Characterization of a Novel Thermostable and Alkaliphilic β-Mannanase for Gel-Breaking in Guar Gum Fracturing Fluids

The development of robust and efficient β-mannanases is key to advancing environmentally friendly industrial processes, such as guar gum fracturing fluid gel-breaking. Here, we report the identification and characterization of MG4, a novel thermotolerant and alkaliphilic β-mannanase mined from the Earth’s Microbiome database. The recombinant enzyme has a molecular weight of 63 kDa. MG4 displayed maximum activity at 65 °C and pH 9.0, and exhibited remarkable stability across a broad pH range (7.0–10.0). It retained over 80% of its activity after incubation at 50 °C for 1 h, and its activity was enhanced more than 40% by Mg2+ or Ca2+. Moreover, MG4 (20 mg/L) reduced the viscosity of guar gum fracturing fluid to <5 m·PaS within 30 min, outperforming ammonium persulfate (APS, 500 mg/L) which required 1 h, and produced 64.5% less insoluble residue. TEM imaging directly visualized the disruption of the guar gum polymer network by MG4, explaining its efficacy and suggesting reduced formation damage risk compared to chemical breakers. This work characterizes a highly promising biocatalyst whose thermostability, alkaliphily, efficient gel-breaking, low residue yield, and minimal formation damage potential position it as a superior, eco-friendly alternative for petroleum industry applications.