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Thermoanaerobacter tengcongensis esterase resists denaturation by urea and sodium dodecyl sulfate
La esterasa de Thermoanaerobacter tengcongensis resiste la desnaturalización por la urea y el dodecilsulfato de sodio
A esterase de Thermoanaerobacter tengcongensis resiste à desnaturação pela uréia e dodecil sulfato de sódio
DOI:
https://doi.org/10.15446/rcciquifa.v52n3.112485Palabras clave:
Thermoanaerobacter tengcongensis, esterase, thermophiles, protein denaturants, para-nitrophenyl dodecanoate (en)Thermoanaerobacter tengcongensis , esterasa , termófilos, desnaturalizantes de proteínas, para-nitrofenil dodecanoato (es)
Thermoanaerobacter tengcongensis, esterase, termófilos, desnaturantes de proteínas, para-nitrofenil dodecanoato (pt)
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Introduction: The broad applications of lipolytic enzymes in various industrial processes have led to increased interest in esterases with distinctive features. Thermophiles are promising source of esterases with inherent thermal and chemical stability. Thermoanaerobacter tengcongensis esterase (TTE) is one of such esterases with thermostable potential, however, its resistance to protein denaturants, detergents and molecular docking studies are yet to be fully characterised. Aim: Therefore, this study investigated the in vitro and in silico effects of urea and sodium dodecyl sulfate on TTE activity. Experimental: TTE activity was determined spectrophotometrically at 405 nm. TTE was active over a pH range of 3.0 to 12.0 and its activity was optimal at alkaline range of 9.0 and 12.0. Results: TTE was found to be most active at 60 °C with the highest thermal stability at the same temperature. Urea at 0.1 to 4.0 mM had a concentration dependent activating effect on TTE; SDS (0.5 to 4.0 mM) had similar effect on the enzyme. Urea at 0.5, 1.0 and 2.0 mM increased maximum reaction rate (Vmax), catalytic constant (Kcat) and Michaelis constant (Km) of TTE. All concentrations of SDS (0.5 to 2.0 mM) investigated increased Vmax and Kcat, while the Km value of TTE reduced in the presence of 1.0 and 2.0 mM SDS. Structural characterization of TTE substantiates the in vitro thermostability claim. The molecular docking analysis revealed that donepezil demonstrated optimal binding with TTE. Conclusion: the findings from this study showed that TTE strongly resists denaturation by optimal concentrations of urea and SDS.
Introducción: Las amplias aplicaciones de las enzimas lipolíticas en diversos procesos industriales han llevado a un mayor interés en esterasas con características distintivas. Los termófilos son una fuente prometedora de esterasas con estabilidad térmica y química inherente. La esterasa de Thermoanaerobacter tengcongensis (TTE) es una de esas esterasas con potencial termoestable; sin embargo, su resistencia a los desnaturalizantes de proteínas, los detergentes y los estudios de acoplamiento molecular aún no se han caracterizado por completo. Objetivo: Por lo tanto, este estudio investigó los efectos in vitro e in silico de la urea y el dodecilsulfato de sodio sobre la actividad del TTE. Parte experimental: la actividad TTE se determinó espectrofotométricamente a 405 nm. El TTE estuvo activo en un rango de pH de 3,0 a 12,0 y su actividad fue óptima en un rango alcalino de 9,0 y 12,0. Resultados: Se encontró que el TTE era más activo a 60 °C y tenía la mayor estabilidad térmica a la misma temperatura. La urea de 0,1 a 4,0 mM tuvo un efecto activador dependiente de la concentración sobre el ETT; SDS (0,5 a 4,0 mM) tuvo un efecto similar sobre la enzima. La urea en concentraciones de 0,5, 1,0 y 2,0 mM aumentó la velocidad de reacción máxima (Vmax), la constante catalítica (Kcat) y la constante de Michaelis (Km) de TTE. Todas las concentraciones de SDS (0,5 a 2,0 mM) investigadas aumentaron Vmax y Kcat, mientras que el valor de Km de TTE se redujo en presencia de SDS 1,0 y 2,0 mM. La caracterización estructural de TTE fundamenta la afirmación de termoestabilidad in vitro. El análisis de acoplamiento molecular reveló que donepezilo demostró una unión óptima con TTE. Conclusión: los hallazgos de este estudio mostraron que el TTE resiste fuertemente la desnaturalización por concentraciones óptimas de urea y SDS.
Introdução: As amplas aplicações de enzimas lipolíticas em diversos processos industriais têm levado ao aumento do interesse em esterases com características distintivas. Os termófilos são fontes promissoras de esterases com estabilidade térmica e química inerente. A esterase de Thermoanaerobacter tengcongensis (TTE) é uma dessas esterases com potencial termoestável, no entanto, sua resistência a desnaturantes de proteínas, detergentes e estudos de acoplamento molecular ainda não foram totalmente caracterizadas. Objetivo: Portanto, este estudo investigou os efeitos in vitro e in silico da uréia e dodecilsulfato de sódio na atividade do TTE. Parte experimental: A atividade do TTE foi determinada espectrofotometricamente a 405 nm. O TTE foi ativo em uma faixa de pH de 3,0 a 12,0 e sua atividade foi ótima na faixa alcalina de 9,0 e 12,0. Resultados: Descobriu-se que o TTE é mais ativo a 60 °C, com maior estabilidade térmica à mesma temperatura. A uréia em concentrações de 0,1 a 4,0 mM teve um efeito ativador dependente da concentração no ETT; SDS (0,5 a 4,0 mM) teve efeito semelhante na enzima. A uréia a 0,5, 1,0 e 2,0 mM aumentou a taxa máxima de reação (Vmax), a constante catalítica (Kcat) e a constante de Michaelis (Km) do TTE. Todas as concentrações de SDS (0,5 a 2,0 mM) investigadas aumentaram Vmax e Kcat, enquanto o valor de Km de TTE reduziu na presença de 1,0 e 2,0 mM de SDS. A caracterização estrutural do TTE fundamenta a alegação de termoestabilidade in vitro. A análise de acoplamento molecular revelou que o donepezil demonstrou ligação ideal ao TTE. Conclusão: os resultados deste estudo mostraram que o TTE resiste fortemente à desnaturação por concentrações ótimas de uréia e SDS.
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