Converting Bacillus subtilis 168 to a Synthetic Methylotroph by Combinatorial Metabolic Regulation Strategies

• DOI: 10.1021/acs.jafc.4c09781
• Publication Date: 2025-02-12T16:49:51Z
• Authors: Qingfang Meng; Dexin Wang; Xiaoping Fu; Weitao Geng; Hongchen Zheng; Wenqin Bai
• Keywords: Methylotroph; Bacillaceae; Combinatorial synthesis

Methanol, which can come from methane or carbon dioxide, is a valuable renewable one-carbon (C1) feedstock for the production of biofuels and food chemicals. A new method was developed to create multienzyme complex by combining methanol dehydrogenase (Mdh), 3-hexulose-6-phosphate synthase (Hps), 6-phospho-3-hexuloseisomerase (Phi) in equal parts using SpyTag/Catcher DogTag/Catcher systems. This self-assembly multiple enzymes improves conversion fructose-6-phosphate (F6P) used engineer synthetic methylotroph B. subtilis 168 that could efficiently utilize methanol. Various metabolic regulations related key pathways were tested integrated boost consumption this engineered strain. The final strain, SM6, consume 3.87 g/L methanol, marking highest level coutilization with xylose date. strategies employed research optimized distribution flow formaldehyde xylose, offering insights future studies on methylotrophs.