摘要
Pectate lyase(PEL)has broad industrial utility,but its application is limited by insufficient thermal stability and alkali tolerance.In this study,a novel PEL(hereafter,ScpB)from Phytophthora parasitica INRA-310 was identified for the first time through data-driven mining and bioinformatics.Homology modeling revealed that ScpB consists of two domains:a PEL superfamily-associated domain ScpB-Ⅰ and a catalytic domain ScpB-Ⅱ,which are connected by a short linker region.Based on truncated engineering and protein structure engineering strategies,ScpB-Ⅰ and ScpB-Ⅱ must cooperate to exhibit the PEL activity.Furthermore,ScpB was recruited for developing biochemical properties.The purified ScpB exhibited optimal activity at pH 10.0 and 55℃,aligning with textile biorefining conditions.Notably,in the presence of Ca^(2+)and Fe^(3+),a 215.97%and 182.08%,respectively,improvement in catalytic activity compared to the control group without metal ion.Finally,the bioscouring assay was carried out,ScpB exhibited a significant increase in the wetted fabric area to 5.371±0.382 cm^(2),which was 3.6 times that of the untreated sample(1.496 cm^(2)).Scanning electron microscopy confirmed effective pectin removal and smoother fibers.This study expands the PEL resource library and offers a promising enzyme for efficient,eco-friendly textile biorefining.
基金
funded by National Key R&D Program of China(2022YFC2105502)
the National Natural Science Foundation of China(32200061,22208124)
the Natural Science Foundation of Jiangsu Province of China(BK20210752)
the Postdoctoral Science Foun-dation of China(2022M721390,2023T160278)
Postgraduate Research&Practice Innovation Program of Jiangsu Province(KYCX24_4026).
