Semi-rational design improves the catalytic activity of phenylalanine ammonia lyase from Anabaena variabilis
10.16438/j.0513-4870.2022-0631
- VernacularTitle:半理性设计提高Anabaena variabilis来源的苯丙氨酸解氨酶的催化活性
- Author:
Xi-yu WEI
1
;
Cui-yue FENG
1
;
Rui-jie LV
1
;
Shuai FAN
2
;
Zhao-yong YANG
2
;
Zhi-fei ZHANG
1
Author Information
1. School of Pharmacy, North China University of Science and Technology, Tangshan 063200, China
2. Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China
- Publication Type:Research Article
- Keywords:
phenylalanine ammonia lyase;
saturation mutation;
catalytic efficiency;
molecular docking
- From:
Acta Pharmaceutica Sinica
2022;57(12):3669-3674
- CountryChina
- Language:Chinese
-
Abstract:
Phenylalanine ammonia lyase (PAL) can catalyze L-phenylalanine to produce trans-cinnamic acid, which is widely used in the fields of pharmacy, food and agriculture. In particular, phenylalanine ammonia lyase from Anabaena variabilis (AvPAL) is the only protein drug for the treatment of phenylketonuria. However, the poor activity and low stability limit the application in industry of AvPAL. In this study, the key amino acids of substrate-binding cavity in AvPAL were identified by screening the single site saturation mutagenesis library. Subsequently, the impact of replacing M222 with the additional 19 amino acids on activity was also evaluated by site-directed mutagenesis. It was found that the kcat values of mutants M222L and M222V were 90% and 60% higher than that of AvPAL, and the kcat/Km was 1.4 and 1.5 times as that of AvPAL. Molecular docking results revealed that the higher activity of M222L and M222V may be due to the increase of hydrophobicity favorable for the substrate-binding cavity. This study is important for elucidating the structure-function relationship of AvPAL.
