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Effect of process parameters and surfactant additives on the obtained activity of recombinant tryptophan hydroxylase (TPH1) for enzymatic synthesis of 5-hydroxytryptophan (5-HTP).
Vargas, Marta A; Deive, Francisco J; Álvarez, María S; Longo, María A; Rodríguez, Ana; Bernal, Claudia; Martínez, Ronny.
Afiliação
  • Vargas MA; Laboratorio de Tecnología Enzimática para Bioprocesos, Departamento de Ingeniería en Alimentos, Universidad de La Serena, Chile.
  • Deive FJ; Departamento de Ingeniería Química, Campus As Lagoas-Marcosende, Universidade de Vigo, Vigo, Spain; CINTECX - Universidad de Vigo, Campus As Lagoas-Marcosende, Vigo, Spain.
  • Álvarez MS; Departamento de Ingeniería Química, Campus As Lagoas-Marcosende, Universidade de Vigo, Vigo, Spain; CINTECX - Universidad de Vigo, Campus As Lagoas-Marcosende, Vigo, Spain.
  • Longo MA; Departamento de Ingeniería Química, Campus As Lagoas-Marcosende, Universidade de Vigo, Vigo, Spain; CINTECX - Universidad de Vigo, Campus As Lagoas-Marcosende, Vigo, Spain.
  • Rodríguez A; Departamento de Ingeniería Química, Campus As Lagoas-Marcosende, Universidade de Vigo, Vigo, Spain.
  • Bernal C; Laboratorio de Tecnología Enzimática para Bioprocesos, Departamento de Ingeniería en Alimentos, Universidad de La Serena, Chile; Instituto de Investigación Multidisciplinario en Ciencia y Tecnología, Universidad de La Serena, Benavente 980, 1720010 La Serena, Chile.
  • Martínez R; Laboratorio de Tecnología Enzimática para Bioprocesos, Departamento de Ingeniería en Alimentos, Universidad de La Serena, Chile. Electronic address: remartinez@userena.cl.
Enzyme Microb Technol ; 154: 109975, 2022 Mar.
Article em En | MEDLINE | ID: mdl-34952363
5-hydroxytryptophan (5-HTP) is an intermediate molecule in the biosynthesis of serotonin, an important neurotransmitter, regulating a series of metabolic and psychological functions in humans. In this work, we studied the heterologous production of Human tryptophan hydroxylase (TPH1) in Escherichia coli, for the synthesis of 5-hydroxytryptophan (5-HTP) from Tryptophan (Trp). To quantify TPH1 activity, a simple fluorescence-based microtiter plate assay was established, based on the changes in fluorescence emission at 340 nm between substrate and product when excited at 310 nm, allowing quick and reliable quantification of released 5-HTP. To increase enzyme production, heterologous TPH1 production was studied in stirred tank bioreactor scale. The effect of rate of aeration (0.25, 0.50 and 0.75 vvm) and agitation (150, 250 and 500 rpm) was evaluated for biomass production, pH, volumetric oxygen transfer coefficient (kLa) and volumetric TPH1 activity. We determined that high agitation and low aeration allowed reaching the maximum measured enzyme activity. Under such conditions, we observed a 90% substrate conversion, obtaining 90 µM (~0.02 g/L) 5-HTP from a 100 µM Tryptophan substrate solution. Finally, we observed that the addition of Tween 20 (0.1%) in the culture broth under production conditions expanded the pH operation range of TPH1. Our results establish a base for a biocatalytic approach as a potential alternative process for the synthesis of 5-HTP using recombinant TPH1.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Triptofano Hidroxilase / 5-Hidroxitriptofano Limite: Humans Idioma: En Revista: Enzyme Microb Technol Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Chile País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Triptofano Hidroxilase / 5-Hidroxitriptofano Limite: Humans Idioma: En Revista: Enzyme Microb Technol Ano de publicação: 2022 Tipo de documento: Article País de afiliação: Chile País de publicação: Estados Unidos