Fig. 1: HA accumulated in the cytosol of C. glutamicum expressing SzHAS derived from S. equi subsp. zooepidemicus.
A Schematics of Class I-R HAS. Class I-R HAS derived from Streptococcus, is a transmembrane protein that elongates and secretes HA. The cytosolic domain binds to activated nucleotide sugar monomers and facilitates glycosyltransferase reactions by alternately adding GlcA or GlcNAc to the growing HA chain. The HA chain elongates and translocates through the HAS pore, crossing the cell membrane until released. B PH, TEM, and SEM micrographs of C. glutamicum cells expressing SzHAS. PH microscopy observations were performed in triplicate and yielded similar results. C HA was detected in the cell pellets of C. glutamicum. Extracted ion chromatogram and mass spectra of HA analyzed by HPLC-MS. D Schematic showing HA accumulation in the cytosol and its effect on cell membrane integrity. E TEM micrographs of C. glutamicum cells retaining HA. F Growth and metabolism curves of C. glutamicum cells expressing SzHAS and SzHAS mutants SzHAS103A-105A (lacking HA synthesis ability) as the control. SzHAS-PI overexpressed SzHAS and PI synthase. G Comparison of membrane lipid composition between S. equi subsp. zooepidemicus WSH-24 and C. glutamicum ATCC13032. Data are expressed as mean ± S.D. from six (n = 6) independent biological replicates. H TEM micrographs of C. glutamicum cells expressing SzHAS and PI synthase. Cells cultivated for 32 h (late stationary phase). I Schematic diagram of the fatty acid synthesis of C. glutamicum. Statistical evaluation (p-value) was performed by a two-sided t test. The image shown is representative of n = 3 independent replicates of experiments with similar results. Source data are provided as a Source Data file.
