Structural and Functional Studies of <i>Aspergillus oryzae</i> Cutinase: Enhanced Thermostability and Hydrolytic Activity of Synthetic Ester and Polyester Degradation

Liu, Zhi‐Qiang; Gosser, Yuying; Baker, Peter J.; Ravee, Yaniv; Lu, Alvin; Alemu, Girum; Li, Huiguang; Butterfoss, Glenn L.; Kong, Xiang‐Peng; Gross, Richard A.; Montclare, Jin Kim

doi:10.1021/ja9046697

Cited by 121 publications

(112 citation statements)

References 47 publications

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“…AnCUT2 was shown to retain more than 60% of its relative activity after 1 h incubation at different pHs. Although it is a mesophilic enzyme, AnCUT2 exhibits remarkable stability after 1 h incubation at 40°C compared to G. cingulata cutinase [29] and F. solani [30]. Similar data have been previously observed in some fungal cutinases [30,31].…”

Section: Enzymatic Treatment Of Synthetic Polymerssupporting

confidence: 85%

“…Although it is a mesophilic enzyme, AnCUT2 exhibits remarkable stability after 1 h incubation at 40°C compared to G. cingulata cutinase [29] and F. solani [30]. Similar data have been previously observed in some fungal cutinases [30,31]. This thermostability observed may be due to serine (Ser) residues that represent the highest percentage (15.2%) of AnCUT2 composition.…”

Section: Enzymatic Treatment Of Synthetic Polymerssupporting

confidence: 79%

“…thermotolerance of AnCUT2 might be due to the presence of three disulphide bonds in the protein (Figure 1). A. oryzae cutinase is more stable than F. solani cutinase and this enhanced thermotolerance might be attributed to the presence of three bonds while the latter possesses only two [30].…”

Section: Enzymatic Treatment Of Synthetic Polymersmentioning

confidence: 98%

“…Activity towards short chain substrates may reflect structural features of the active site of the cutinase [30]. Fungal cutinases show different p-nitrophenyl substrate specificities; A. oryzae cutinase prefers pNPV [30], Sirococcus conigenus [41], T. harzianum [10] and F. solani [30] cutinases prefer pNPA (C2), while G. cingulata cutinase prefers pNPC (C8), followed by pNPM (C14), and pNPL (C12) [39].…”

Section: Conflict Of Interestsmentioning

confidence: 99%

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Expression and characterization of a cutinase (AnCUT2) from Aspergillus niger

et al. 2016

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Cutin hydrolase (EC 3.1.1.74), an extracellular polyesterase found in pollens, bacteria and fungi, is an efficient catalyst that exhibits hydrolytic activity on a variety of water-soluble esters, synthetic fibers, plastics and triglycerides. Thus, cutinase can be used in various applications such as ester synthesis, bio-scouring, food and detergent industries. Ancut2 is one of five genes encoding cutinases present in the Aspergillus niger ATCC 10574 genome. The cDNA of Ancut2 comprising of an open reading frame of 816 bp encoding a protein of 271 amino acid residues, was isolated and expressed in Pichia pastoris. The partially purified recombinant cutinase exhibited a molecular mass of approximately 40 kDa. The enzyme showed highest activity at 40°C with a preference for acidic pH (5.0-6.0). AnCUT2 showed hydrolytic activity towards various p-nitrophenyl esters with preference towards shorter chain esters such as p-nitrophenyl butyrate (C4). Scanning Electron Microscopy demonstrated that AnCUT2 was capable of modifying surfaces of synthetic polycaprolactone and polyethylene terephthalate plastics. The properties of this enzyme suggest that it may be applied in synthetic fiber modification and fruit processing industries.

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Section: Enzymatic Treatment Of Synthetic Polymerssupporting

confidence: 85%

Section: Enzymatic Treatment Of Synthetic Polymerssupporting

confidence: 79%

Section: Enzymatic Treatment Of Synthetic Polymersmentioning

confidence: 98%

Section: Conflict Of Interestsmentioning

confidence: 99%

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Expression and characterization of a cutinase (AnCUT2) from Aspergillus niger

et al. 2016

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“…To date, the threedimensional (3D) structure has been reported for three cutinases, from Fusarium solani (FsC), 1 Glomerella cingulata (GcC), 2,3 and Aspergillus oryzae (AoC). 4 These enzymes are members of the a/b hydrolase superfamily with a central parallel bsheet and surrounding a-helices, and active site consisting of a catalytic triad of a serine, histidine, and aspartic acid. Cutinases show no interfacial activation and are, therefore, placed as intermediates between lipases and esterases rather than being categorized as classical lipases.…”

Section: Introductionmentioning

confidence: 99%

Thermodynamic and structural investigation of the specific SDS binding of humicola insolens cutinase

et al. 2014

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The interaction of lipolytic enzymes with anionic surfactants is of great interest with respect to industrially produced detergents. Here, we report the interaction of cutinase from the thermophilic fungus Humicola insolens with the anionic surfactant SDS, and show the enzyme specifically binds a single SDS molecule under nondenaturing concentrations. Protein interaction with SDS was investigated by NMR, ITC and molecular dynamics simulations. The NMR resonances of the protein were assigned, with large stretches of the protein molecule not showing any detectable resonances. SDS is shown to specifically interact with the loops surrounding the catalytic triad with medium affinity (K a % 10 5 M 21 ). The mode of binding is closely similar to that seen previously for binding of amphiphilic molecules and substrate analogues to cutinases, and hence SDS acts as a substrate mimic. In addition, the structure of the enzyme has been solved by X-ray crystallography in its apo form and after cocrystallization with diethyl p-nitrophenyl phosphate (DNPP) leading to a complex with monoethylphosphate (MEP) esterified to the catalytically active serine. TheAbbreviations: AA, all-atom; AoC, Aspergillus oryzae cutinase; AOT, sodium bis(2-ethylhexyl) sulfosuccinate; cmc, critical micelle concentration; DEP, diethylphosphate; DNPP, diethyl p-nitrophenyl phosphate; EDTA, ethylene diamine tetraacetate; FsC, Fusarium solani cutinase; GcC, Glomerella cingulata cutinase; HiC, Humicola insolens cutinase; HSQC, heteronuclear singlequantum coherence; IPTG, isopropyl b-D-1-thiogalactopyranoside; ITC, isothermal titration calorimetry; MD, molecular dynamics; MEP, monoethylphosphate; MWCO, molecular weight cut-off; PAGE, polyacrylamide gel electrophoresis; RMSD, root mean square deviation; RMSF, root mean square fluctuation; TES, N-[tris(hydroxymethyl)methyl]-2-aminoethanesulfonic acid; SANS, small-angle neutron scattering; SDS, sodium dodecyl sulfate.Additional Supporting Information may be found in the online version of this article.An interactive view is available in the electronic version of the article. enzyme has the same fold as reported for other cutinases but, unexpectedly, esterification of the active site serine is accompanied by the ethylation of the active site histidine which flips out from its usual position in the triad.

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2023

Conformational Analysis of Polymers

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Structural and Functional Studies of Aspergillus oryzae Cutinase: Enhanced Thermostability and Hydrolytic Activity of Synthetic Ester and Polyester Degradation

Cited by 121 publications

References 47 publications

Expression and characterization of a cutinase (AnCUT2) from Aspergillus niger

Expression and characterization of a cutinase (AnCUT2) from Aspergillus niger

Thermodynamic and structural investigation of the specific SDS binding of humicola insolens cutinase

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