. neuwiedii and B. moojeni). To set up an indirect ELISA, B. jararaca LAAO and crude venom were used as antigens. Correlation coefficients (r) between ED(50) and ELISA antibody titers against B. jararaca venom and LAAO were 0.846 (p<0.001) and 0.747 (p<0.001), respectively. The hemolytic and leishmanicidal (anti-Leishmania amazonensis) activity of LAAO was also determined.
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Publication Date: 2008-12-11 PubMed ID: 19101583DOI: 10.1016/j.toxicon.2008.12.004Google Scholar: Lookup
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  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This study evaluates the potential of the venom from Bothrops jararaca, a type of snake, as a source of antiparasitic, antibacterial, and antigenic agents. Extracts from the venom showed high efficacy in inhibiting the growth of Staphylococcus aureus, as well as other Gram-positive and Gram-negative bacteria, an activity linked to the enzymatic action of l-amino acid oxidase (LAAO) found in the venom.

Research Procedure

  • Venom samples were taken from various organisms like bees, spiders, scorpions, caterpillars, and snakes.
  • These samples were tested for their inhibitory effects on the bacterial strain Staphylococcus aureus.
  • The venoms of B. jararaca and B. jararacussu snakes demonstrated the highest inhibition of S. aureus growth.
  • Further analysis was performed on B. jararaca venom, aiming to isolate and identify the microbicidal components. To this end, the venom was fractionated and passed through various steps of chromatography.
  • Fractions of the venom that showed bactericidal effects also had significant LAAO activity.
  • The active component was identified to depend on hydrogen peroxide (H2O2) production, as its antimicrobial effect was eliminated by catalase, an enzyme that breaks down hydrogen peroxide.
  • LAAO from the venom was found to have three isoforms, which were reduced to a single component after deglycosylation, a process that removes sugar groups attached to the protein.
  • The specific activity of LAAO was quantitatively defined based on the oxidation of the amino acid l-leucine.

Antivenom Development

  • Tests were carried out to find a correlation between the levels of anti-LAAO antibodies and the neutralization of venom’s lethal toxicity.
  • Six horses were immunized with Bothrops venoms, and the potency of the derived antivenom was measured using an indirect ELISA (Enzyme-Linked Immunosorbent Assay).
  • A high correlation was found between the potency of antivenom (determined by the capacity to neutralize venom toxicity) and the levels of antibodies against both the total venom and LAAO.
  • This suggests the potential efficacy of these antibodies in neutralizing venom toxicity, presenting an avenue for antivenom development.

Other Properties

  • The venoms were also analyzed for hemolytic activity (ability to break down red blood cells) and leishmanicidal activity, showing potential effects against the parasite Leishmania amazonensis responsible for causing Leishmaniasis, a tropical disease.

Cite This Article

APA
Ciscotto P, Machado de Avila RA, Coelho EA, Oliveira J, Diniz CG, Farías LM, de Carvalho MA, Maria WS, Sanchez EF, Borges A, Chávez-Olórtegui C. (2008). Antigenic, microbicidal and antiparasitic properties of an l-amino acid oxidase isolated from Bothrops jararaca snake venom. Toxicon, 53(3), 330-341. https://doi.org/10.1016/j.toxicon.2008.12.004

Publication

Toxicon : official journal of the International Society on Toxinology
ISSN: 0041-0101
NlmUniqueID: 1307333
Country: England
Language: English
Volume: 53
Issue: 3
Pages: 330-341

Researcher Affiliations

Ciscotto, P
  • Departamento de Bioquímica e Imunologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Av. Antonio Carlos 6627, 30161-970 Belo Horizonte, Minas Gerais, Brazil.
Machado de Avila, R A
    Coelho, E A F
      Oliveira, J
        Diniz, C G
          Farías, L M
            de Carvalho, M A R
              Maria, W S
                Sanchez, E F
                  Borges, A
                    Chávez-Olórtegui, C

                      MeSH Terms

                      • Animals
                      • Antibodies / blood
                      • Biological Assay
                      • Bothrops
                      • Chromatography, Affinity
                      • Chromatography, Gel
                      • Chromatography, Ion Exchange
                      • Electrophoresis, Polyacrylamide Gel
                      • Enzyme-Linked Immunosorbent Assay
                      • Horses
                      • L-Amino Acid Oxidase / immunology
                      • L-Amino Acid Oxidase / isolation & purification
                      • L-Amino Acid Oxidase / pharmacology
                      • Lethal Dose 50
                      • Neutralization Tests
                      • Staphylococcus aureus / drug effects
                      • Viper Venoms / enzymology
                      • Viper Venoms / toxicity

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