Rational design of L-arabinose isomerase from Lactobacillus fermentum and its application in D-tagatose production / 生物工程学报
Chinese Journal of Biotechnology
; (12): 1107-1118, 2023.
Artigo
em Chinês
| WPRIM (Pacífico Ocidental)
| ID: wpr-970426
Biblioteca responsável:
WPRO
ABSTRACT
L-arabinose isomerase (L-AI) is the key enzyme that isomerizes D-galactose to D-tagatose. In this study, to improve the activity of L-arabinose isomerase on D-galactose and its conversion rate in biotransformation, an L-arabinose isomerase from Lactobacillus fermentum CGMCC2921 was recombinantly expressed and applied in biotransformation. Moreover, its substrate binding pocket was rationally designed to improve the affinity and catalytic activity on D-galactose. We show that the conversion of D-galactose by variant F279I was increased 1.4 times that of the wild-type enzyme. The Km and kcat values of the double mutant M185A/F279I obtained by superimposed mutation were 530.8 mmol/L and 19.9 s-1, respectively, and the catalytic efficiency was increased 8.2 times that of the wild type. When 400 g/L lactose was used as the substrate, the conversion rate of M185A/F279I reached a high level of 22.8%, which shows great application potential for the enzymatic production of tagatose from lactose.
Texto completo:
Disponível
Base de dados:
WPRIM (Pacífico Ocidental)
Assunto principal:
Aldose-Cetose Isomerases
/
Limosilactobacillus fermentum
/
Galactose
/
Hexoses
/
Concentração de Íons de Hidrogênio
/
Lactose
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2023
Tipo de documento:
Artigo