Unraveling Structural and Biochemical Insights into a Novel Thermo-Alkaline Pectate Lyase from Caldicellulosiruptor bescii for Sustainable Fabric Bioscouring

Pectate lyase (PL) holds significant potential for applications in various industries. However, the existing PL was unable to adapt to thermo-alkaline industrial environments. In this work, a PL from Caldicellulosiruptor bescii DSM 6725 (hereafter, CbPelD) was studied to disclose its structural and biochemical properties. CbPelD consisted of a putative signal peptide sequence, a short linker region, and a catalytic domain (PelD-II), along with an exoacting β-fructofuranosidase domain (PelD-I). In molecular dynamics simulation, PelD-II demonstrated significantly higher activity and thermal stability compared to other truncated enzymes of CbPelD. The optimal pH and temperature for PelD-II were 10.0 and 60 °C, respectively, with the specific activity increasing by 164-223% in the presence of Ca2+ or Ni2+. The exotype PelD-II exhibited efficient degradation of polygalacturonic acid (PGA) into unsaturated galacturonic acid (uG1), digalacturonic acid (uG2), and trigalacturonic acid (uG3). In the bioscouring assay, PelD-II exhibited a significant increase in the wetted fabric area to 6.072 ± 0.684 cm2, which was about 7.9 times higher than untreated fabric. Scanning electron microscopy further revealed that treatment with PelD-II resulted in a smoother fiber surface, providing direct visual confirmation of the enzyme's action. These results highlight the potential of PelD-II for applications in the textile industries.