A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity

physicochemical characterization

Angelo M L Denadai, Marcelo M Santoro, Miriam T P Lopes, Angélica Chenna, Frederico B de Sousa, Gabriela Mol Avelar, Marco R Túlio Gomes, Fanny Guzman, Carlos E Salas, Rubén D Sinisterra

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

BACKGROUND: Cyclodextrins are suitable drug delivery systems because of their ability to subtly modify the physical, chemical, and biological properties of guest molecules through labile interactions by formation of inclusion and/or association complexes. Plant cysteine proteinases from Caricaceae and Bromeliaceae are the subject of therapeutic interest, because of their anti-inflammatory, antitumoral, immunogenic, and wound-healing properties.

METHODS: In this study, we analyzed the association between beta-cyclodextrin (betaCD) and fraction P1G10 containing the bioactive proteinases from Carica candamarcensis, and described the physicochemical nature of the solid-state self-assembled complexes by Fourier transform infrared (FTIR) spectroscopy, thermogravimetry (TG), differential scanning calorimetry (DSC), X-ray powder diffraction (XRD), and nuclear magnetic resonance (NMR), as well as in solution by circular dichroism (CD), isothermal titration calorimetry (ITC), and amidase activity.

RESULTS AND DISCUSSION: The physicochemical analyses suggest the formation of a complex between P1G10 and betaCD. Higher secondary interactions, namely hydrophobic interactions, hydrogen bonding and van der Waals forces were observed at higher P1G10 : betaCD mass ratios. These results provide evidence of the occurrence of strong solid-state supramolecular non-covalent interactions between P1G10 and betaCD. Microcalorimetric analysis demonstrates that complexation results in a favorable enthalpic contribution, as has already been described during formation of similar betaCD inclusion compounds. The amidase activity of the complex shows that the enzyme activity is not readily available at 24 hours after dissolution of the complex in aqueous buffer; the proteinase becomes biologically active by the second day and remains stable until day 16, when a gradual decrease occurs, with basal activity attained by day 29.

CONCLUSION: The reported results underscore the potential for betaCDs as candidates for complexing cysteine proteinases, resulting in supramolecular arrays with sustained proteolytic activity.

Original languageEnglish
Pages (from-to)283-91
Number of pages9
JournalBioDrugs
Volume20
Issue number5
Publication statusPublished - 2006

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amidase
Peptide Hydrolases
Cysteine Proteases
Caricaceae
Bromeliaceae
Thermogravimetry
Carica
Powder Diffraction
Calorimetry
Differential Scanning Calorimetry
Cyclodextrins
Fourier Transform Infrared Spectroscopy
Hydrogen Bonding
Drug Delivery Systems
Circular Dichroism
Hydrophobic and Hydrophilic Interactions
X-Ray Diffraction
Wound Healing
Buffers
Anti-Inflammatory Agents

Keywords

  • Amidohydrolases
  • Calorimetry
  • Calorimetry, Differential Scanning
  • Carica
  • Circular Dichroism
  • Latex
  • Magnetic Resonance Spectroscopy
  • Peptide Hydrolases
  • Spectroscopy, Fourier Transform Infrared
  • Thermogravimetry
  • X-Ray Diffraction
  • beta-Cyclodextrins

Cite this

Denadai, A. M. L., Santoro, M. M., Lopes, M. T. P., Chenna, A., de Sousa, F. B., Mol Avelar, G., ... Sinisterra, R. D. (2006). A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity: physicochemical characterization. BioDrugs, 20(5), 283-91.

A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity : physicochemical characterization. / Denadai, Angelo M L; Santoro, Marcelo M; Lopes, Miriam T P; Chenna, Angélica; de Sousa, Frederico B; Mol Avelar, Gabriela; Gomes, Marco R Túlio; Guzman, Fanny; Salas, Carlos E; Sinisterra, Rubén D.

In: BioDrugs, Vol. 20, No. 5, 2006, p. 283-91.

Research output: Contribution to journalArticle

Denadai, AML, Santoro, MM, Lopes, MTP, Chenna, A, de Sousa, FB, Mol Avelar, G, Gomes, MRT, Guzman, F, Salas, CE & Sinisterra, RD 2006, 'A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity: physicochemical characterization', BioDrugs, vol. 20, no. 5, pp. 283-91.
Denadai AML, Santoro MM, Lopes MTP, Chenna A, de Sousa FB, Mol Avelar G et al. A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity: physicochemical characterization. BioDrugs. 2006;20(5):283-91.
Denadai, Angelo M L ; Santoro, Marcelo M ; Lopes, Miriam T P ; Chenna, Angélica ; de Sousa, Frederico B ; Mol Avelar, Gabriela ; Gomes, Marco R Túlio ; Guzman, Fanny ; Salas, Carlos E ; Sinisterra, Rubén D. / A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity : physicochemical characterization. In: BioDrugs. 2006 ; Vol. 20, No. 5. pp. 283-91.
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T1 - A supramolecular complex between proteinases and beta-cyclodextrin that preserves enzymatic activity

T2 - physicochemical characterization

AU - Denadai, Angelo M L

AU - Santoro, Marcelo M

AU - Lopes, Miriam T P

AU - Chenna, Angélica

AU - de Sousa, Frederico B

AU - Mol Avelar, Gabriela

AU - Gomes, Marco R Túlio

AU - Guzman, Fanny

AU - Salas, Carlos E

AU - Sinisterra, Rubén D

PY - 2006

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N2 - BACKGROUND: Cyclodextrins are suitable drug delivery systems because of their ability to subtly modify the physical, chemical, and biological properties of guest molecules through labile interactions by formation of inclusion and/or association complexes. Plant cysteine proteinases from Caricaceae and Bromeliaceae are the subject of therapeutic interest, because of their anti-inflammatory, antitumoral, immunogenic, and wound-healing properties.METHODS: In this study, we analyzed the association between beta-cyclodextrin (betaCD) and fraction P1G10 containing the bioactive proteinases from Carica candamarcensis, and described the physicochemical nature of the solid-state self-assembled complexes by Fourier transform infrared (FTIR) spectroscopy, thermogravimetry (TG), differential scanning calorimetry (DSC), X-ray powder diffraction (XRD), and nuclear magnetic resonance (NMR), as well as in solution by circular dichroism (CD), isothermal titration calorimetry (ITC), and amidase activity.RESULTS AND DISCUSSION: The physicochemical analyses suggest the formation of a complex between P1G10 and betaCD. Higher secondary interactions, namely hydrophobic interactions, hydrogen bonding and van der Waals forces were observed at higher P1G10 : betaCD mass ratios. These results provide evidence of the occurrence of strong solid-state supramolecular non-covalent interactions between P1G10 and betaCD. Microcalorimetric analysis demonstrates that complexation results in a favorable enthalpic contribution, as has already been described during formation of similar betaCD inclusion compounds. The amidase activity of the complex shows that the enzyme activity is not readily available at 24 hours after dissolution of the complex in aqueous buffer; the proteinase becomes biologically active by the second day and remains stable until day 16, when a gradual decrease occurs, with basal activity attained by day 29.CONCLUSION: The reported results underscore the potential for betaCDs as candidates for complexing cysteine proteinases, resulting in supramolecular arrays with sustained proteolytic activity.

AB - BACKGROUND: Cyclodextrins are suitable drug delivery systems because of their ability to subtly modify the physical, chemical, and biological properties of guest molecules through labile interactions by formation of inclusion and/or association complexes. Plant cysteine proteinases from Caricaceae and Bromeliaceae are the subject of therapeutic interest, because of their anti-inflammatory, antitumoral, immunogenic, and wound-healing properties.METHODS: In this study, we analyzed the association between beta-cyclodextrin (betaCD) and fraction P1G10 containing the bioactive proteinases from Carica candamarcensis, and described the physicochemical nature of the solid-state self-assembled complexes by Fourier transform infrared (FTIR) spectroscopy, thermogravimetry (TG), differential scanning calorimetry (DSC), X-ray powder diffraction (XRD), and nuclear magnetic resonance (NMR), as well as in solution by circular dichroism (CD), isothermal titration calorimetry (ITC), and amidase activity.RESULTS AND DISCUSSION: The physicochemical analyses suggest the formation of a complex between P1G10 and betaCD. Higher secondary interactions, namely hydrophobic interactions, hydrogen bonding and van der Waals forces were observed at higher P1G10 : betaCD mass ratios. These results provide evidence of the occurrence of strong solid-state supramolecular non-covalent interactions between P1G10 and betaCD. Microcalorimetric analysis demonstrates that complexation results in a favorable enthalpic contribution, as has already been described during formation of similar betaCD inclusion compounds. The amidase activity of the complex shows that the enzyme activity is not readily available at 24 hours after dissolution of the complex in aqueous buffer; the proteinase becomes biologically active by the second day and remains stable until day 16, when a gradual decrease occurs, with basal activity attained by day 29.CONCLUSION: The reported results underscore the potential for betaCDs as candidates for complexing cysteine proteinases, resulting in supramolecular arrays with sustained proteolytic activity.

KW - Amidohydrolases

KW - Calorimetry

KW - Calorimetry, Differential Scanning

KW - Carica

KW - Circular Dichroism

KW - Latex

KW - Magnetic Resonance Spectroscopy

KW - Peptide Hydrolases

KW - Spectroscopy, Fourier Transform Infrared

KW - Thermogravimetry

KW - X-Ray Diffraction

KW - beta-Cyclodextrins

M3 - Article

VL - 20

SP - 283

EP - 291

JO - BioDrugs

JF - BioDrugs

SN - 1173-8804

IS - 5

ER -