Microbial Resources and Application Laboratory, School of Food Science and Engineering, Hefei University of Technology, Hefei, China
Exopolysaccharide of Lachnum YM130 (LEP) was purified by diethylaminoethyl cellulose 52 and Sepharose CL-6B column chromatography. LEP-2a was identified to be a homogeneous component with an average molecular weight of 1.31 x 10(6) Da, which was consisted of mannose and galactose in a molar ratio of 3.8:1.0. The structure of LEP-2a was characterized by methylation analysis, FT-IR analysis, and NMR analysis. Results indicated that LEP-2a was a galactomannan with a backbone, composed of 1,2-linked-alpha-D-Manp, 1,2,6-linked-alpha-D-Manp, 1,3,4-linked-alpha-D-Manp, and 1,3-linked-beta-D-Galp, which was substituted at O-2, O-3, O-4, and O-6 by branches. In vitro antitumor activity assay proved that LEP-2a could significantly enhance the inhibitory effectiveness of 5-FU on Hela cells at the concentrations of 100, 200, 300, and 400 mu g/mL. The above results suggested that LEP-2a could be seen as a potential source for developing novel antineoplastic agents.
fig. 5, LEP-2a, table 2
13C NMR, 1H NMR, GC-MS, HPLC, statistical analysis, DEAE, methylation analysis, dialysis, cell viability assay, HPGPC, precipitation, phenol-sulfuric acid assay, HMBC, antitumor activity assay, DEPT, centrifugation, COSY, HSQC, filtration, FT-IR, TFA hydrolysis, concentration
decreases the viability of SGC7901, SMMC-7721, Hela cells; enhances the inhibitory effect of 5-FU on Hela cells
13C NMR data:
Linkage Residue C1 C2 C3 C4 C5 C6
6,3,2,3,4,2,4 aDManp 100.53 78.37 70.35 66.50 73.18 60.92
6,3,2,3,4,2,3 aDManp 102.25 69.98 70.18 66.58 73.25 60.98
6,3,2,3,4,2 aDManp 102.13 69.55 78.37 70.53 73.05 60.98
6,3,2,3,4,6,3,3 aDManp 102.25 69.98 70.18 66.58 73.25 60.98
6,3,2,3,4,6,3,4 bDGalp 100.26 70.16 73.17 70.49 72.86 60.95
6,3,2,3,4,6,3 aDManp 102.13 69.55 78.37 70.53 73.05 60.98
6,3,2,3,4,6 bDGalp 99.38 69.97 78.59 70.463 76.17 60.94
6,3,2,3,4 aDManp 98.26 78.69 70.43 66.87 73.24 65.55
6,3,2,3,3 aDManp 102.25 69.98 70.18 66.58 73.25 60.98
6,3,2,3 aDManp 102.13 69.55 78.37 70.53 73.05 60.98
6,3,2,4,3,3 aDManp 102.25 69.98 70.18 66.58 73.25 60.98
6,3,2,4,3,4,3 aDManp 102.25 69.98 70.18 66.58 73.25 60.98
6,3,2,4,3,4,4 bDGalp 100.26 70.16 73.17 70.49 72.86 60.95
6,3,2,4,3,4 aDManp 102.13 69.55 78.37 70.53 73.05 60.98
6,3,2,4,3 aDManp 102.13 69.55 78.37 70.53 73.05 60.98
6,3,2,4 bDGalp 99.38 69.97 78.59 70.43 73.17 60.94
6,3,2 aDManp 102.13 69.55 78.37 70.53 73.05 60.98
6,3 aDManp 100.53 78.37 70.35 66.50 73.18 60.92
6 bDGalp 99.38 69.97 78.59 70.43 73.17 60.94
2,2,3 aDManp 102.25 69.98 70.18 66.58 73.25 60.98
2,2,4 bDGalp 100.26 70.16 73.17 70.49 72.86 60.95
2,2 aDManp 102.13 69.55 78.37 70.53 73.05 60.98
2 aDManp 100.53 78.37 70.35 66.50 73.18 60.92
aDManp 98.26 78.69 70.43 66.87 73.24 65.55
1H NMR data:
Linkage Residue H1 H2 H3 H4 H5 H6
6,3,2,3,4,2,4 aDManp 5.27 4.10 3.87 3.72 3.72 3.73-3.85
6,3,2,3,4,2,3 aDManp 5.13 4.06 3.89 3.74 3.75 3.74-3.87
6,3,2,3,4,2 aDManp 5.03 4.21 3.93 3.81 3.61 3.73-3.86
6,3,2,3,4,6,3,3 aDManp 5.13 4.06 3.89 3.74 3.75 3.74-3.87
6,3,2,3,4,6,3,4 bDGalp 4.86 3.89 3.74 3.72 3.58 3.75-3.84
6,3,2,3,4,6,3 aDManp 5.03 4.21 3.93 3.81 3.61 3.73-3.86
6,3,2,3,4,6 bDGalp 4.89 3.98 3.88 3.70 3.64 3.76-3.84
6,3,2,3,4 aDManp 5.10 4.01 3.85 3.66 3.73 3.96
6,3,2,3,3 aDManp 5.13 4.06 3.89 3.74 3.75 3.74-3.87
6,3,2,3 aDManp 5.03 4.21 3.93 3.81 3.61 3.73-3.86
6,3,2,4,3,3 aDManp 5.13 4.06 3.89 3.74 3.75 3.74-3.87
6,3,2,4,3,4,3 aDManp 5.13 4.06 3.89 3.74 3.75 3.74-3.87
6,3,2,4,3,4,4 bDGalp 4.86 3.89 3.74 3.72 3.58 3.75-3.84
6,3,2,4,3,4 aDManp 5.03 4.21 3.93 3.81 3.61 3.73-3.86
6,3,2,4,3 aDManp 5.03 4.21 3.93 3.81 3.61 3.73-3.86
6,3,2,4 bDGalp 4.89 3.98 3.88 3.70 3.64 3.76-3.84
6,3,2 aDManp 5.03 4.21 3.93 3.81 3.61 3.73-3.86
6,3 aDManp 5.27 4.10 3.87 3.72 3.72 3.73-3.85
6 bDGalp 4.89 3.98 3.88 3.70 3.64 3.76-3.84
2,2,3 aDManp 5.13 4.06 3.89 3.74 3.75 3.74-3.87
2,2,4 bDGalp 4.86 3.89 3.74 3.72 3.58 3.75-3.84
2,2 aDManp 5.03 4.21 3.93 3.81 3.61 3.73-3.86
2 aDManp 5.27 4.10 3.87 3.72 3.72 3.73-3.85
aDManp 5.10 4.01 3.85 3.66 3.73 3.96
1H/13C HSQC data:
Linkage Residue C1/H1 C2/H2 C3/H3 C4/H4 C5/H5 C6/H6
6,3,2,3,4,2,4 aDManp 100.53/5.27 78.37/4.10 70.35/3.87 66.50/3.72 73.18/3.72 60.92/3.73-3.85
6,3,2,3,4,2,3 aDManp 102.25/5.13 69.98/4.06 70.18/3.89 66.58/3.74 73.25/3.75 60.98/3.74-3.87
6,3,2,3,4,2 aDManp 102.13/5.03 69.55/4.21 78.37/3.93 70.53/3.81 73.05/3.61 60.98/3.73-3.86
6,3,2,3,4,6,3,3 aDManp 102.25/5.13 69.98/4.06 70.18/3.89 66.58/3.74 73.25/3.75 60.98/3.74-3.87
6,3,2,3,4,6,3,4 bDGalp 100.26/4.86 70.16/3.89 73.17/3.74 70.49/3.72 72.86/3.58 60.95/3.75-3.84
6,3,2,3,4,6,3 aDManp 102.13/5.03 69.55/4.21 78.37/3.93 70.53/3.81 73.05/3.61 60.98/3.73-3.86
6,3,2,3,4,6 bDGalp 99.38/4.89 69.97/3.98 78.59/3.88 70.463/3.70 76.17/3.64 60.94/3.76-3.84
6,3,2,3,4 aDManp 98.26/5.10 78.69/4.01 70.43/3.85 66.87/3.66 73.24/3.73 65.55/3.96
6,3,2,3,3 aDManp 102.25/5.13 69.98/4.06 70.18/3.89 66.58/3.74 73.25/3.75 60.98/3.74-3.87
6,3,2,3 aDManp 102.13/5.03 69.55/4.21 78.37/3.93 70.53/3.81 73.05/3.61 60.98/3.73-3.86
6,3,2,4,3,3 aDManp 102.25/5.13 69.98/4.06 70.18/3.89 66.58/3.74 73.25/3.75 60.98/3.74-3.87
6,3,2,4,3,4,3 aDManp 102.25/5.13 69.98/4.06 70.18/3.89 66.58/3.74 73.25/3.75 60.98/3.74-3.87
6,3,2,4,3,4,4 bDGalp 100.26/4.86 70.16/3.89 73.17/3.74 70.49/3.72 72.86/3.58 60.95/3.75-3.84
6,3,2,4,3,4 aDManp 102.13/5.03 69.55/4.21 78.37/3.93 70.53/3.81 73.05/3.61 60.98/3.73-3.86
6,3,2,4,3 aDManp 102.13/5.03 69.55/4.21 78.37/3.93 70.53/3.81 73.05/3.61 60.98/3.73-3.86
6,3,2,4 bDGalp 99.38/4.89 69.97/3.98 78.59/3.88 70.43/3.70 73.17/3.64 60.94/3.76-3.84
6,3,2 aDManp 102.13/5.03 69.55/4.21 78.37/3.93 70.53/3.81 73.05/3.61 60.98/3.73-3.86
6,3 aDManp 100.53/5.27 78.37/4.10 70.35/3.87 66.50/3.72 73.18/3.72 60.92/3.73-3.85
6 bDGalp 99.38/4.89 69.97/3.98 78.59/3.88 70.43/3.70 73.17/3.64 60.94/3.76-3.84
2,2,3 aDManp 102.25/5.13 69.98/4.06 70.18/3.89 66.58/3.74 73.25/3.75 60.98/3.74-3.87
2,2,4 bDGalp 100.26/4.86 70.16/3.89 73.17/3.74 70.49/3.72 72.86/3.58 60.95/3.75-3.84
2,2 aDManp 102.13/5.03 69.55/4.21 78.37/3.93 70.53/3.81 73.05/3.61 60.98/3.73-3.86
2 aDManp 100.53/5.27 78.37/4.10 70.35/3.87 66.50/3.72 73.18/3.72 60.92/3.73-3.85
aDManp 98.26/5.10 78.69/4.01 70.43/3.85 66.87/3.66 73.24/3.73 65.55/3.96
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