Functional characterisation of a metagenome derived family VIII esterase with a deacetylation activity on β-lactam antibiotics

Mokoena, Nobalanda B.; Mathiba, Kgama; Tsekoa, Tsepo L.; Steenkamp, Paul A.; Rashamuse, Konanani

doi:10.1016/j.bbrc.2013.06.076

Cited by 26 publications

(25 citation statements)

References 22 publications

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“…No enzyme activity was noticed toward p -nitrophenyl phosphate ( p -NP). Similarly, EstU1, Est22, EstM-N1 or EstC showed substrate preference for p -nitrophenyl butyrate (C4), while no enzyme activities of these enzymes was observed for p -nitrophenyl esters with longer chains (from C12 to C18)12131415. When diverse naphthyl derivatives were used as substrates, the highest activities were obtained with 2-naphthyl acetate (2-NA), followed by 1-naphthyl acetate (1-NA), and 1-naphthyl butyrate (1-NB).…”

Section: Resultsmentioning

confidence: 99%

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Biochemical and Structural Analysis of a Novel Esterase from Caulobacter crescentus related to Penicillin-Binding Protein (PBP)

Ryu

Ngo

Yoo

et al. 2016

Sci Rep

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Considering that the prevalence of antibiotic-resistant pathogenic bacteria is largely increasing, a thorough understanding of penicillin-binding proteins (PBPs) is of great importance and crucial significance because this enzyme family is a main target of β-lactam-based antibiotics. In this work, combining biochemical and structural analysis, we present new findings that provide novel insights into PBPs. Here, a novel PBP homologue (CcEstA) from Caulobacter crescentus CB15 was characterized using native-PAGE, mass spectrometry, gel filtration, CD spectroscopy, fluorescence, reaction kinetics, and enzyme assays toward various substrates including nitrocefin. Furthermore, the crystal structure of CcEstA was determined at a 1.9 Å resolution. Structural analyses showed that CcEstA has two domains: a large α/β domain and a small α-helix domain. A nucleophilic serine (Ser68) residue is located in a hydrophobic groove between the two domains along with other catalytic residues (Lys71 and Try157). Two large flexible loops (UL and LL) of CcEstA are proposed to be involved in the binding of incoming substrates. In conclusion, CcEstA could be described as a paralog of the group that contains PBPs and β-lactamases. Therefore, this study could provide new structural and functional insights into the understanding this protein family.

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Section: Resultsmentioning

confidence: 99%

“…These include EstU112, Est2213, EstM-N114, and EstC15. In a previous study, identification and preliminary X-Ray diffraction analysis of a novel PBP homolog (CcEstA, CCNA_00255) in Caulobacter crescentus CB15 were reported16.…”

mentioning

confidence: 99%

Biochemical and Structural Analysis of a Novel Esterase from Caulobacter crescentus related to Penicillin-Binding Protein (PBP)

Ryu

Ngo

Yoo

et al. 2016

Sci Rep

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“…4IVK) (39). Promiscuous ␤-lactamase activity has been demonstrated for some members of this family, such as the metagenome-derived carboxylesterases EstU1 (40) and Est22 (41), with first-generation cephalosporin-based derivatives. EstG4 clustered in family II of lipolytic enzymes, which show the conserved motif GDSL containing the active site serine.…”

Section: Resultsmentioning

confidence: 99%

Discovery of Polyesterases from Moss-Associated Microorganisms

Müller

Perz

Provasnek

et al. 2017

Appl Environ Microbiol

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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.

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“…The majority either lack the activity or show negligible activity, despite the high sequence identity to the β-lactamases, while others have been described as exhibiting "promiscuous β-lactamase activity" [18,21,41,47,48,49]. Consequently, it has been suggested that these esterases have evolved from the class C β-lactamases, where some have maintained this remnant activity, while others have lost the capability due to steric interference resulting from structural evolution [18,41,50].…”

Section: Discussionmentioning

confidence: 99%

An unusual feruloyl esterase belonging to family VIII esterases and displaying a broad substrate range

Ohlhoff

Kirby

Zyl

et al. 2015

Journal of Molecular Catalysis B: Enzymatic

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Functional characterisation of a metagenome derived family VIII esterase with a deacetylation activity on β-lactam antibiotics

Cited by 26 publications

References 22 publications

Biochemical and Structural Analysis of a Novel Esterase from Caulobacter crescentus related to Penicillin-Binding Protein (PBP)

Biochemical and Structural Analysis of a Novel Esterase from Caulobacter crescentus related to Penicillin-Binding Protein (PBP)

Discovery of Polyesterases from Moss-Associated Microorganisms

An unusual feruloyl esterase belonging to family VIII esterases and displaying a broad substrate range

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