Research Advance in Polyethylene Terephthalate Hydrolytic Enzymes

doi:10.13560/j.cnki.biotech.bull.1985.2020-0336

Abstract

Abstract:

Polyethylene terephthalate（PET）is one of the most widely used plastics and accumulation of the discarded PET has become a global environmental issue. PET hydrolytic enzymes mainly hydrolyze PET into soluble building blocks：terephthalic acid（TPA）and ethylene glycol（EG）at moderate conditions. Enzymatic degradation of the recalcitrant plastic PET may provide a more environmentally friendly approach to solve the problem of “white pollution.” Here,we reviewed the 3D structures,catalytic mechanisms and PET degradation activity of four PET hydrolases,including cutinases,PETase,lipases and MHETase,mainly focusing on the protein engineering effort of cutinases and PETase. We also discussed the key scientific challenges on the development of high efficient enzymatic degradation of PET,aiming to provide theoretical basis for the engineering of more efficient PET hydrolytic enzymes.

Key words: polyethylene terephthalate（PET）, cutinases, PETase, lipases, MHETase, catalytic mechanism, protein engineering

SHI Li-xia, GAO Song-feng, ZHU Lei-lei. Research Advance in Polyethylene Terephthalate Hydrolytic Enzymes[J]. Biotechnology Bulletin, 2020, 36(10): 226-236.

Figures/Tables 7

References 78

[1]	Webb HK, Arnott J, Crawford RJ, et al. Plastic degradation and its environmental implications with special reference to poly(ethylene terephthalate)[J]. Polymers, 2013,5(1):1-18.
[2]	Bornscheuer UT. Feeding on plastic[J]. Science, 2016,351(6278):1154-1155.
[3]	李秀华, 王自瑛. PET降解研究进展[J]. 塑料科技, 2011,39(4):110-114.
	Li XH, Wang ZY. Research progress on degradation of PET[J]. Plastics Science and Technology, 2011,39(4):110-114.
[4]	Sulaiman S, You DJ, Kanaya E, et al. Crystal structure and thermodynamic and kinetic stability of metagenome-derived LC-cutinase[J]. Biochemistry, 2014,53(11):1858-1869. URL pmid: 24593046
[5]	Shirke AN, White C, Englaender JA, et al. Stabilizing leaf and branch compost cutinase(LCC)with glycosylation:mechanism and effect on PET hydrolysis[J]. Biochemistry, 2018,57(7):1190-1200. URL pmid: 29328676
[6]	Ronkvist ÅM, Xie W, Lu W, et al. Cutinase-catalyzed hydrolysis of poly(ethylene terephthalate)[J]. Macromolecules, 2009,42(14):5128-5138.
[7]	Kold D, Dauter Z, Laustsen AK, et al. Thermodynamic and structural investigation of the specific SDS binding of Humicola insolens cutinase[J]. Protein Sci, 2014,23(8):1023-1035. URL pmid: 24832484
[8]	Sonia L, Czjzek M, Lamzin V, et al. Atomic resolution(1. 0 Å)crystal structure of Fusarium solani cutinase:stereochemical analysis[J]. Journal of Molecular Biology, 1997,268(4):779-799. URL pmid: 9175860
[9]	Acero EH, Ribitsch D, Steinkellner G, et al. Enzymatic surface hydrolysis of PET:effect of structural diversity on kinetic properties of cutinases from Thermobifida[J]. Macromolecules, 2011,44(12):4632-4640.
[10]	Ribitsch D, Hromic A, Zitzenbacher S, et al. Small cause, large effect:structural characterization of cutinases from Thermobifida cellulosilytica[J]. Biotechnol Bioeng, 2017,114(11):2481-2488. URL pmid: 28671263
[11]	Then J, Wei R, Oeser T, et al. Ca²+ and Mg²+ binding site engineering increases the degradation of polyethylene terephthalate films by polyester hydrolases from Thermobifida fusca[J]. Biotechnol J, 2015,10(4):592-598. URL pmid: 25545638
[12]	Roth C, Wei R, Oeser T, et al. Structural and functional studies on a thermostable polyethylene terephthalate degrading hydrolase from Thermobifida fusca[J]. Applied Microbiology & Biotechnology, 2014,98(18):7815-7823.
[13]	Almansa E, Heumann S, Eberl A, et al. Enzymatic surface hydrolysis of PET enhances bonding in PVC coating[J]. Biocatalysis and Biotransformation, 2009,26(5):365-370.
[14]	Brueckner T, Eberl A, Heumann S, et al. Enzymatic and chemical hydrolysis of poly(ethylene terephthalate)fabrics[J]. Journal of Polymer Science Part A:Polymer Chemistry, 2008,46(19):6435-6443.
[15]	Eberl A, Heumann S, Brückner T, et al. Enzymatic surface hydrolysis of poly(ethylene terephthalate)and bis(benzoyloxyethyl)terephthalate by lipase and cutinase in the presence of surface active molecules[J]. Journal of Biotechnology, 2009,143(3):207-212. URL pmid: 19616594
[16]	Eberl A, Heumann S, Kotek R, et al. Enzymatic hydrolysis of PTT polymers and oligomers[J]. Journal of Biotechnology, 2008,135(1):45-51. URL pmid: 18405994
[17]	Fischer-Colbrie G, Heumann S, Liebminger S, et al. New enzymes with potential for PET surface modification[J]. Biocatalysis, 2009,22(5-6):341-346.
[18]	Hsieh YL, Cram LA. Enzymatic hydrolysis to improve wetting and absorbency of polyester fabrics[J]. Textile Research Journal, 2016,68(5):311-319.
[19]	Kim HR, Song WS. Optimization of enzymatic treatment of polyester fabrics by lipase from Porcine Ppancreas[J]. Fibers and Polymers, 2008,9(4):423-430.
[20]	Kim HR, Song WS. Lipase treatment of polyester fabrics[J]. Fibers & Polymers, 2006,7(4):339-343.
[21]	Lee SH, Song WS. Surface modification of polyester fabrics by enzyme treatment[J]. Fibers & Polymers, 2010,11(1):54-59.
[22]	Mccloskey JS. Method of treating polyester fabrics：US, 20060042019A1[P]. 2006-03-02.
[23]	Yoshida S, Hiraga K, Takehana T, et al. A bacterium that degrades and assimilates poly(ethylene terephthalate)[J]. Science, 2016,351(6278):1196-1199.
[24]	Purdy RE, Kolattukudy PE. Hydrolysis of plant cuticle by plant pathogens. Properties of cutinase I, cutinase II, and a nonspecific esterase isolated from Fusarium solani pisi[J]. Biochemistry, 1975,14(13):2832-2840.
[25]	Silva C, Da S, Silva N, et al. Engineered Thermobifida fusca cutinase with increased activity on polyester substrates[J]. Biotechnology Journal, 2011,6(10):1230-1239. doi: 10.1002/biot.201000391 URL pmid: 21751386
[26]	Araújo R, Silva C, O’Neill A, et al. Tailoring cutinase activity towards polyethylene terephthalate and polyamide 6, 6 fibers[J]. Journal of Biotechnology, 2007,128(4):849-857. URL pmid: 17306400
[27]	李秀, 杨海涛, 王泽方. 聚对苯二甲酸乙二醇酯降解酶的研究进展[J]. 微生物学报, 2019,59(12):2251-2262.
	Li X, Yang HT, Wang ZF. Advance in polyethylene terephthalate degrading enzyme[J]. Acta Microbiologica Sinica, 2019,59(12):2251-2262.
[28]	Pellis A, Haernvall K, Pichler CM, et al. Enzymatic hydrolysis of poly(ethylene furanoate)[J]. Journal of Biotechnology, 2016,235:47-53. URL pmid: 26854948
[29]	Wei R, Zimmermann W. Biocatalysis as a green route for recycling the recalcitrant plastic polyethylene terephthalate[J]. Microb Biotechnol, 2017,10(6):1302-1307. URL pmid: 28401691
[30]	Kawabata T, Oda M, Kawai F. Mutational analysis of cutinase-like enzyme, Cut190, based on the 3D docking structure with model compounds of polyethylene terephthalate[J]. Journal of Bioscience and Bioengineering, 2017,124(1):28-35. URL pmid: 28259444
[31]	Then J, Wei R, Oeser T, et al. A disulfide bridge in the calcium binding site of a polyester hydrolase increases its thermal stability and activity against polyethylene terephthalate[J]. FEBS Open Bio, 2016,6(5):425-432. doi: 10.1002/2211-5463.12053 URL
[32]	Kim HW, Ishikawa K. The role of disulfide bond in hyperthermo-philic endocellulase[J]. Extremophiles, 2013,17(4):593-599. doi: 10.1007/s00792-013-0542-8 URL pmid: 23624891
[33]	Han ZL, Han SY, Zheng SP, et al. Enhancing thermostability of a Rhizomucor mieheilipase by engineering a disulfide bond and displaying on the yeast cell surface[J]. Appl Microbiol Biotechnol, 2009,85(1):117-126. doi: 10.1007/s00253-009-2067-8 URL pmid: 19533118
[34]	Tournier V, Topham CM, Gilles A, et al. An engineered PET depolymerase to break down and recycle plastic bottles[J]. Nature, 2020,580(7802):216-219. URL pmid: 32269349
[35]	Barth M, Oeser T, Wei R, et al. Effect of hydrolysis products on the enzymatic degradation of polyethylene terephthalate nanoparticles by a polyester hydrolase from Thermobifida fusca[J]. Biochemical Engineering Journal, 2015,93:222-228.
[36]	Wei R, Oeser T, Schmidt J, et al. Engineered bacterial polyester hydrolases efficiently degrade polyethylene terephthalate due to relieved product inhibition[J]. Biotechnology & Bioengineering, 2016,113(8):1658-1665. URL pmid: 26804057
[37]	Barth M, Honak A, Oeser T, et al. A dual enzyme system composed of a polyester hydrolase and a carboxylesterase enhances the biocatalytic degradation of polyethylene terephthalate films[J]. Biotechnology Journal, 2000,11(8):1082-1087. URL pmid: 27214855
[38]	Hachem MA, Karlsson EN, Bartonek-Roxâ E, et al. Carbohydrate-binding modules from a thermostable Rhodothermus marinus xylanase:cloning, expression and binding studies[J]. Biochem J, 2000,345(Pt 1):53-60.
[39]	Bolam DN, Ciruela A, McQueen-Mason S, et al. Pseudomonas cellulose-binding domains mediate their effects by increasing enzyme substrate proximity[J]. The Biochemical journal, 1998,331(Pt 3):775-781.
[40]	Levy I, Shoseyov O. Cellulose-binding domains:Biotechnological applications[J]. Biotechnology Advances, 2002,20(3):191-213.
[41]	Ribitsch D, Yebra AO, Zitzenbacher S, et al. Fusion of binding domains to Thermobifida cellulosilytica cutinase to tune sorption characteristics and enhancing PET hydrolysis[J]. Biomacromolecules, 2013,14(6):1769-1776. URL pmid: 23718548
[42]	Ribitsch D, Acero EH, Przylucka A, et al. Enhanced cutinase-catalyzed hydrolysis of polyethylene terephthalate by covalent fusion to hydrophobins[J]. Appl Environ Microbiol, 2015,81(11):3586-3592. URL pmid: 25795674
[43]	Joo S, Cho IJ, Seo H, et al. Structural insight into molecular mechanism of poly(ethylene terephthalate)degradation[J]. Nature Communications, 2018,9(1):382.
[44]	Han X, Liu W, Huang JW, et al. Structural insight into catalytic mechanism of PET hydrolase[J]. Nat Commun, 2017,8(1):1-6. URL pmid: 28232747
[45]	Chen CC, Han X, Ko TP, et al. Structural studies reveal the molecular mechanism of PETase[J]. The FEBS journal, 2018,285(20):3717-3723. URL pmid: 30048043
[46]	Fecker T, Galaz-Davison P, Engelberger F, et al. Active site flexibility as a hallmark for efficient PET degradation by I. sakaiensis PETase[J]. Biophys J, 2018,114(6):1302-1312. URL pmid: 29590588
[47]	Buller AR, Townsend CA. Intrinsic evolutionary constraints on protease structure, enzyme acylation, and the identity of the catalytic triad[J]. Proc Natl Acad Sci USA, 2013,110(8):E653-E661. URL pmid: 23382230
[48]	靳玉瑞, 李爱秀, 张力. 一种新型PET水解酶的结构与催化机制研究进展[J]. 中国塑料, 2019,33(3):106-112.
	Jin YR, Li AX, Zhang L. Progress of the structure and catalytic mechanism of one new PET hydrolase[J]. China Plastics, 2019,33(3):106-112.
[49]	Papadopoulou A, Hecht K, Buller R. Enzymatic PET degradation[J]. Chimia, 2019,73(9):743-749. URL pmid: 31514776
[50]	Wei R, Song C, Gräsing D, et al. Conformational fitting of a flexible oligomeric substrate does not explain the enzymatic PET degradation[J]. Nature Communications, 2019,10(1):5581. doi: 10.1038/s41467-019-13492-9 URL
[51]	Seo H, Cho IJ, Joo S, et al. Reply to “Conformational fitting of a flexible oligomeric substrate does not explain the enzymatic PET degradation”[J]. Nature Communications, 2019,10(1):5582. URL pmid: 31811201
[52]	Taniguchi I, Yoshida S, Hiraga K, et al. Biodegradation of PET:current status and application aspects[J]. ACS Catalysis, 2019,9(5):4089-4105. doi: 10.1021/acscatal.8b05171 URL
[53]	Wei R, Oeser T, Barth M, et al. Turbidimetric analysis of the enzymatic hydrolysis of polyethylene terephthalate nanoparticles[J]. Journal of Molecular Catalysis B:Enzymatic, 2014,103:72-78.
[54]	Austin HP, Allen MD, Donohoe BS, et al. Characterization and engineering of a plastic-degrading aromatic polyesterase[J]. Proc Natl Acad Sci USA, 2018,115(19):E4350-E4357. URL pmid: 29666242
[55]	Ma Y, Yao M, Li B, et al. Enhanced poly(ethylene terephthalate)hydrolase activity by protein engineering[J]. Engineering, 2018,4(6):888-893.
[56]	Liu B, He L, Wang L, et al. Protein crystallography and site-direct mutagenesis analysis of the poly(ethylene terephthalate)hydrolase PETase from Ideonella sakaiensis[J]. Chembiochem, 2018,19(14):1471-1475. doi: 10.1002/cbic.201800097 URL pmid: 29603535
[57]	Liu C, Shi C, Zhu S, et al. Structural and functional characterization of polyethylene terephthalate hydrolase from Ideonella sakaiensis[J]. Biochemical and Biophysical Research Communications, 2019,508(1):289-294. URL pmid: 30502092
[58]	Alves NM, Mano JF, Balaguer E, et al. Glass transition and structural relaxation in semi-crystalline poly(ethylene terephthalate):A DSC study[J]. Polymer, 2002,43(15):4111-4122. doi: 10.1016/S0032-3861(02)00236-7 URL
[59]	Chen CC, Dai L, Ma L, et al. Enzymatic degradation of plant biomass and synthetic polymers[J]. Nature Reviews Chemistry, 2020,4(3):114-126. doi: 10.1038/s41570-020-0163-6 URL
[60]	Kawai F, Oda M, Tamashiro T, et al. A novel Ca²+-activated, thermostabilized polyesterase capable of hydrolyzing polyethylene terephthalate from Saccharomonospora viridis AHK190[J]. Appl Microbiol Biotechnol, 2014,98(24):10053-10064. doi: 10.1007/s00253-014-5860-y URL
[61]	Son HF, Cho IJ, Joo S, et al. Rational protein engineering of thermo-stable PETase from Ideonella sakaiensis for highly efficient PET degradation[J]. ACS Catalysis, 2019,9(4):3519-3526. doi: 10.1021/acscatal.9b00568 URL
[62]	王泽方, 杨海涛, 王雪, 等. 细胞表面展示PET分解酶的重组毕赤酵母及构建与应用:中国, CN201610912589. 1[P]. 2016-10-20.
	Wang ZF, Yang HT, Wang X, et al. Recombinant Pichia pastoris for displaying PET lyase on cell surface and construction and application thereof:China, CN201610912589. 1[P]. 2016-10-20.
[63]	王泽方, 杨海涛, 王雪, 等. 细胞表面共展示PET分解酶和疏水蛋白的重组毕赤酵母:中国, CN201610912165. 5[P]. 2016-10-20.
	Wang ZF, Yang HT, Wang X, et al. Recombinant Pichia pastoris for displaying PET lyase and hydrophobin on cell surface:China, CN201610912165. 5[P]. 2016-10-20.
[64]	Furukawa M, Kawakami N, Oda K, et al. Acceleration of enzymatic degradation of poly(ethylene terephthalate)by surface coating with anionic surfactants[J]. ChemSusChem, 2018,11(23):4018-4025. doi: 10.1002/cssc.201802096 URL pmid: 30291679
[65]	Huang X, Cao L, Qin Z, et al. Tat-independent secretion of polyethylene terephthalate hydrolase PETase in Bacillus subtilis 168 mediated by its native signal peptide[J]. Journal of Agricultural and Food Chemistry, 2018,66(50):13217-13227. URL pmid: 30465427
[66]	Seo H, Kim S, Son HF, et al. Production of extracellular PETase from Ideonella sakaiensis using sec-dependent signal peptides in E. coli[J]. Biochemical and Biophysical Research Communications, 2019,508(1):250-255. URL pmid: 30477746
[67]	Moog D, Schmitt J, Senger J, et al. Using a marine microalga as a chassis for polyethylene terephthalate(PET)degradation[J]. Microb Cell Fact, 2019,18(1):171. URL pmid: 31601227
[68]	Chen ZZ, Wang YY, Cheng YY, et al. Efficient biodegradation of highly crystallized polyethylene terephthalate through cell surface display of bacterial PETase[J]. Science of the Total Environment, 2020,709:136138.
[69]	苏怡丹, 伍宁丰, 刘国安. 蛋白质定向进化技术的研究进展[J]. 生物技术通报, 2009(11):43-47.
	Su YD, Wu NF, Liu GA. Research on directed evolution of protein[J]. Biotechnology Bulletin, 2009(11):43-47.
[70]	付畅, 林海龙, 李海莹, 等. 蛋白质体外定向进化策略研究进展[J]. 生物技术通报, 2012(7):36-40.
	Fu C, Lin HL, Li HY, et al. Advances in study on directed molecular evolution strategy in vitro of protein[J]. Biotechnology Bulletin, 2012(7):36-40.
[71]	朱蕾蕾, 石利霞, 高松枫. 一种信号肽突变体及其应用:中国, CN202010025261. 4[P].
	Zhu LL, Shi LX, Gao SF. Signal peptide mutant and application thereof:China, CN202010025261. 4[P].
[72]	杨媛, 张剑. 微生物脂肪酶的性质及应用研究[J]. 中国洗涤用品工业, 2017(4):47-54.
	Yang Y, Zhang J. Study on properties and application of microbial lipase[J]. China Cleaning Industry, 2017(4):47-54.
[73]	Zimmermann W, Billig S. Enzymes for the biofunctionalization of poly(ethylene terephthalate)[J]. Adv Biochem Eng Biotechnol, 2011,125(1):97-120.
[74]	Fernandez, Lafuente R. Lipase from Thermomyces lanuginosus:Uses and prospects as an industrial biocatalyst[J]. Journal of Molecular Catalysis B:Enzymatic, 2010,62(3-4):197-212.
[75]	Noble ME, Cleasby A, Johnson LN, et al. The crystal structure of triacylglycerol lipase from Pseudomonas glumae reveals a partially redundant catalytic aspartate[J]. FEBS Letters, 1993,331(1-2):123-128. doi: 10.1016/0014-5793(93)80310-q URL pmid: 8405390
[76]	Brzozowski AM, Savage H, Verma CS, et al. Structural origins of the interfacial activation in Thermomyces(Humicola)lanuginosa lipase[J]. Biochemistry, 2000,39(49):15071-15082. doi: 10.1021/bi0013905 URL pmid: 11106485
[77]	Kawai F, Kawabata T, Oda M. Current knowledge on enzymatic PET degradation and its possible application to waste stream management and other fields[J]. Applied Microbiology and Biotechnology, 2019,103:4253-4268. URL pmid: 30957199
[78]	Palm GJ, Reisky L, Böttcher D, et al. Structure of the plastic-degrading Ideonella sakaiensis MHETase bound to a substrate[J]. Nature Communications, 2019,10(1):1717. doi: 10.1038/s41467-019-09326-3 URL pmid: 30979881

Source		Enzyme	PDB ID	Material	Temperature/℃	Time/h	Weight loss/%	References
Metagenomic	Leaf-branch compost	LCC	4EB0	lc PET film	70	48	95	[4-5]
Fungi	Humilica insolens	HiC	4OYY	lcPET film（7% crystallinity）	70	96	97±3	[6-7]
Fungi	Fusarium solani	FsC	1CEX	lcPET film（7% crystallinity）	40	96	5±1	[6,8]
bacterial	Thermobifida cellulosilytica	Thc_Cut1	5LUI	PET film（37% crystallinity）	50	72	-	[9-10]
	Thermobifida cellulosilytica	Thc_Cut2	5LUJ	PET film（37% crystallinity）	50	72	-	[9-10]
	Thermobifida fusca	TfH（BTA1）	-	PET film	65	48	12.9±1.2	[11]
	Thermobifida fusca KW3	TfCut2	4CG1	amorphous PET film	65	50	15.9±1.8	[12]

Source		Enzyme	PDB ID	Material	Temperature/℃	Time/h	Weight loss/%	References
Metagenomic	Leaf-branch compost	LCC	4EB0	lc PET film	70	48	95	[4-5]
Fungi	Humilica insolens	HiC	4OYY	lcPET film（7% crystallinity）	70	96	97±3	[6-7]
Fungi	Fusarium solani	FsC	1CEX	lcPET film（7% crystallinity）	40	96	5±1	[6,8]
bacterial	Thermobifida cellulosilytica	Thc_Cut1	5LUI	PET film（37% crystallinity）	50	72	-	[9-10]
	Thermobifida cellulosilytica	Thc_Cut2	5LUJ	PET film（37% crystallinity）	50	72	-	[9-10]
	Thermobifida fusca	TfH（BTA1）	-	PET film	65	48	12.9±1.2	[11]
	Thermobifida fusca KW3	TfCut2	4CG1	amorphous PET film	65	50	15.9±1.8	[12]

Source of Lipases	Type of PET	Effect on PET	References
Thermomyces lanuginosus	Fabric,fibers PET trimer	Hydrolysis increased hydrophilicity	[13-16]
Aspergillus sp.	Fabric, PET trimer	Hydrolysis increased hydrophilicity	[17]
Porcine（hog）pancreas	Fabric	Increased hydrophilicity changes in surface morphology	[18-20]
Burkholderia cepacia	Fabric, PET trimer	Hydrolysis increased hydrophilicity	[20]
Candida antarctica	Fabric,fibers	Hydrolysis increased hydrophilicity	[20-22]

Source of Lipases	Type of PET	Effect on PET	References
Thermomyces lanuginosus	Fabric,fibers PET trimer	Hydrolysis increased hydrophilicity	[13-16]
Aspergillus sp.	Fabric, PET trimer	Hydrolysis increased hydrophilicity	[17]
Porcine（hog）pancreas	Fabric	Increased hydrophilicity changes in surface morphology	[18-20]
Burkholderia cepacia	Fabric, PET trimer	Hydrolysis increased hydrophilicity	[20]
Candida antarctica	Fabric,fibers	Hydrolysis increased hydrophilicity	[20-22]