Substrate specificities of cutinases on aliphatic–aromatic polyesters and on their model substrates

Perz, Veronika; Bleymaier, Klaus; Sinkel, Carsten; Kueper, Ulf; Bonnekessel, Melanie; Ribitsch, Doris; Guebitz, Georg M.

doi:10.1016/j.nbt.2015.11.004

Cited by 66 publications

(90 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Enzymes that exhibit the ability to hydrolyze PBAT were assumed to be found within the family of extracellular serine hydrolases since members of this enzyme class like lipases (EC 3.1.1.3), carboxylesterases (EC 3.1.1.1) and cutinases (3.1.1.74) have been reported to hydrolyze (synthetic) polymers. Lipases (Marten et al, ) and cutinases (Herrero Acero et al, ; Perz et al, ; Ribitsch et al, ) have already been described to hydrolyze synthetic polymers like PET or PBAT.…”

Section: Resultsmentioning

confidence: 99%

Hydrolysis of synthetic polyesters by Clostridium botulinum esterases

Perz

Baumschlager

Bleymaier

et al. 2015

Biotech & Bioengineering

Self Cite

View full text Add to dashboard Cite

Two novel esterases from the anaerobe Clostridium botulinum ATCC 3502 (Cbotu_EstA and Cbotu_EstB) were expressed in Escherichia coli BL21-Gold(DE3) and were found to hydrolyze the polyester poly(butylene adipate-co-butylene terephthalate) (PBAT). The active site residues (triad Ser, Asp, His) are present in both enzymes at the same location only with some amino acid variations near the active site at the surrounding of aspartate. Yet, Cbotu_EstA showed higher kcat values on para-nitrophenyl butyrate and para-nitrophenyl acetate and was considerably more active (sixfold) on PBAT. The entrance to the active site of the modeled Cbotu_EstB appears more narrowed compared to the crystal structure of Cbotu_EstA and the N-terminus is shorter which could explain its lower activity on PBAT. The Cbotu_EstA crystal structure consists of two regions that may act as movable cap domains and a zinc metal binding site.

show abstract

Section: Resultsmentioning

confidence: 99%

Hydrolysis of synthetic polyesters by Clostridium botulinum esterases

Perz

Baumschlager

Bleymaier

et al. 2015

Biotech & Bioengineering

Self Cite

View full text Add to dashboard Cite

show abstract

“…Generally, higher concentrations of Ta versus BTaB and BTa indicate more efficient enzymatic hydrolysis of the aromatic ester bond, i.e., the Ta-B bond (12). The highest concentrations of released Ta (up to 35 M) were measured with milled PBAT and EstB3, EstB11, and EstC7 (Fig.…”

Section: Resultsmentioning

confidence: 89%

“…Similar differences in hydrolytic cleavage patterns for aliphatic-aromatic polyesters between enzymes were reported previously, for example, for the cutinases from Humicola insolens (HiC) and Thermobifida cellulosilytica (Thc_Cut1). The cutinase Thc_ Cut1 shows greater specificity for the hydrolysis of aromatic ester bonds (Ta-B) than does the cutinase HiC (12). Moreover, the polyesterase activities of the novel enzymes EstB3 and EstC7 with milled PBAT were significantly higher than the activities of the recently presented esterases from the anaerobic bacteria Clostridium hathewayi (Chath_Est1) (21) and Clostridium botulinum (Cbotu_Est and Cbotu_EstB) (20).…”

Section: Fig 6 Hydrolysis Of Milled Pbat (A) and Pbat Foil (B) By Purmentioning

confidence: 95%

“…The hydrolysis products Ta, Ada, benzoic acid (Ba), BTa, and BTaB were quantified by means of RP-HPLC using a Dionex system (Dionex, USA), consisting of an UltiMate 3000 pump, an ASI-100 automated sample injector, an UltiMate 3000 column compartment, and a UVD 340U photodiode array detector, in combination with a XTerra RP 18 column (3.5 m; 3.0 mm by 150 mm) with a precolumn (Waters Corp., USA). Separation of the hydrolysis products was performed as described previously (12). Signals from blank and control reactions were subtracted from the sample signals in order to correct the data for background activity (autohydrolysis).…”

Section: Methodsmentioning

confidence: 99%

“…Novel biocatalysts capable of hydrolyzing polyesters could allow recycling of valuable building blocks. Microbial enzymes from various sources (e.g., compost) that can degrade or modify natural and synthetic polyesters have been studied previously (11,12). Esterases and lipases not only are prevalent in most natural ecosystems and microorganisms but also are some of the most widely used biocatalysts in the chemical industry (13).…”

mentioning

confidence: 99%

See 2 more Smart Citations

Discovery of Polyesterases from Moss-Associated Microorganisms

Müller

Perz

Provasnek

et al. 2017

Appl Environ Microbiol

Self Cite

View full text Add to dashboard Cite

The growing pollution of the environment with plastic debris is a global threat which urgently requires biotechnological solutions. Enzymatic recycling not only prevents pollution but also would allow recovery of valuable building blocks. Therefore, we explored the existence of microbial polyesterases in microbial communities associated with the Sphagnum magellanicum moss, a key species within unexploited bog ecosystems. This resulted in the identification of six novel esterases, which were isolated, cloned, and heterologously expressed in Escherichia coli. The esterases were found to hydrolyze the copolyester poly(butylene adipate-co-butylene terephthalate) (PBAT) and the oligomeric model substrate bis[4-(benzoyloxy)butyl] terephthalate (BaBTaBBa). Two promising polyesterase candidates, EstB3 and EstC7, which clustered in family VIII of bacterial lipolytic enzymes, were purified and characterized using the soluble esterase substrate p-nitrophenyl butyrate (K m values of 46.5 and 3.4 M, temperature optima of 48°C and 50°C, and pH optima of 7.0 and 8.5, respectively). In particular, EstC7 showed outstanding activity and a strong preference for hydrolysis of the aromatic ester bond in PBAT. Our study highlights the potential of plant-associated microbiomes from extreme natural ecosystems as a source for novel hydrolytic enzymes hydrolyzing polymeric compounds.IMPORTANCE In this study, we describe the discovery and analysis of new enzymes from microbial communities associated with plants (moss). The recovered enzymes show the ability to hydrolyze not only common esterase substrates but also the synthetic polyester poly(butylene adipate-co-butylene terephthalate), which is a common material employed in biodegradable plastics. The widespread use of such synthetic polyesters in industry and society requires the development of new sustainable technological solutions for their recycling. The discovered enzymes have the potential to be used as catalysts for selective recovery of valuable building blocks from this material.

show abstract

Insights into polyester plastic biodegradation by carboxyl ester hydrolases

Gricajeva

Nadda

Gudiukaitė

2021

J of Chemical Tech & Biotech

View full text Add to dashboard Cite

Environmental pollution caused by polyesters has become a major ecological safety concern that needs to be managed urgently. One way to resolve this problem is giving the spotlight to current emerging research of microbial biocatalysts. During the last two decades many researchers have reported the ability to break down and modify natural and synthetic polyesters using different microbial carboxyl ester hydrolases (lipases, esterases, cutinases, PETases, etc.) also called polyesterases, and contribution of these enzymes towards the reduction of plastic levels in the future. Continuous search of such lipolytic biocatalysts and their improvement via protein engineering strategies results in beneficial findings making the use of polyesterases in the biodegradation of plastics increasingly more realistic. The present review provides a comprehensive insight into the structural properties enabling microbial lipolytic-type carboxyl ester hydrolases to effectively catalyze the cleavage of ester linkages in different polyester plastics. Moreover, the management of extensively used polyester plastics using different lipolytic enzymes as an innovative eco-friendly solution is presented in this report. Furthermore, improvement of polyesterases via protein engineering for the development of more effective and suitable polyester-degrading lipolytic biocatalysts is summarized in this review as well.

show abstract

Substrate specificities of cutinases on aliphatic–aromatic polyesters and on their model substrates

Cited by 66 publications

References 33 publications

Hydrolysis of synthetic polyesters by Clostridium botulinum esterases

Hydrolysis of synthetic polyesters by Clostridium botulinum esterases

Discovery of Polyesterases from Moss-Associated Microorganisms

Insights into polyester plastic biodegradation by carboxyl ester hydrolases

Contact Info

Product

Resources

About