辣根过氧化物酶改性木质素磺酸钠的结构特征及吸附分散性能

doi:10.7503/cjcu20130766

摘要/Abstract

摘要：

在室温及水溶液体系中, 采用辣根过氧化物酶(HRP)对亚硫酸法制浆造纸废液的副产物木质素磺酸钠(木钠)进行改性, 通过凝胶渗透色谱、电位滴定、红外光谱和核磁共振波谱等表征了HRP改性木钠的结构. 结果表明, HRP可以有效聚合木钠大分子, 调节HRP的用量, 得到不同分子量的木钠产品, 当HRP浓度为6 g/L时, 可使木钠分子量增大155%. HRP可氧化木钠分子上的酚羟基变成苯氧自由基, 该自由基可直接交联, 也可转移到酚羟基的邻位或对位再发生聚合作用, 其聚合方式主要为β-O-4'及β-β'连接. HRP改性还可使木钠磺化度增加27%. 采用静电逐层自组装技术研究了HRP改性对木钠吸附特征的影响, 结果表明, 经HRP改性后, 木钠在平板上的吸附量增大; 对TiO₂浆体的分散稳定性能也得到改善, 这主要是因为分子量增大, 空间位阻作用增强; 磺化度增大, 静电排斥作用增强, 从而使TiO₂颗粒更好地分散在水中.

关键词: 辣根过氧化物酶, 木质素磺酸钠, 分子量分布, 结构特征, 吸附分散性能

Abstract:

A novel and efficient polymerization of lignosulofonate(by-products from sulfite pulping process) was investigated in the present study. The ability of commercial horseradish peroxidase(HRP) for polymerizing sodium lignosulfonate(SL) was investigated in aqueous solution at room temperature. The gel permeation chromatography(GPC), potentiometric titration, infrared spectroscopy(IR), and ¹H nuclear magnetic resonance(¹H NMR) were used the structural characterization of SL. GPC results showed a significant increase in molecular weight(M_w) of SL. With different dosages of HRP added, SLs with different M_w could be obtained. When 6 g/L HRP was added, the M_w of SL could increase by 155%. During the HRP incubation, phenolic groups were oxidized to phenoxyl radicals. These radicals could undergo radical-couplings directly. On the other hand, unbound electron could transfer to the para- and ortho-carbon of the phenols and β-carbon of the branch, undergoing radical-couplings. Among the radical-couplings, β-O-4' and β-β' were the predominant. Moreover, the sulfonic group increased by 27% after HRP incubation. The adsorption of SL by HRP incubation was studied by electrostatic self-assembly technology. After HRP incubation, the adsorption amount of SL increased significantly. Further, because of the increase of M_w and sulfonic group content, resulting in the stronger steric hindrance and electronic repulsion, the dispersion ability of SL on TiO₂ slurry was improved.

Key words: Horseradish peroxidase, Sodium lignosulfonate, Molecular weight distribution, Structural characterization, Adsorption and dispersion property

中图分类号:

TrendMD:

周海峰, 杨东杰, 邱学青, 伍晓蕾. 辣根过氧化物酶改性木质素磺酸钠的结构特征及吸附分散性能. 高等学校化学学报, 2014, 35(4): 895.

ZHOU Haifeng, YANG Dongjie, QIU Xueqing, WU Xiaolei. Structural Characterization, Adsorption and Dispersion Properties of Sodium Lignosulfonate by Horseradish Peroxidase Incubation^†. Chem. J. Chinese Universities, 2014, 35(4): 895.

图/表 10

Table 1 Reaction conditions and molecular weight distribution of SL by HRP incubation

Sample	Concentration of HRP/(g·L^-1)	Incubation time/h	10^-3 M_n	10^-4 M_w	M_w/M_n
D748	0	0	4.8	1.40	2.92
SL	0	0	2.7	0.99	3.69
HRP-SL 1	0.2	12	2.9	1.13	3.96
HRP-SL 2	0.5	12	3.3	1.42	4.25
HRP-SL 3	1	12	3.5	1.63	4.62
HRP-SL 4	3	12	3.8	1.90	5.03
HRP-SL 5	6	12	4.6	2.50	5.47
HRP-SL 6	1	0.33	3.5	1.61	4.58
HRP-SL 7	1	0.67	3.5	1.63	4.60
HRP-SL 8	1	1.0	3.6	1.65	4.63
HRP-SL 9	1	36	3.6	1.66	4.63
HRP-SL 10	6	0.33	4.6	2.48	5.45
HRP-SL 11	6	0.67	4.5	2.49	5.48
HRP-SL 12	6	1.0	4.6	2.51	5.47
HRP-SL 13	6	36	4.6	2.52	5.49

Fig.1 Effect of concentration of HRP on molecular weight distribution of SL by HRP incubation (A) Molecular weight distribution; (B) relationship of molecular weight and concentration of HRP.

Table 2 Phenolic and sulfonic group content of SL by HRP incubation

Sample	Concentration of phenolic group/(mmol·g^-1)	Concentration of sulfonic group/(mmol·g^-1)
SL	2.35	1.30
HRP-SL 6	1.72	1.42
HRP-SL 7	1.70	1.43
HRP-SL 8	1.74	1.42
HRP-SL 3	1.70	1.44
HRP-SL 9	1.71	1.41
HRP-SL 10	1.33	1.64
HRP-SL 11	1.32	1.63
HRP-SL 12	1.32	1.61
HRP-SL 5	1.34	1.64
HRP-SL 13	1.32	1.65

Fig.2 IR spectra of SL by HRP incubation a. SL; b. HRP-SL 3; c. HRP-SL 5.

Table 3 IR bands assignment of SL and the ratio of characteristic peak at 1512 cm-1

Peak intensity ratio	Assignment	SL	HRP-SL 3	HRP-SL 5
A₃₄₃₆/A₁₅₁₂	OH stretching in phenolic and aliphatic structures	1.588	1.430	1.347
A₂₉₃₇/A₁₅₁₂	C—H vibration in —CH₃ and —CH₂	1.002	0.987	0.969
A₂₈₅₀/A₁₅₁₂	C—H vibration in CH₃O—	0.962	0.947	0.770
A₁₇₀₃/A₁₅₁₂	C=O vibration in unconjugated ketone, carbonyl and ester groups	0.812	0.846	0.861
A₁₆₀₄/A₁₅₁₂	Aromatic skeleton vibrations	1.242	1.181	1.174
A₁₄₆₂/A₁₅₁₂	C—H deformations band of asymmetric methyl and methylene	0.948	0.921	0.872
A₁₄₂₁/A₁₅₁₂	Aromatic skeleton vibrations combined with C—H in plane deformations	0.995	0.996	0.989
A₁₃₆₆/A₁₅₁₂	Aliphatic C—H stretching in methyl groups and phenolic hydroxyl	0.961	0.906	0.842
A₁₁₆₄/A₁₅₁₂	G ring plus C—O stretching	1.686	1.437	1.318
A₁₀₇₆/A₁₅₁₂	S ring and C—O stretching	1.409	1.302	1.264
A₁₀₄₄/A₁₅₁₂	S=O stretching	1.576	1.703	1.824

Fig.3 1H NMR spectra of SL by HRP incubation a. SL; b. HRP-SL 5.

Table 4 Signal assignment in 1H NMR spectra of SL and results for quantification of functional groups(DMSO signal intensity as reference)

δ	Assignment	Amount(DMSO^-1)
δ	Assignment	SL	HRP-SL 5
7.75—7.20	Aromatic protons in positions C-2 and C-6 with C=O group	1.05	1.16
7.20—6.80	H₂, H₅, H₆ in guaiacyl units(G)	1.00	0.83
6.80—6.20	H₂, H₆ in syringyl units(S)	0.56	0.51
5.75—5.25	H_α, H_β in β-β' structures	0.89	1.06
5.00—4.75	H_α in β-O-4' structures	0.73	0.95
4.75—4.07	H_β, H_γ in β-O-4' structures	2.74	4.69
4.00—3.50	H in methoxyls	5.66	3.57
3.20—2.90	$H β$ in β-β' structures	0.61	0.76
2.56—2.44	DMSO	1.00	1.00
2.40—2.10	H in aromatic acetates	0.50	0.58
1.95—1.85	H in aliphatic acetates	0.19	0.27
1.60—0.70	Aliphatic H	1.29	2.27

Table 4 Signal assignment in 1H NMR spectra of SL and results for quantification of functional groups(DMSO signal intensity as reference)

δ	Assignment	Amount(DMSO^-1)
δ	Assignment	SL	HRP-SL 5
7.75—7.20	Aromatic protons in positions C-2 and C-6 with C=O group	1.05	1.16
7.20—6.80	H₂, H₅, H₆ in guaiacyl units(G)	1.00	0.83
6.80—6.20	H₂, H₆ in syringyl units(S)	0.56	0.51
5.75—5.25	H_α, H_β in β-β' structures	0.89	1.06
5.00—4.75	H_α in β-O-4' structures	0.73	0.95
4.75—4.07	H_β, H_γ in β-O-4' structures	2.74	4.69
4.00—3.50	H in methoxyls	5.66	3.57
3.20—2.90	$H β$ in β-β' structures	0.61	0.76
2.56—2.44	DMSO	1.00	1.00
2.40—2.10	H in aromatic acetates	0.50	0.58
1.95—1.85	H in aliphatic acetates	0.19	0.27
1.60—0.70	Aliphatic H	1.29	2.27

Fig.4 UV-Vis spectra of SL/PDAC multilayers of SL-Control

Fig.5 Absorbance of SL/PDAC multilayers at 280 nm varying with bilayer numbers by HRP incubation

Fig.6 Effect of SL by HRP incubation on particle size of TiO2 slurry a. No dispersant; b. SL; c. HRP-SL 3; d. HRP-SL 5.

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