AI Article Synopsis
- * A high-efficiency mutant (N212m) of the GjCCD4a gene, which catalyzes crocin production, was created, significantly boosting productivity by 25.08 times compared to the original gene.
- * The modified E. coli strains E57 and E579 successfully synthesized multiple crocins, yielding up to 11.50 mg/L and 8.43 mg/L, respectively, demonstrating a viable method for large-scale production using a glucose-based system.
Article Abstract
Crocins are bioactive natural products that rarely exist in plants. High costs and resource shortage severely limit its development and application. Synthetic biology studies on crocins are of considerable global interest. However, the lack of high-efficiency genetic tools and complex cascade biocatalytic systems have substantially hindered progress in crocin biosynthesis-related research. Based on mutagenesis, a high-efficiency GjCCD4a mutant (N212m) was constructed with a catalytic efficiency that was 25.08-fold higher than that of the wild-type. Solubilized GjCCD4a was expressed via fusion with an MBP tag. Moreover, N212m and ten other genes were introduced into Escherichia coli for the de novo biosynthesis of five crocins. The engineered E57 strain produced crocins III and V with a total yield of 11.50 mg/L, and the E579 strain produced crocins I-V with a total output of 8.43 mg/L at shake-flask level. This study identified a marvelous genetic element (N212m) for crocin biosynthesis and achieved its de novo biosynthesis in E. coli using glucose. This study provides a reference for the large-scale production of five crocins using E. coli cell factories.
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| http://dx.doi.org/10.1016/j.ijbiomac.2024.133985 | DOI Listing |
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