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Mutagenesis Services> | … For in vitro assays, the CsCCD2 gene was codon-optimized and synthesized by GenScript (Nanjing, China) (Table S5) and then cloned into the NcoI/XhoI site of pET28a for expression in E. coli. And the codons used for site-directed mutagenesis are also listed in Table S3. … | Get A Quote |
Crocetin, a high-value apocarotenoid in saffron, is widely applied to the fields of food and medicine. However, the existing method of obtaining crocetin through large-scale cultivation is far from meeting the market demand. Microbial synthesis of crocetin is a potential alternative to traditional resources, and it is found that carotenoid cleavage dioxygenase (CCD) is the critical enzyme to synthesize crocetin. So, in this study, we used "hybrid-tunnel" engineering to obtain variants of -derived CCD2, essential for zeaxanthin conversion into crocetin, with a broader substrate specificity and higher catalytic efficiency. Variants including S323A, with a lower charge bias and a larger tunnel size than the wild-t... More