Structural bioinformatics-based protein engineering of thermo-stable PETase from Ideonella sakaiensis

Son, Hyeoncheol Francis; Joo, Seongjoon; Seo, Hogyun; Sagong, Hye-Young; Lee, Seul Hoo; Hong, Hwaseok; Kim, Kyungjin

doi:10.1016/j.enzmictec.2020.109656

Cited by 92 publications

(59 citation statements)

References 39 publications

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“…Upon comparison of the mutant with wild-type IsPETase, degradative activity toward PET film was shown to be 14-fold higher at 40 • C. In addition, the melting temperature of this variant increased by 8.81 • C and high thermostability was further verified by a heat-inactivation experiment (Son et al, 2019). The same group have since introduced mutations N246D and S242T, generating a quadruple IsPETase variant which displayed a 58fold increase in hydrolyzing activity over its wild-type at 37 • C, and maintained activity under these conditions for 20 days (Son et al, 2020).…”

Section: Enzyme Properties Catalytic Efficiency and Thermal Stabilitymentioning

confidence: 90%

Microbial Polyethylene Terephthalate Hydrolases: Current and Future Perspectives

2020

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Plastic has rapidly transformed our world, with many aspects of human life now relying on a variety of plastic materials. Biological plastic degradation, which employs microorganisms and their degradative enzymes, has emerged as one way to address the unforeseen consequences of the waste streams that have resulted from mass plastic production. The focus of this review is microbial hydrolase enzymes which have been found to act on polyethylene terephthalate (PET) plastic. The best characterized examples are discussed together with the use of genomic and protein engineering technologies to obtain PET hydrolase enzymes for different applications. In addition, the obstacles which are currently limiting the development of efficient PET bioprocessing are presented. By continuing to study the possible mechanisms and the structural elements of key enzymes involved in microbial PET hydrolysis, and by assessing the ability of PET hydrolase enzymes to work under practical conditions, this research will help inform large-scale waste management operations. Finally, the contribution of microbial PET hydrolases in creating a potential circular PET economy will be explored. This review combines the current knowledge on enzymatic PET processing with proposed strategies for optimization and use, to help clarify the next steps in addressing pollution by PET and other plastics.

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Section: Enzyme Properties Catalytic Efficiency and Thermal Stabilitymentioning

confidence: 90%

Microbial Polyethylene Terephthalate Hydrolases: Current and Future Perspectives

2020

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“…Structural biology data combined with computer methods for protein design have been mainly focused in the improvement of protein stability. Son and coworkers applied a rational protein engineering method based on sequence homology to design a I. sakaiensis PETase mutant (S121E-D186H-S242T-N246D) that showed prolonged enzymatic activity over 20 days [ 59 ]. Moreover, Cui and coworkers developed a computational strategy for the mutational analysis of PETase and the design of a multiple mutant with enhanced thermostability when compared with the wild-type protein.…”

Section: Discussionmentioning

confidence: 99%

Structural Insights into Carboxylic Polyester-Degrading Enzymes and Their Functional Depolymerizing Neighbors

Leitão

Enguita

2021

IJMS

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Esters are organic compounds widely represented in cellular structures and metabolism, originated by the condensation of organic acids and alcohols. Esterification reactions are also used by chemical industries for the production of synthetic plastic polymers. Polyester plastics are an increasing source of environmental pollution due to their intrinsic stability and limited recycling efforts. Bioremediation of polyesters based on the use of specific microbial enzymes is an interesting alternative to the current methods for the valorization of used plastics. Microbial esterases are promising catalysts for the biodegradation of polyesters that can be engineered to improve their biochemical properties. In this work, we analyzed the structure-activity relationships in microbial esterases, with special focus on the recently described plastic-degrading enzymes isolated from marine microorganisms and their structural homologs. Our analysis, based on structure-alignment, molecular docking, coevolution of amino acids and surface electrostatics determined the specific characteristics of some polyester hydrolases that could be related with their efficiency in the degradation of aromatic polyesters, such as phthalates.

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“…Introduction of disulfide bonds or salt bridges can be beneficial to the enhancement of the thermostability of plastic-degrading enzymes (Figure 2A) [44][45][46][47]. Disulfide bonds and salt bridges are crucial for protein folding with the correct local or global conformation that could confer thermal resistance.…”

Section: Protein Engineering Of Plastic-degrading Enzymesmentioning

confidence: 99%

“…For example, the D204C and E253C mutations at the calcium binding site of an esterase TfCut2 from Thermobifida fusca Trends in Biotechnology formed a disulfide bond, considerably increasing the protein melting temperature and plastic hydrolysis activity [48]. Additionally, the formation of a salt bridge between the negativelycharged N246D residue and positively-charged Arg280 residue might contribute to the improved thermostability of the engineered PETase N246D [45]. Moreover, disulfide bonds and salt bridge construction could work synergistically to further benefit the thermostability of the enzyme [48].…”

Section: Protein Engineering Of Plastic-degrading Enzymesmentioning

confidence: 99%

“…To avoid such negative impact, profound understanding and analyses of enzyme structure-function relationships are needed. In fact, in most of the published studies, enzymes engineered with enhanced thermostability also had improved or unchanged plastic degradation efficiency [45,46,49,54,58]. For example, the engineered LCC variants exhibited both enhanced thermostability and increased PET degradation efficiency compared with wild-type LCC [58].…”

Section: Trends Trends In In Biotechnology Biotechnologymentioning

confidence: 99%

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Enzyme discovery and engineering for sustainable plastic recycling

Zhu

Wang

Wei

2022

Trends in Biotechnology

204

114

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The drastically increasing amount of plastic waste is causing an environmental crisis that requires innovative technologies for recycling post-consumer plastics to achieve waste valorization while meeting environmental quality goals. Biocatalytic depolymerization mediated by enzymes has emerged as an efficient and sustainable alternative for plastic treatment and recycling. A variety of plasticdegrading enzymes have been discovered from microbial sources. Meanwhile, protein engineering has been exploited to modify and optimize plastic-degrading enzymes. This review highlights the recent trends and up-to-date advances in mining novel plastic-degrading enzymes through state-of-the-art omics-based techniques and improving the enzyme catalytic efficiency and stability via various protein engineering strategies. Future research prospects and challenges are also discussed. Biocatalysis as an Emerging Solution for the Global Plastic Waste ChallengePlastic materials play a revolutionary role in the modern world, although the enormous manufacture and extensive use of plastic commodities inevitably generate an extraordinary amount of post-consumer plastic waste. Around 12 000 million metric tons of plastic waste are predicted to accumulate in landfills and the natural environment by 2050 [1]. Improper handling of plastic waste has caused a grand environmental challenge. The debris of plastic waste, especially microplastics (see Glossary), can impose hazardous effects on various organisms and eventually threaten human well-being [2-5]. In addition, the degradation resistance of plastics further escalates their adverse environmental impacts [6]. Therefore, it is urgent to develop innovative technologies for treatment and recycling of post-consumer plastics, to achieve both waste valorization and environmental protection.Enzymatic biocatalysis has gained increasing attention as an eco-friendly alternative to conventional plastic treatment and recycling methods (Box 1) [7]. To date, various microbial plastic-degrading enzymes have been discovered, representing promising biocatalyst candidates for plastic depolymerization. Considering the ubiquity of plastics in different ecosystems and the tremendous metabolic and genetic diversity of microorganisms, microbial communities in various habitats have likely evolved capabilities in plastic decomposition and utilization. The plastic-degrading enzymes identified so far might only account for a small portion of the enzymes relevant to plastic depolymerization in the environment. Therefore, it is of ever-growing interest to explore diverse environments to discover new plastic-degrading enzymes with desirable properties and functionalities. However, naturally occurring plasticdegrading enzymes are not well suited for synthetic plastic degradation in industrial applications due to poor thermostability and low catalytic activity. Particularly, synthetic plastic materials usually possess distinct physical and chemical properties (e.g., high crystallinity) that render them more resistant to...

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Structural bioinformatics-based protein engineering of thermo-stable PETase from Ideonella sakaiensis

Cited by 92 publications

References 39 publications

Microbial Polyethylene Terephthalate Hydrolases: Current and Future Perspectives

Microbial Polyethylene Terephthalate Hydrolases: Current and Future Perspectives

Structural Insights into Carboxylic Polyester-Degrading Enzymes and Their Functional Depolymerizing Neighbors

Enzyme discovery and engineering for sustainable plastic recycling

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