More efficient enzymatic cascade reactions by spatially confining enzymes via the SpyTag/SpyCatcher technology
نویسندگان
چکیده
Hydrocarbon synthesis from (waste)oils enabled by a cascade of lipase-catalysed hydrolysis and decarboxylase-catalysed decarboxylation has become an active area research en route to alternative, biobased fuels. However, Poor substrate transport efficiency is major issue causing low reaction rates. This study focused on protein self-assembly strategy based SpyTag/SpyCatcher overcome diffusion limitations. For this, two fusion proteins, TLL-Linker-SpyCatcher the lipase Thermomyces lanuginosus CvFAP-Linker-SpyTag fatty acid photodecarboxylase Chlorella variabilis were designed. A covalent multi-enzyme complex (TLL-CvFAP) was formed spontaneously each enzyme. The effects temperature, pH molar ratio self-assembled components assembly investigated. results showed that TLL-CvFAP reached about 60% after 12 h assembly, enzyme activity multienzyme increased 50% compared corresponding non-assembled enzymes. Under optimized conditions 10 mM soybean oil converted into 25 hydrocarbons, suggesting good potential biofuel synthesis.
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ژورنال
عنوان ژورنال: Molecular Catalysis
سال: 2022
ISSN: ['2522-5081', '2522-509X']
DOI: https://doi.org/10.1016/j.mcat.2022.112188