Fig. 7

The production of lycopene by engineered O. polymorpha and detection of lycopene. A The lycopene biosynthesis pathway in O. polymorpha. B A schematic representation of the integration of lycopene biosynthesis-related genes CrtE, CrtYB, and into three different neutral sites NS18, NS12, and 3NS2 in the genome using the CRISPR-Cpf1 system in one-step. C Verification of transformants using colony PCR. Five clones were selected and colony PCR was employed to confirm the integration of the three genes at their designated locations. The sizes of PCR products were 3465 bp for CrtE, 4356 bp for CrtYB, and 4083 bp for CrtI, indicating precise integration into the specified genomic sites (M: marker; lanes 1, 7, 13 were negative control; lanes 2–6 were products pf CrtYB; lanes 8–12 were products of CrtI; lanes 14–18 were products of CrtE.). D Analysis of lycopene extracted from F41 strain after 3 days of fermentation in BMMY media was performed using HPLC