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Protein expression and purification, 2021-11, Vol.187, p.105933-105933, Article 105933
2021
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Autor(en) / Beteiligte
Titel
Recombinant expression and surface display of a zearalenone lactonohydrolase from Trichoderma aggressivum in Escherichia coli
Ist Teil von
  • Protein expression and purification, 2021-11, Vol.187, p.105933-105933, Article 105933
Ort / Verlag
United States: Elsevier Inc
Erscheinungsjahr
2021
Quelle
MEDLINE
Beschreibungen/Notizen
  • Zearalenone (ZEN), one of the most dangerous mycotoxins, causes enormous economic losses in the food and feed industries. To solve the problem of ZEN pollution, ZEN detoxifying enzymes are in emergent need. In this study, a zearalenone lactonohydrolase from Trichoderma aggressivum, denoted as ZHD-P, was heterologously expressed and characterized. The intracellular ZHD-P from E. coli BL21(DE3) exhibited high activity for ZEN degradation (191.94 U/mg), with the optimal temperature and pH of 45 °C and 7.5–9.0, respectively. With excellent temperature stability, the intracellular ZHD-P retained 100% activity when it was incubated at 25–40 °C for 1 h. Furthermore, we firstly constructed an E. coli cell surface display system for ZHD-P. The surface-displayed ZHD-P exhibited high activity against ZEN and showed optimal activity at 40 °C and pH 9.0. With superior pH stability, the surface-displayed ZHD-P retained 80% activity when it was incubated at pH 5.0–11.0 for 12 h. Interestingly, the metal ions tolerance of the surface-displayed ZHD-P was better than the intracellular form. Additionally, the surface-displayed ZHD-P could be reused four times with the residual enzyme activity of more than 50%. The biotoxicity assessment using P. phosphoreum T3 indicated that ZEN could be degraded into hypotoxic products by the intracellular or surface-displayed ZHD-P. ZHD-P could be feasible for ZEN detoxification. •A zearalenone lactonohydrolase ZHD-P, from Trichoderma aggressivum, was expressed in E. coli BL21(DE3) and successfully purified. Zearalenone degradation activity of the intracellular ZHD-P achieved 191.94 U/mg.•An E. coli cell surface display system for ZHD-P was constructed. The surface-displayed ZHD-P exhibited high activity against zearalenone.•Characterizations of the intracellular and surface-displayed ZHD-P were performed by measuring the optimal temperature, pH and the effects of metal ions.

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