Comparison of Polyester-Degrading Cutinases from Genus<i>Thermobifida</i>

Kawai, Fusako; Thumarat, Uschara; Kitadokoro, Kengo; Waku, Tsuyoshi; Tada, Tomoko; Tanaka, Naoki; Kawabata, Takeshi

doi:10.1021/bk-2013-1144.ch009

Cited by 16 publications

(18 citation statements)

References 15 publications

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“…A calcium-dependent thermal stabilization of the polyester esterase Est119 from Thermobifida alba has been reported previously [12]. The analysis of the crystal structure of Est119 revealed a calcium binding site close to its active site [13]. Est119 shares a sequence identity of 82% and a highly structural similarity with TfCut2 [11].…”

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confidence: 72%

A disulfide bridge in the calcium binding site of a polyester hydrolase increases its thermal stability and activity against polyethylene terephthalate

et al. 2016

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Elevated reaction temperatures are crucial for the efficient enzymatic degradation of polyethylene terephthalate (PET). A disulfide bridge was introduced to the polyester hydrolase TfCut2 to substitute its calcium binding site. The melting point of the resulting variant increased to 94.7 °C (wild‐type TfCut2: 69.8 °C) and its half‐inactivation temperature to 84.6 °C (TfCut2: 67.3 °C). The variant D204C‐E253C‐D174R obtained by introducing further mutations at vicinal residues showed a temperature optimum between 75 and 80 °C compared to 65 and 70 °C of the wild‐type enzyme. The variant caused a weight loss of PET films of 25.0 ± 0.8% (TfCut2: 0.3 ± 0.1%) at 70 °C after a reaction time of 48 h. The results demonstrate that a highly efficient and calcium‐independent thermostable polyester hydrolase can be obtained by replacing its calcium binding site with a disulfide bridge.

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confidence: 72%

A disulfide bridge in the calcium binding site of a polyester hydrolase increases its thermal stability and activity against polyethylene terephthalate

et al. 2016

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“…Although only Cut190 displays both active (Ca 2+ -bound) (PDB IDs: 4WFJ and 4WFK) and inactive (Ca 2+ -free) (PDB ID: 4WFI) states (Miyakawa et al 2015), the active state corresponds to those of Est119 and other cutinases (PDB IDs: 4EB0, 3VIS, 3WYN, 4CG1, 4CG2, 4CG3, and 5LUI, 5LUJ, 5LUK, and 5LUL) (Sulaiman et al 2014; Kitadokoro et al 2012; Kawai et al 2013; Roth et al 2014; Ribitsch et al 2017). The activities and thermal stabilities of these cutinases increase in the presence of Ca 2+ and Mg 2+ (Kawai et al 2013, 2014; Sulaiman et al 2014; Thumarat et al 2012; Then et al 2015; Miyakawa et al 2015). Est119 retains the same overall structure with or without Ca 2+ (PDB IDs: 3WYN and 3VIS).…”

Section: The Role Of Divalent Cations In Catalysismentioning

confidence: 99%

Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields

Kawai

Kawabata

Oda

2019

Appl Microbiol Biotechnol

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Enzymatic hydrolysis of polyethylene terephthalate (PET) has been the subject of extensive previous research that can be grouped into two categories, viz. enzymatic surface modification of polyester fibers and management of PET waste by enzymatic hydrolysis. Different enzymes with rather specific properties are required for these two processes. Enzymatic surface modification is possible with several hydrolases, such as lipases, carboxylesterases, cutinases, and proteases. These enzymes should be designated as PET surface–modifying enzymes and should not degrade the building blocks of PET but should hydrolyze the surface polymer chain so that the intensity of PET is not weakened. Conversely, management of PET waste requires substantial degradation of the building blocks of PET; therefore, only a limited number of cutinases have been recognized as PET hydrolases since the first PET hydrolase was discovered by Müller et al. ( Macromol Rapid Commun 26:1400–1405, 2005 ). Here, we introduce current knowledge on enzymatic degradation of PET with a focus on the key class of enzymes, PET hydrolases, pertaining to the definition of enzymatic requirements for PET hydrolysis, structural analyses of PET hydrolases, and the reaction mechanisms. This review gives a deep insight into the structural basis and dynamics of PET hydrolases based on the recent progress in X-ray crystallography. Based on the knowledge accumulated to date, we discuss the potential for PET hydrolysis applications, such as in designing waste stream management. Electronic supplementary material The online version of this article (10.1007/s00253-019-09717-y) contains supplementary material, which is available to authorized users.

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“…For both Est119 and Est1, optimum temperature and pH were recorded as 50°C and pH 6.0, respectively ( Thumarat et al, 2015 ). Despite the enzymes displaying 98% similarity with each other, Est1 displayed higher activity and thermostability ( Kawai et al, 2013 ). More recently, an Est1 (A68V/T253P) variant was shown to have the highest thermostability when compared with wild-type and mutant Est1 and Est119 enzymes.…”

Section: Microbial Sources Of Pet Hydrolyzing Enzymesmentioning

confidence: 99%

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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Comparison of Polyester-Degrading Cutinases from GenusThermobifida

Cited by 16 publications

References 15 publications

A disulfide bridge in the calcium binding site of a polyester hydrolase increases its thermal stability and activity against polyethylene terephthalate

A disulfide bridge in the calcium binding site of a polyester hydrolase increases its thermal stability and activity against polyethylene terephthalate

Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields

Microbial Polyethylene Terephthalate Hydrolases: Current and Future Perspectives

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