Your browser doesn't support javascript.
loading
AmyJ33, a truncated amylase with improved catalytic properties.
Hernández-Heredia, Sarahi; Peña-Castro, Julián Mario; Aguilar-Uscanga, María Guadalupe; Olvera, Clarita; Nolasco-Hipólito, Cirilo; Del Moral, Sandra.
Afiliación
  • Hernández-Heredia S; National Institute of Technology of Mexico/Veracruz Institute of Technology/Food Research and Development Unit, Ma. Quevedo 2779, CP 91897, Veracruz, Veracruz, Mexico.
  • Peña-Castro JM; Institute of Biotechnology, University of Papaloapan, Circuito Central 200, CP 68301, Tuxtepec, Oaxaca, Mexico.
  • Aguilar-Uscanga MG; National Institute of Technology of Mexico/Veracruz Institute of Technology/Food Research and Development Unit, Ma. Quevedo 2779, CP 91897, Veracruz, Veracruz, Mexico.
  • Olvera C; Institute of Biotechnology, National Autonomous University of Mexico, Av. Universidad 2001, CP 62210, Cuernavaca, Morelos, Mexico.
  • Nolasco-Hipólito C; Institute of Biotechnology, University of Papaloapan, Circuito Central 200, CP 68301, Tuxtepec, Oaxaca, Mexico.
  • Del Moral S; Researcher for Mexico-National Council for Science and Technology, Food Research and Development Unit, National Institute of Technology of Mexico/Veracruz Institute of Technology, Quevedo 2779, CP 91897, Veracruz, Veracruz, Mexico. sandit.dmv@gmail.com.
Biotechnol Lett ; 44(12): 1447-1463, 2022 Dec.
Article en En | MEDLINE | ID: mdl-36326957
Biochemical and kinetic properties are of special interest for the specific applications of α-amylases in industrial sectors such as textile industries, detergents, biofuels and food among others. Therefore, protein engineering is currently directed towards a continuous demand to improve the properties of amylases and thus meet the specific characteristics for various industrial sectors. In the present work, modular protein engineering was performed to improve the biochemical and kinetic properties of AmyJ33r an α-amylase isolated from Bacillus siamensis JJC33M consisting of five domains, A, B, C, D and E (SBD) (Montor-Antonio et al. in 3 Biotech 7:336, 2017. https://doi.org/10.1007/s13205-017-0954-8 ). AmyJ33r is not active on native starch, only showing activity on gelatinized starch. At the C-terminal, AmyJ33r has a starch binding domain (SBD, domain E) belonging to the CBM26 family. In this study, four truncated versions were constructed and expressed in E. coli (AmyJ33-AB, AmyJ33-ABC, AmyJ33-ABCD, and SBD) to determine the role of the A, B, C, D, and E domains in the biochemical behavior of AmyJ33r on starch. Biochemical and kinetic characterization of the truncated versions showed that domain C is essential for catalysis; domain D improved enzyme activity at alkaline pH values, is also involved negatively in thermostability at 40, 50, and 60 °C and its presence favored the production of maltooligosaccharides with a higher degree of polymerization (DP4). E domain have interaction with raw starch, also the deletion of E domain (SBD) favors the affinity for the substrate while the deletion of D domain increased enzyme kcat at the time of product release. In conclusion, AmyJ33-ABC has better kinetic parameters than AmyJ33-ABCD and AmyJ33r, but is less stable than these two enzymes.
Asunto(s)
Palabras clave
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Escherichia coli / Amilasas Idioma: En Revista: Biotechnol Lett Año: 2022 Tipo del documento: Article País de afiliación: México Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Escherichia coli / Amilasas Idioma: En Revista: Biotechnol Lett Año: 2022 Tipo del documento: Article País de afiliación: México Pais de publicación: Países Bajos