Permeabilized TreS-Expressing Bacillus subtilis Cells Decorated with Glucose Isomerase and a Shell of ZIF-8 as a Reusable Biocatalyst for the Coproduction of Trehalose and Fructose.

Metal-organic frameworks (MOFs) are a class of porous materials with versatile properties. In this study, ZIF-8 was employed to establish a two-enzyme system by encapsulating permeabilized Bacillus subtilis cells coated with glucose isomerase. B. subtilis was constructed by introducing the shuttle plasmid PMA5 associated with the overexpression of trehalose synthase. Using this two-enzyme system, trehalose was produced by trehalose synthase and the byproduct glucose was converted to fructose with the help of glucose isomerase. The decrease in glucose production not only relieved the inhibition of the entire reaction chain but also increased the final yield of trehalose. The highest trehalose production rate reached 67.7% and remained above 50% after 20 batches. In addition, the toxicity of the ZIF-8 coating for B. subtilis was investigated by fluorescence microscopy and was found to be negligible. By simulating an extreme environment, the ZIF-8 coating was demonstrated to have a protective effect on the cells and enzymes. This study provides a theoretical basis for the application of MOFs in the immobilization of microorganisms and enzymes.