In this study, we purified a partially acetylated heteropolysaccharide (Ts1-1A) from the fruit bodies of Trametes sanguinea Lloyd through cold water extraction and serial chromatographic separation. The purified polysaccharide Ts1-1A (12.8 kDa) was characterized as a branched mannogalactofucan with a backbone of alternately connected 1,3-linked α-Fucp and 1,6-linked α-Galp, which was partially substituted by non-reducing end units of β-Manp at O-2 and O-3 positions of 1,6-linked α-Galp. Ts1-1A showed pronounced anti-human cytomegalovirus activity at the concentration of 200 and 500 μg/mL in systematical assessments including morphological changes, western blotting, qPCR, indirect immunofluorescence and tissue culture infective dose assays. Moreover, Ts1-1A exerted its antiviral activity at two distinct stages of viral proliferation manifesting as significantly inhibiting viral protein (IE1/2 and p52) expression and reducing viral gene (UL123, UL44 and UL32) replication in the HCMV-infected WI-38 cells. At viral attachment stage, Ts1-1A interacted with HCMV and prevented HCMV from attaching to its host cells. While at early phase of viral replication stage, Ts1-1A suppressed HCMV replication by downregulating NQO1 and HO-1 proteins related to oxidative stress as an antioxidant. To sum up, Ts1-1A is a promising anti-HCMV agent which could be developed for HCMV infection prevention and therapy

heteropolysaccharide, Structural characterization, oxidative stress, Trametes sanguinea, human cytomegalovirus, viral attachment

13C NMR data:
Linkage	Residue	C1	C2	C3	C4	C5	C6
6,3,6	aLFucp	98.4	68.5	77.6	73.4	68.9	15.8
6,3	aDGalp	98.0	69.3	68.9	70.2	73.5	66.6
6	aLFucp	98.4	68.5	77.6	73.4	68.9	15.8
2	bDManp	102.4	70.5	70.1	68.5	73.4	61.1
3	bDManp	102.4	70.5	70.1	68.5	73.4	61.1
	aDGalp	101.5	77.7	77.5	73.6	66.2	66.5


1H NMR data:
Linkage	Residue	H1	H2	H3	H4	H5	H6
6,3,6	aLFucp	4.95	3.73	3.88	3.57	4.10	1.13
6,3	aDGalp	4.90	4.06	4.14	3.78	3.67	3.54-3.55
6	aLFucp	4.95	3.73	3.88	3.57	4.10	1.13
2	bDManp	5.01	3.96	3.79	3.68	3.55	3.65-3.78
3	bDManp	5.01	3.96	3.79	3.68	3.55	3.65-3.78
	aDGalp	5.04	3.80	3.96	3.65	3.90	3.52-3.53


1H/13C HSQC data:
Linkage	Residue	C1/H1	C2/H2	C3/H3	C4/H4	C5/H5	C6/H6
6,3,6	aLFucp	98.4/4.95	68.5/3.73	77.6/3.88	73.4/3.57	68.9/4.10	15.8/1.13
6,3	aDGalp	98.0/4.90	69.3/4.06	68.9/4.14	70.2/3.78	73.5/3.67	66.6/3.54-3.55
6	aLFucp	98.4/4.95	68.5/3.73	77.6/3.88	73.4/3.57	68.9/4.10	15.8/1.13
2	bDManp	102.4/5.01	70.5/3.96	70.1/3.79	68.5/3.68	73.4/3.55	61.1/3.65-3.78
3	bDManp	102.4/5.01	70.5/3.96	70.1/3.79	68.5/3.68	73.4/3.55	61.1/3.65-3.78
	aDGalp	101.5/5.04	77.7/3.80	77.5/3.96	73.6/3.65	66.2/3.90	66.5/3.52-3.53

There is only one chemically distinct structure:

SVGMWSMILES3D molIonize

 

Warning = sub-repeat ending at aDGalp: large unit count (7) was reduced to 5
*O[C@@H]1[C@H](O)[C@H](C)O[C@@H](OC[C@H]2O[C@H](O[C@H]3[C@H](O)[C@H](OC[C@H]4O[C@H](O[C@H]5[C@H](O)[C@H](OC[C@H]6O[C@H](O[C@H]7[C@H](O)[C@H](OC[C@H]8O[C@H](O[C@H]9[C@H](O)[C@H](OC[C@H]%10O[C@H](O[C@H]%11[C@H](O)[C@H](OC[C@H]%12O[C@H](*)[C@H](O[C@@H]%13O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%13O)[C@@H](O[C@@H]%13O[C@H](CO)[C@@H](O)[C@H](O)[C@@H]%13O)[C@H]%12O)O[C@@H](C)[C@H]%11O)[C@H](O)[C@@H](O)[C@H]%10O)O[C@@H](C)[C@H]9O)[C@H](O)[C@@H](O)[C@H]8O)O[C@@H](C)[C@H]7O)[C@H](O)[C@@H](O)[C@H]6O)O[C@@H](C)[C@H]5O)[C@H](O)[C@@H](O)[C@H]4O)O[C@@H](C)[C@H]3O)[C@H](O)[C@@H](O)[C@H]2O)[C@H]1O

2173.98 g/mol (C₈₄H₁₄₀R₂O₆₄, R = next and previous repeats, not counted in mol weight)