木质素结构单元对纤维素酶吸附的影响

doi:10.11707/j.1001-7488.LYKX20210766

Abstract

Abstract:

Objective: Using lignin with different structural units isolated from straw, reed and bamboo as the research material, the dynamic behavior between isolated lignin and enzyme adsorption was studied based on quartz crystal microbalance with dissipation (QCM-D) in situ and in real-time. The results obtained could provide targeted regulation for the efficient utilization of lignocellulose. Method: The chemical composition of raw materials was determined according to the standard method, the structural units of isolated lignin were characterized by infrared, alkaline nitrobenzene oxidation and gel permeability chromatography. The surface morphology of lignin film was detected by using an atomic force microscope, and the adsorption behavior of cellulase on lignin film was studied by QCM-D. Result: The isolated lignin was GSH type with high purity, the sugar content was about 10%, and their molecular weight was close, about 5 000. The nitrobenzene oxidation results indicated that structural units of isolated lignins were different, uncondensed units yield of bamboo lignin was highest, 444.4 g·kg^?1, and its S/G mole ratio was 1∶0.6. The lignin S/G ratio in reed was 1∶1.1, as well as rice straw. Bamboo lignin film showed the fastest adsorption rate of cellulase, and its adsorption amount was also the maximum via QCM-D analysis. The adsorption rate and maximum adsorption amount of cellulase in reed and straw lignin films were similar. After stopped the cellulase injection, the lignin film was rinsed with buffer solution while the cellulase attached to lignin was difficult to be eluted. All three lignin surfaces during cellulase adsorption had good viscoelasticity with little difference. Conclusion: Lignin present an obvious adsorption effect on cellulase, lignin with the same structural units, even from different sources, showed the same adsorption capacity of lignin to cellulase. Different structural units of lignin have different adsorption capacities for cellulase. Cellulase was hard to be removed because of its binding closely on the surface of lignin film. The lignin with higher S/G ratio showed more strong adsorption capacity to cellulase, which produced more ineffective adsorption in cellulase hydrolysis, and led to reduction of glucan conversion efficiency in enzymatic hydrolysis.

Key words: lignin, structural unit, enzyme adsorption, cellulose enzyme hydrolysis, QCM-D (quartz crystal microbalance with dissipation)

CLC Number:

S781.4

Chunyang Zou,Wenjuan Wu. Effect of Lignin Structural Unit on Cellulase Adsorption[J]. Scientia Silvae Sinicae, 2023, 59(6): 141-148.

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木质素 Lignin	葡聚糖 Glucan/ (g·kg^?1)	木聚糖 Xylan/ (g·kg^?1)	其他 Others/ (g·kg^?1)	总糖 Sum/ (g·kg^?1)
芦草 Phragmites australis	25.1±0.1	41.6±0.0	29.3±0.2	96.0±0.3
稻草 Oryza sativa	27.8±0.1	48.6±0.1	26.7±0.1	103.1±0.3
竹子 Bambusoideae	36.8±0.0	66.3±0.1	7.0±0.1	110.1±0.2

木质素 Lignin	数均分子质量 Number-average molecular weight (M_n)	重均分子质量 Mass average molar mass (M_w)	PDI (M_w / M_n)
芦草 Phragmites australis	5 456	10 418	1.91
稻草 Oryza sativa	6 118	12 265	2.00
竹子 Bambusoideae	5 043	10 831	2.15

木质素 Lignin	结构单元比例Molar ratio of structural units			结构单元得率 Yield of structural unit / (g·kg^?1)			总得率 Total yield/ (g·kg^?1)
木质素 Lignin	S	G	H	S	G	H	总得率 Total yield/ (g·kg^?1)
芦草 Phragmites australis	1	1.1	0.7	180.9±1.1	158.2±1.2	81.1±1.1	420.2±1.1
稻草 Oryza sativa	1	1.1	0.6	138.8±1.2	124.3±1.2	53.1±1.3	316.2±1.1
竹子 Bambusoideae	1	0.6	0.4	248.2±1.2	129.9±1.3	67.3±1.3	444.4±1.2

木质素 Lignin	葡聚糖 Glucan(%)	木聚糖 Xylan (%)	总糖 Sum (%)
芦草 Phragmites australis	86.6	62.1	75.1
稻草 Oryza sativa	81.0	69.3	66.1
竹子 Bambusoideae	71.5	66.2	68.9

木质素 Lignin	粗糙度 Roughness/nm
芦草 Phragmites australis	1.67±0.09
稻草 Oryza sativa	1.76±0.07
竹子 Bambusoideae	2.16±0.06

Effect of Lignin Structural Unit on Cellulase Adsorption

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木质素薄膜 Lignin films	$- {M}_{\mathrm{m}\mathrm{a}\mathrm{x}} $ /Hz	1/ $ \tau $ /min^?1	R²	最大吸附量 Maximum adsorption capacity / (ng·cm^?2)
芦草 Phragmites australis	48.1	0.06	0.93	283.9
稻草 Oryza sativa	48.1	0.06	0.95	283.6
竹子 Bambusoideae	50.7	0.07	0.97	298.8

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