Son, Hyeoncheol Francis Cho, In Jin Joo, Seongjoon Seo, Hogyun Sagong, Hye-Young Young Choi, So Lee, Sang Yup Kim, Kyung-Jin Rational Protein Engineering of Thermo-Stable PETase from Ideonella sakaiensis for Highly Efficient PET Degradation Widespread utilization of polyethylene terephthalate (PET) has caused a variety of environmental and health problems; thus, the enzymatic degradation of PET can be a promising solution. Although PETase from Ideonalla sakaiensis (<i>Is</i>PETase) has been reported to have the highest PET degradation activity under mild conditions of all PET-degrading enzymes reported to date, its low thermal stability limits its ability for efficient and practical enzymatic degradation of PET. Using the structural information on <i>Is</i>PETase, we developed a rational protein engineering strategy using several <i>Is</i>PETase variants that were screened for high thermal stability to improve PET degradation activity. In particular, the <i>Is</i>PETase<sup>S121E/D186H/R280A</sup> variant, which was designed to have a stabilized β6-β7 connecting loop and extended subsite IIc, had a <i>T</i><sub>m</sub> value that was increased by 8.81 °C and PET degradation activity was enhanced by 14-fold at 40 °C in comparison with <i>Is</i>PETase<sup>WT</sup>. The designed structural modifications were further verified through structure determination of the variants, and high thermal stability was further confirmed by a heat-inactivation experiment. The proposed strategy and developed variants represent an important advancement for achieving the complete biodegradation of PET under mild conditions. stability limits;Thermo-Stable PETase;protein engineering strategy;subsite IIc;PET degradation activity;Rational Protein Engineering;PETase S 121E variant;PETase WT;T m value;Efficient PET Degradation;Ideonalla sakaiensis;PETase variants;PET-degrading enzymes;Ideonella sakaiensis;structure determination;heat-inactivation experiment;health problems;polyethylene terephthalate 2019-03-11
    https://acs.figshare.com/articles/dataset/Rational_Protein_Engineering_of_Thermo-Stable_PETase_from_Ideonella_sakaiensis_for_Highly_Efficient_PET_Degradation/7873895
10.1021/acscatal.9b00568.s002