Microreactors derived from water-in-water pickering emulsions for Bacillus amyloliquefaciens proliferation and...

A microreactor derived from water-in-water (W/W) Pickering emulsion was developed to address the challenges of low yield and volatility in 4-methyl-5-thiazoleethanol (MTE) biosynthesis. The thiaminase II (TenA) from Bacillus amyloliquefaciens was engineered to create a key mutant (A82Y/Y113A/T163K/F169L), which was integrated into the microreactor system. The microreactor was constructed using methylcellulose (MD)/hydroxypropyl methylcellulose (HPMC) aqueous two-phase separation to encapsulate engineered probiotics and minimize volatile losses. An 8.86-fold increase in MTE production (from 281.72 to 2497.43 mg/L) was achieved by the engineered TenA through liquid fermentation. The microreactor further enhanced production to 2790.15 mg/L, representing a 14 % improvement over conventional methods. Volatility losses were effectively reduced, and enzyme-substrate interactions were optimized via interfacial barriers. A novel microreactor platform was established by combining synthetic biology with colloidal assembly, enabling efficient and sustainable MTE production. This strategy offers significant potential for industrial applications in the food and fragrance industries.