Your browser doesn't support javascript.
loading
Improving the position specificity of Themomyces lanuginosus lipase based on semi-rational design / 生物工程学报
Chinese Journal of Biotechnology ; (12): 3481-3493, 2023.
Artigo em Chinês | WPRIM | ID: wpr-1007971
ABSTRACT
Diacylglycerol (DAG) is an intermediate product in lipid metabolism and plays an important physiological role in human body. It is mainly prepared by hydrolyzing lipid with lipase. However, research on the detection method of 1, 2-diacylglycerol (1, 2-DAG) and 1, 3-diacylglycerol (1, 3-DAG) and catalytic specificity of lipase was not enough, which limits its wide application. To address these challenges, an efficient quantitative detection method was first established for 1, 2-DAG (0.025-0.200 g/L) and 1, 3-DAG (0.025-0.150 g/L) by combining supercritical fluid chromatography with evaporative light scattering detector and optimizing the detection and analysis parameters. Based on the molecular docking between Thermomyces lanuginosus lipase (TLL) and triolein, five potential substrate binding sites were selected for site-specific saturation mutation to construct a mutation library for enzyme activity and position specificity screening. The specificity of sn-1, 3 of the I202V mutant was the highest in the library, which was 11.7% higher than the specificity of the wild type TLL. In summary, the position specificity of TLL was modified based on a semi-rational design, and an efficient separation and detection method of DAG isomers was also established, which provided a reference for the study of the catalytic specificity of lipase.
Assuntos

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Sítios de Ligação / Catálise / Diglicerídeos / Simulação de Acoplamento Molecular / Lipase Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Biotechnology Ano de publicação: 2023 Tipo de documento: Artigo

Similares

MEDLINE

...
LILACS

LIS

Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Sítios de Ligação / Catálise / Diglicerídeos / Simulação de Acoplamento Molecular / Lipase Limite: Humanos Idioma: Chinês Revista: Chinese Journal of Biotechnology Ano de publicação: 2023 Tipo de documento: Artigo