Engineering the plastic degradation enzyme Ple629 from marine consortium to improve its thermal stability.
- Author:
Yipei ZHAO
1
;
Hao WANG
2
;
Pan WU
1
;
Zhishuai LI
2
;
Fufeng LIU
1
;
Qun GU
2
;
Weidong LIU
2
;
Jian GAO
2
;
Xu HAN
2
Author Information
- Publication Type:Journal Article
- Keywords: expression and purification; hydrolase; mutagenesis; polybutylene adipate terephthalate; polyethylene terephthalate; thermal stability
- MeSH: Plastics/metabolism*; Polyethylene Terephthalates/metabolism*; Biodegradation, Environmental; Metagenome
- From: Chinese Journal of Biotechnology 2023;39(5):2040-2052
- CountryChina
- Language:Chinese
- Abstract: Petrochemical-derived polyester plastics such as polyethylene terephthalate (PET) and polybutylene adipate terephthalate (PBAT) have been widely used. However, the difficulty to be degraded in nature (PET) or the long biodegradation cycle (PBAT) resulted in serious environmental pollution. In this connection, treating these plastic wastes properly becomes one of the challenges of environment protection. From the perspective of circular economy, biologically depolymerizing the waste of polyester plastics and reusing the depolymerized products is one of the most promising directions. Recent years have seen many reports on polyester plastics degrading organisms and enzymes. Highly efficient degrading enzymes, especially those with better thermal stability, will be conducive to their application. The mesophilic plastic-degrading enzyme Ple629 from the marine microbial metagenome is capable of degrading PET and PBAT at room temperature, but it cannot tolerate high temperature, which hampers its potential application. On the basis of the three-dimensional structure of Ple629 obtained from our previous study, we identified some sites which might be important for its thermal stability by structural comparison and mutation energy analysis. We carried out transformation design, and performed expression, purification and thermal stability determination of the mutants. The melting temperature (Tm) values of mutants V80C and D226C/S281C were increased by 5.2 ℃ and 6.9 ℃, respectively, and the activity of mutant D226C/S281C was also increased by 1.5 times compared with that of the wild-type enzyme. These results provide useful information for future engineering and application of Ple629 in polyester plastic degradation.
