Programmable Polyproteams of Tyrosine Ammonia Lyases as Cross-Linked Enzymes for Synthesizing <i>p</i>-Coumaric Acid
2022
Mingyu Jia | Zhiyuan Luo | Haomin Chen | Bianqin Ma | Li Qiao | Qinjie Xiao | Pengfei Zhang | Anming Wang
Ideal immobilization with enhanced biocatalyst activity and thermostability enables natural enzymes to serve as a powerful tool to yield synthetically useful chemicals in industry. Such an enzymatic method strategy becomes easier and more convenient with the use of genetic and protein engineering. Here, we developed a covalent programmable polyproteam of tyrosine ammonia lyases (<i>TAL-CLEs</i>) by fusing SpyTag and SpyCatcher peptides into the <i>N</i>-terminal and <i>C</i>-terminal of the TAL, respectively. The resulting circular enzymes were clear after the spontaneous isopeptide bonds formed between the SpyTag and SpyCatcher. Furthermore, the catalytic performance of the <i>TAL-CLEs</i> was measured via a synthesis sample of <i>p</i>-Coumaric acid. Our <i>TAL-CLEs</i> showed excellent catalytic efficiency, with 98.31 ± 1.14% yield of the target product—which is 4.15 ± 0.08 times higher than that of traditional glutaraldehyde-mediated enzyme aggregates. They also showed over four times as much enzyme-activity as wild-type TAL does and demonstrated good reusability, and so may become a good candidate for industrial enzymes.
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