Purification and Characterization of Four New Cysteine Endopeptidases From Fruits of Bromelia pinguin L. Grown in Cuba

Payrol, Juan Abreu; Obregón, Walter David; Trejo, Sebastián A.; Caffini, Néstor Oscar

doi:10.1007/s10930-007-9111-2

Cited by 25 publications

(16 citation statements)

References 21 publications

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“…4B); this behavior may be due to the fact that DTT is a reducing agent of sulfhydryl groups, which is used to eliminate the disulfide bonds between one or more polypeptide chains. However, it is also known that at low concentrations, DTT protects the proteolytic activity of extracts of B. pinguin [23], because it favors the formation of sulfhydryl group (-SH), allowing the active site of the enzyme to be active. It is important to note that similar effects of DTT on B. karatas were observed in this study; one could consider that its proteases are also cysteine proteases.…”

Section: Discussionmentioning

confidence: 99%

Isoelectric focusing, effect of reducing agents and inhibitors: partial characterization of proteases extracted from Bromelia karatas

et al. 2018

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The aim of this research is the partial characterization of proteases extracted from B. karatas; the isolation and purification of proteases from B. karatas fruits were achieved using precipitation, separation by size exclusion chromatography and anion-exchange chromatography; molecular mass (MM) was determined, and the effect of inhibitors, reducing agents and heat on enzyme activity was analyzed. These proteases were compared with proteases from Bromelia pinguin (B. pinguin) and evaluated under similar conditions. The isolation procedure was adequate; only a few protein bands are present in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Furthermore, zymogram analysis showed protein bands with enzyme activity. Inhibitors, reducing agents and heat were unable to inactivate the proteases extracted from B. karatas and B. pinguin. The semi-purified extracts are a set of proteases with a MM of 66 kDa, but different isoelectric points (3.5-6.5 for B. karatas and 5-9 for B. pinguin), which are found in quaternary structures with proteolytic activity. When denatured, they segment into fragments of approximately 20 and 10 kDa. The data indicate that these plants could be used as sources of proteases since they present good proteolytic activity (21.93 U T for proteases from B. karatas and 43.58 U T for proteases from B. pinguin) and that B. Karatas has potential applications comparable to B. pinguin in the food and health industries.

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Section: Discussionmentioning

confidence: 99%

Isoelectric focusing, effect of reducing agents and inhibitors: partial characterization of proteases extracted from Bromelia karatas

et al. 2018

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“…It has been reported that the different genetic characteristics of each fruit species synthetize endopeptidases with different structure and catalytic activities (Headon and Walsh, 1994). B. karatas proteases had the lowest enzymatic activity; but the proteolytic activity was similar than purified B. pinguin proteases (7.11 to 11.0 U/mg protein) of fruits harvested in Cuba (Payrol et al, 2005(Payrol et al, , 2008.…”

Section: Proteolytic Activitymentioning

confidence: 99%

Antioxidant capacity of egg, milk and soy protein hydrolysates and biopeptides produced by Bromelia pinguin and Bromelia karatas-derived proteases

et al. 2018

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Given the lack of knowledge in the generation and biological activity of biopeptides (BP) produced by Bromelia pinguin and Bromelia karatas-derived proteases, the objective of this work was to evaluate the antioxidant capacity (AOX) of protein hydrolysates and peptide fractions from hydrolysis of egg, milk and soy proteins with enzymes extracted from Bromelia pinguin and Bromelia karatas fruits. The degree of hydrolysis (DH) of the mentioned proteins was measured. The hydrolysates were sequentially fractionated by ultrafiltration using different membrane pore sizes (≤30, ≤10, ≤5 and ≤1 kDa) to obtain biopeptide fractions (BPF). The antioxidant capacity (AOX) of protein hydrolysates and BPF were evaluated. AOX was performed using 2,2´-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS•+), ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) assays. after the enzymatic action of B. pinguin and B. karatas proteases, the highest DH values for egg, milk and soy protein hydrolysates ranged from 50-61%, 55-68% and 73-81%, respectively. The AOX was higher for soy hydrolysates than for egg and milk hydrolysates. Additionally, the highest AOX from ABTS and FRAP assays was observed for BPF ≤30 kDa, PF ≤5 kDa and ≤1 kDa, respectively. These results highlight the potential of B. pinguin and B. karatas proteases to hydrolyze food proteins and release bioactive peptides. Moreover, it has been shown that the hydrolysis products are a mixture of BP of different molecular weights, which could present AOX through different mechanisms.

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“…Sin embargo, hay que considerar aquí la presencia en B. pinguín de enzimas proteasas, en particular la pinguinaína, un pre parado enzimático con actividad proteolítica presente en el fruto de esta especie la cual ya ha sido blanco de diferentes estudios (27), (28), (29), (9), (30).…”

Section: Actividad Antifúngicaunclassified

CARACTERIZACIÓN FITOQUÍMICA DEL EXTRACTO HIDROALCOHÓLICO DEL FRUTO DE Bromelia pinguin Y EVALUACIÓN DE SU ACTIVIDAD ANTIFÚNGICA FRENTE A UN HONGO AISLADO DEL FRUTO DE CACAO (Theobroma cacao L.) EN ESTADO DE PUDRICIÓN.

Vargas

Jaramillo

Guzmán

et al. 2018

Rev. Invest. Univ. Quindío

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Se evaluó el efecto antifúngico del extracto hidroalcohólico de fruto de B. pinguin frente a un hongo aislado de frutos enfermos en estado de pudrición de Cacao (Theobroma cacao L.) mediante un ensayo de inhibición de crecimiento micelial empleando el método de dilución en agar. El extracto tuvo un efecto antifúngico bastante notorio en un rango de concentraciones entre 2,5 y 10 % (V/V) y se pudo evidenciar además una fuerte relación dosis-respuesta. Un análisis fitoquímico preliminar, permitió identificar flavonoides, antraquinonas, taninos, saponinas y glucósidos cardiotónicos como los principales metabolitos secundarios presentes en el extracto hidroalcohólico de B. pinguin; sin embargo, aquí se discute sobre la Pinguinaína, como principal responsable de dicha actividad.

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Purification and Characterization of Four New Cysteine Endopeptidases From Fruits of Bromelia pinguin L. Grown in Cuba

Cited by 25 publications

References 21 publications

Isoelectric focusing, effect of reducing agents and inhibitors: partial characterization of proteases extracted from Bromelia karatas

Isoelectric focusing, effect of reducing agents and inhibitors: partial characterization of proteases extracted from Bromelia karatas

Antioxidant capacity of egg, milk and soy protein hydrolysates and biopeptides produced by Bromelia pinguin and Bromelia karatas-derived proteases

CARACTERIZACIÓN FITOQUÍMICA DEL EXTRACTO HIDROALCOHÓLICO DEL FRUTO DE Bromelia pinguin Y EVALUACIÓN DE SU ACTIVIDAD ANTIFÚNGICA FRENTE A UN HONGO AISLADO DEL FRUTO DE CACAO (Theobroma cacao L.) EN ESTADO DE PUDRICIÓN.

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