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Target of this project is to employ a directed evolution approach aimed at improving cystathionine β-lyase (CBL) catalytic activity toward cysteine-S-conjugates. Directed evolution is a powerful tool for reshaping the functional and structural features of proteins: through iterative cycles of mutation, selection and amplification, important traits of biocatalysts can be optimized for industrial application. We will perform directed evolution of CBL by using error-prone PCR to create a library of variants that will be subsequently screened for improved activity toward cysteine-S-conjugates. Best-performing variant will be sequenced and the mutant enzymes will be expressed and purified.
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