Fig. 6: Application of the developed multilocus integration method in the biosynthesis of heme.
From: Efficient and markerless gene integration with SlugCas9-HF in Kluyveromyces marxianus
a The reconstituted biosynthetic pathway of heme and its precursors. The key precursor of heme (5-ALA) can be accumulated by the C4 pathway (the encoding genes are shown in green) and the C5 pathway (the encoding genes are shown in blue). A total of 11 heterologous genes to be integrated were divided into four panels and ligated to construct four multiple-gene cassettes, which could be integrated into any of the four loci or none at one step. b Heme production of newly formed single colonies isolated from the 6 top production colonies in primary screening. Error bars represent the mean ± SD of biological triplicates. c Red pigment accumulation in the five most productive newly formed single colonies isolated from the mixed colony 16.
