Comparison of four glycosyl residue composition methods for effectiveness in detecting sugars from cell walls of dicot and grass tissues

Biswal, Ajaya K.; Tan, Li; Atmodjo, Melani A.; DeMartini, Jaclyn; Gelineo-Albersheim, Ivana; Hunt, Kimberly; Black, Ian M.; Mohanty, Sushree S.; Ryno, David; Wyman, Charles E.; Mohnen, Debra

doi:10.1186/s13068-017-0866-1

Biotechnology for Biofuels and Bioproducts

Table 2 Comparison of different glycosyl residue analysis methods for μg monosaccharide quantified per mg of AIR from three different biofuel feedstock biomass samples from three different species

From: Comparison of four glycosyl residue composition methods for effectiveness in detecting sugars from cell walls of dicot and grass tissues

Biomass-method	μg glycosyl residue/mg biomass AIR
Biomass-method	Ara	Rha	Fuc	Xyl	GlcA	GalA	Man	Gal	Glc	Total neutral	Total acidic	Total
Populus wood
Alditol acetate	6.7 ± 0.6	3.0 ± 0.3	0.4 ± 0.01	175.3 ± 12.3	-	-	12.1 ± 0.9	9.4 ± 0.8	20.6 ± 1.5	227.5		227.4
Uronic acid*	-	-	-	-	37.5 ± 1.3		-	-	-		37.5
Alditol acetate + uronic acid**	6.7 ± 0.6^a	3.0 ± 0.3^a	0.4 ± 0.01	175.3 ± 12.3^a	37.5 ± 1.3		12.1 ± 0.9^a	9.4 ± 0.8^a	20.6 ± 1.5	227.5^a	37.5^b	264.9^a
Carbodiimide	9.0 ± 0.5^b	4.0 ± 0.5^a	0.5 ± 0.01	190.1 ± 10.2^c	1.8 ± 0.1	16.4 ± 0.9	13.0 ± 1.0^a	12.0 ± 0.8^b	22.5 ± 1.8	251.1^b	18.2^a	269.3^a
TMS	9.3 ± 0.6^b	4.2 ± 0.4^a	0.4 ± 0.01	197.6 ± 7.9^c	2.0 ± 0.2	18.1 ± 1.1	13.2 ± 0.9^a	10.7 ± 0.6^a	20.9 ± 1.3	256.3^b	20.1^a	276.4^b
HPAEC	11.8 ± 0.8^c	5.0 ± 0.6^b	0.5 ± 0.01	185.9 ± 8.5^b	2.9 ± 0.2	17.9 ± 1.3	14.6 ± 0.8^b	12.3 ± 0.9^b	21.3 ± 1.9	251.4^b	20.8^a	272.2^b
Rice stem
Alditol acetate	38.9 ± 3.1	3.0 ± 0.3	1.0 ± 0.02	223.6 ± 9.9	-	-	2.0 ± 0.2	15.1 ± 1.3	29.3 ± 3.1	312.9		312.9
Uronic acid*	-	-	-	-	11.9 ± 0.8		-	-	-		11.9
Alditol acetate + uronic acid**	38.9 ± 3.1^b	3.0 ± 0.3^b	1.0 ± 0.02^b	223.6 ± 9.9^a	11.9 ± 0.8		2.0 ± 0.2	15.1 ± 1.3	29.3 ± 3.1	312.9^a	11.9^b	324.8^a
Carbodiimide	41.3 ± 3.8^b	2.3 ± 0.3^b	0.4 ± 0.01^a	225.6 ± 16.2^b	0.9 ± 0.01	8.0 ± 0.6^b	2.0 ± 0.2	16.8 ± 0.9	31.3 ± 2.9	319.7^b	8.9^a	328.6^a
TMS	39.5 ± 2.7^b	2.0 ± 0.2^a	0.8 ± 0.01^b	230.3 ± 10.3^b	1.0 ± 0.01	8.9 ± 0.5^b	1.5 ± 0.1	15.8 ± 1.4	30.8 ± 2.6	320.7^b	9.9^a	330.6^a
HPAEC	35.9 ± 3.4^a	2.5 ± 0.2^b	0.8 ± 0.01^b	242.7 ± 14.7^c	1.9 ± 0.02	6.5 ± 0.5^a	2.9 ± 0.3	14.6 ± 0.7	32.8 ± 2.2	332.2^c	8.4^a	340.5^b
Switchgrass tiller
Alditol acetate	52.0 ± 2.1	3.3 ± 0.4	0.7 ± 0.01	265.2 ± 9.6	-	-	1.8 ± 0.02	16.5 ± 2.0	22.5 ± 2.1	362.0		361.9
Uronic acid*	-	-	-	-	4.7 ± 0.4		-	-	-		4.7
Alditol acetate + uronic acid**	52.0 ± 2.1	3.3 ± 0.4^c	0.7 ± 0.01	265.2 ± 9.6^a	4.7 ± 0.4		1.8 ± 0.02^b	16.5 ± 2.0^b	22.5 ± 2.1	362.0^a	4.7	366.6^a
Carbodiimide	50.3 ± 2.6	2.3 ± 0.2^b	0.4 ± 0.01	270.4 ± 12.7^b	0.7 ± 0.01	3.0 ± 0.4^a	1.4 ± 0.02^a	14.3 ± 0.8^b	22.4 ± ± 1.9	361.5^a	3.7	365.1^a
TMS	52.7 ± 3.2	2.6 ± 0.2^b	0.3 ± 0.01	278.8 ± ± 17.3^c	0.8 ± 0.01	3.4 ± 0.4^b	1.0 ± ± 0.01^a	13.1 ± 1.1^a	22.0 ± 2.3	370.5^b	4.2	374.7^b
HPAEC	53.6 ± 3.7	1.5 ± 0.1^a	0.5 ± 0.01	270.6 ± 14.3^b	0.9 ± 0.01	2.9 ± 0.2^a	2.1 ± 0.01^c	15.9 ± 0.9^b	20.9 ± 2.6	365.1^a	3.8	368.9^a

Data are means of three biological replicates ± standard deviation, with two technical replicates for alditol acetate, carbodiimide, and TMS methods (n = 6), three technical replicates for uronic acid assay (n = 9), and a single technical replicate for HPAEC method (n = 3). Different letters in superscript following the values indicate significant differences between the amounts (“a” represents the lowest amount) of a particular (or total) sugar residue in the cell walls of a species across different methods (and not between different types of sugars within that biomass sample analyzed using a particular method). Statistics are one-way analysis of variance (ANOVA) followed by Tukey’s multiple comparison tests with significant P value <0.05
* Uronic acid assay data shown here are from analysis using TFA hydrolysis
** Alditol acetate + uronic acid data shown here are the combined data of the respective individual assays

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