Bioremediation of pulp and paper industry (PPI) effluent is hindered due to biorefractory lignin. Herein, we demonstrate a lignin-specific designer biosurfactant with synergistic binding sites for enhanced lignin removal from PPI effluent. The histidine rich-cationic lipoprotein biosurfactant (HR-CLB) was synthesized by Bacillus tropicus using tanning industry solid waste, animal fleshing by de novo substrate-dependent synthesis pathway. Interestingly, the HR-CLB anchored functionalized carbon (HR-CLBAFC) demonstrated a high lignin sequestration capacity of 93.2 mg/g HR-CLBAFC at optimized time, 60 min; pH, 5; temperature, 45 °C; and mass of HR-CLBAFC, 1.0 g. The sequestration was confirmed by HR-SEM, UV–Vis., FT-IR, and WCA analyses. The isotherm studies revealed the involvement of Freundlich isotherm (regression coefficient, R2: 0.988) in regulating lignin sequestration onto HR-CLBAFC with a Freundlich constant (Kf) of 16.46 ((mg/g)(L/mg)1/n). Moreover, the kinetics studies divulged the contribution of the pseudo-second-order kinetic model (R2: 0.992) in regulating the dynamic mechanism of lignin sequestration onto HR-CLBAFC with a rate constant (k2) of 0.00022 g/mg min. Additionally, the thermodynamics studies discovered a positive Gibbs free energy (ΔG: +170 kJ/mol) and entropy (ΔS°: +130 kJ/mol), indicating the involvement of chemical interaction during the sequestration. Furthermore, the mechanistic study confirmed the role of an incomplete valence shell of nitrogen in histidine centers of HR-CLB in regulating electrostatic interaction with lignin molecules during the sequestration process. Subsequently, the HR-CLBAFC applied for lignin sequestration from the real-time PPI effluent demonstrated an outstanding sequestration efficiency (>99.4%), confirming the unequivocal potential of the HR-CLBAFC matrix in lignin sequestration from real-time PPI effluent.
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