TABLE 1.

Sugar analysis of cell walls prepared from Cgl WT and the cg0847 conditional knockdown mutant

Cell walls were prepared and hydrolyzed with 2 m TFA as described (44), and alditol acetates derived thereof were subjected to GC/MS. The amounts of each monosaccharide in the samples are expressed relative to Rha set to be constant. Note that the 2 m TFA acid hydrolysis does not fully release the GlcNAc and Mur from the cell wall. An independent LC/MS-based method to measure cAG to PG ratio via the ratio MurNAc to MurNAc-6P (see Fig. 7) was also performed. The results are representative of two experiments performed on two independent culture batches.

Cgl strain	Rha	Ara	Man	Gal	Glc	GalNAc	Mur	GlcNAc
WT (1 mm IPTG)	3.1	78.4	1.6	23.9	3.7	22.6	15.2	41.2
cKD-0847 (1 mm IPTG)	3.1	99.5	4.1	34.6	10.4	9.3	13.0	38.1
WT (0 mm IPTG)	3.1	85.7	3.2	29.7	5.6	27.0	24.2	50.5
cKD-0847 (0 mm IPTG)	3.1	121.3	2.9	26.4	2.7	20.8	103.9	175.2