[Expression and characterization of a bifunctional thermal ß-glucosidase IuBgl3 from thermophilic archaeon Infirmifilum uzonense].
Sheng Wu Gong Cheng Xue Bao
; 38(12): 4644-4657, 2022 Dec 25.
Article
em Zh
| MEDLINE
| ID: mdl-36593199
ABSTRACT
ß-glucosidase has important applications in food, medicine, biomass conversion and other fields. Therefore, exploring ß-glucosidase with strong stability and excellent properties is a research hotspot. In this study, a GH3 family ß-glucosidase gene named Iubgl3 was successfully cloned from Infirmifilum uzonense. Sequence analysis showed that the full length of Iubgl3 was 2 106 bp, encoding 702 amino acids, with a theoretical molecular weight of 77.0 kDa. The gene was cloned and expressed in E. coli and the enzymatic properties of purified IuBgl3 were studied. The results showed that the optimal pH and temperature for pNPG hydrolysis were 5.0 and 85 â, respectively. The enzyme has good thermal stability, and more than 85% of enzyme activity can be retained after being treated at 80 â for2 h. This enzyme has good pH stability and more than 85% of its activity can be retained after being treated at pH 4.0-11.0 for 1 h. It was found that the enzyme had high hydrolysis ability to p-nitrophenyl ß-d-glucoside (pNPG) and p-nitrophenyl ß-d-xylopyranoside (pNPX). When pNPG was used as the substrate, the kinetic parameters Km and Vmax were 0.38 mmol and 248.55 µmol/(mg·min), respectively, and the catalytic efficiency kcat/Km was 6 149.20 s-1mmol-1. Most metal ions had no significant effect on the enzyme activity of IuBgl3. SDS completely inactivated the enzyme, while EDTA increased the enzyme activity by 30%. This study expanded the ß-glucosidase gene diversity of the thermophilic archaea GH3 family and obtained a thermostable acid bifunctional enzyme with good industrial application potential.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Contexto em Saúde:
3_ND
Problema de saúde:
3_neglected_diseases
/
3_zoonosis
Assunto principal:
Archaea
/
Beta-Glucosidase
Idioma:
Zh
Revista:
Sheng Wu Gong Cheng Xue Bao
Assunto da revista:
BIOTECNOLOGIA
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
China