Characterization of AnCUT3, a plastic-degrading paucimannose cutinase from Aspergillus niger expressed in Pichia pastoris

Altammar, Khadijah A.; Ling, Jonathan Guyang; Al-Bajalan, Hussein M.; Chin, Iuan-Sheau; Mackeen, Mukram Mohamed; Mahadi, Nor Muhammad; Murad, Abdul Munir Abdul; Bakar, Farah Diba Abu

doi:10.1016/j.ijbiomac.2022.10.022

Cited by 8 publications

(8 citation statements)

References 89 publications

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“…PMSF is a serin-protease inhibitor; cutinases also have a catalytic serin that PMSF can inhibit. As expected, PMSF has an inhibitory effect in ANCUT1, although this was not detrimental; this has also been observed in other cutinases (Altammar et al 2022 ).…”

Section: Resultssupporting

confidence: 82%

ANCUT1, a novel thermoalkaline cutinase from Aspergillus nidulans and its application on hydroxycinnamic acids lipophilization

Peña-Montes,

Bermúdez-García,

Castro-Ochoa

et al. 2024

Biotechnol Lett

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One of the four cutinases encoded in the Aspergillus nidulans genome, ANCUT1, is described here. Culture conditions were evaluated, and it was found that this enzyme is produced only when cutin is present in the culture medium, unlike the previously described ANCUT2, with which it shares 62% amino acid identity. The differences between them include the fact that ANCUT1 is a smaller enzyme, with experimental molecular weight and pI values of 22 kDa and 6, respectively. It shows maximum activity at pH 9 and 60 °C under assayed conditions and retains more than 60% of activity after incubation for 1 h at 60 °C in a wide range of pH values (6–10) after incubations of 1 or 3 h. It has a higher activity towards medium-chain esters and can modify long-chain length hydroxylated fatty acids constituting cutin. Its substrate specificity properties allow the lipophilization of alkyl coumarates, valuable antioxidants and its thermoalkaline behavior, which competes favorably with other fungal cutinases, suggests it may be useful in many more applications.

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Section: Resultssupporting

confidence: 82%

ANCUT1, a novel thermoalkaline cutinase from Aspergillus nidulans and its application on hydroxycinnamic acids lipophilization

Peña-Montes,

Bermúdez-García,

Castro-Ochoa

et al. 2024

Biotechnol Lett

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“…The optimal pH and temperature for enzyme activity were determined using para-nitrophenyl dodecanoate as substrate according to the method of Altammar and colleagues 49 . Within a pH range from pH 3 to pH 8, for one hour at 30 °C, pH 7.0 appeared as the optimal pH.…”

Section: Resultsmentioning

confidence: 99%

“…11 . Plasmid loss was achieved by restreaking strains for several generations on SFM agar without antibiotic pressure at 37 °C and checking for the loss of apramycin resistance 49 , 50 .…”

Section: Methodsmentioning

confidence: 99%

“…The concentration of enzyme was estimated using the Bradford method (Bio-Rad, Bradford 1x Dye Reagent 5000205). The esterase/cutinase activity was tested using para-nitrophenyl dodecanoate (Sigma-Aldrich, 61716) The protocol followed was based on the protocol of Altammar and colleagues with minor adjustments 49 . For the optimal pH test, 50 mM citrate buffers ranging from pH 3 to pH 7 were used, and for pH 7.5 and 8 50 mM Tris-HCl buffer was used.…”

Section: Methodsmentioning

confidence: 99%

“…For the optimal pH test, 50 mM citrate buffers ranging from pH 3 to pH 7 were used, and for pH 7.5 and 8 50 mM Tris-HCl buffer was used. The incubation step of 10 min was prolonged to 1 h. The reaction was terminated using 0.1 M sodium carbonate 49 …”

Section: Methodsmentioning

confidence: 99%

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Polyester degradation by soil bacteria: identification of conserved BHETase enzymes in Streptomyces

Verschoor,

Croese,

Lakemeier

et al. 2024

Commun Biol

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The rising use of plastic results in an appalling amount of waste which is scattered into the environment. One of these plastics is PET which is mainly used for bottles. We have identified and characterized an esterase from Streptomyces, annotated as LipA, which can efficiently degrade the PET-derived oligomer BHET. The Streptomyces coelicolorScLipA enzyme exhibits varying sequence similarity to several BHETase/PETase enzymes, including IsPETase, TfCut2, LCC, PET40 and PET46. Of 96 Streptomyces strains, 18% were able to degrade BHET via one of three variants of LipA, named ScLipA, S2LipA and S92LipA. SclipA was deleted from S. coelicolor resulting in reduced BHET degradation. Overexpression of all LipA variants significantly enhanced BHET degradation. All variants were expressed in E. coli for purification and biochemical analysis. The optimum conditions were determined as pH 7 and 25 °C for all variants. The activity on BHET and amorphous PET film was investigated. S2LipA efficiently degraded BHET and caused roughening and indents on the surface of PET films, comparable to the activity of previously described TfCut2 under the same conditions. The abundance of the S2LipA variant in Streptomyces suggests an environmental advantage towards the degradation of more polar substrates including these polluting plastics.

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Commercialization potential of PET (polyethylene terephthalate) recycled nanomaterials: A review on validation parameters

Kumar,

Sharma,

Umesh

et al. 2024

Chemosphere

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Characterization of AnCUT3, a plastic-degrading paucimannose cutinase from Aspergillus niger expressed in Pichia pastoris

Cited by 8 publications

References 89 publications

ANCUT1, a novel thermoalkaline cutinase from Aspergillus nidulans and its application on hydroxycinnamic acids lipophilization

ANCUT1, a novel thermoalkaline cutinase from Aspergillus nidulans and its application on hydroxycinnamic acids lipophilization

Polyester degradation by soil bacteria: identification of conserved BHETase enzymes in Streptomyces

Commercialization potential of PET (polyethylene terephthalate) recycled nanomaterials: A review on validation parameters

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