[1]王锐,辛东林,张军华*.木质素降解产物对纤维素酶和木聚糖酶水解的抑制[J].林业工程学报,2019,4(04):78-84.[doi:10.13360/j.issn.2096-1359.2019.04.012]
 WANG Rui,XIN Donglin,ZHANG Junhua*.Inhibitory effects of vanillin, 4-hydroxybenzaldehyde and syringaldehyde on cellulases and xylanases[J].Journal of Forestry Engineering,2019,4(04):78-84.[doi:10.13360/j.issn.2096-1359.2019.04.012]
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木质素降解产物对纤维素酶和木聚糖酶水解的抑制()
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《林业工程学报》[ISSN:1001-8081/CN:32-1160/S]

卷:
4
期数:
2019年04期
页码:
78-84
栏目:
生物质能源与材料
出版日期:
2019-07-09

文章信息/Info

Title:
Inhibitory effects of vanillin, 4-hydroxybenzaldehyde and syringaldehyde on cellulases and xylanases
文章编号:
2096-1359(2019)04-0078-07
作者:
王锐辛东林张军华*
西北农林科技大学林学院,陕西 杨凌 712100
Author(s):
WANG Rui XIN Donglin ZHANG Junhua*
College of Forestry, Northwest A & F University, Yangling 712100, Shaanxi, China
关键词:
木质素降解产物 抑制 酶水解 纤维素酶 木聚糖酶
Keywords:
lignin-derived phenolic compounds inhibition enzymatic hydrolysis cellulase xylanase
分类号:
S785; TQ353
DOI:
10.13360/j.issn.2096-1359.2019.04.012
文献标志码:
A
摘要:
在稀酸等化学预处理过程中,木质素会降解生成酚类等产物进入后续糖化阶段,研究表明这些木质素降解产物会抑制木质纤维降解酶的水解效率,然而其抑制机制尚不清楚。笔者选择了3种典型的木质素降解产物:香草醛、4-羟基苯甲醛和丁香醛,考察了它们对商品纤维素酶和木聚糖酶,以及单一关键纤维素酶组分和β-木糖苷酶水解的影响,并探讨其抑制规律。实验结果表明,这3种木质素降解产物对纤维素酶和木聚糖酶的水解均有抑制,其抑制能力随降解产物浓度的增加而增强。当3种木质素降解产物的质量浓度为10 mg/mL时,纤维素酶水解微晶纤维素48 h的葡萄糖得率由71.17%分别减少到33.80%、29.52%和32.03%,说明这3种木质素降解产物对纤维素酶的抑制作用差异不明显。3种木质素降解产物对β-葡萄糖苷酶水解纤维二糖的效率没有影响,但是会强烈抑制外切葡聚糖酶CBH I的酶活。当木质素降解产物的质量浓度为2 mg/mL时,与未添加木质素降解产物的酶活相比,CBH I酶活分别降低至79.64%、86.76%和71.89%,抑制强弱顺序为:丁香醛>香草醛>4-羟基苯甲醛。此外,3种木质素降解产物对木聚糖酶和β-木糖苷酶的抑制强弱顺序均为:4-羟基苯甲醛>香草醛>丁香醛,当3种木质素降解产物的质量浓度为10 mg/mL时,木聚糖酶水解木聚糖48 h的木糖得率由57.28%分别减少到12.26%、20.16%和30.43%。抑制动力学试验表明,4-羟基苯甲醛对CBH I的抑制属于竞争性抑制,对β-木糖苷酶的抑制属于非竞争性抑制。
Abstract:
Biorefinery of lignocellulosic materials to liquid fuels is regarded as a promising way to produce renewable energy, but the biorefinery process is still not commercially viable due to the recalcitrance of cellulosic materials.Biomass-disrupting pretreatment is recognized as the most efficient method to break the recalcitrance of lignocellulosic materials and make them easily accessible to enzymes.During the pretreatment process, especially in dilute acid or steam explosion pretreatments, part of lignin in lignocelluloses is converted into phenolic compounds, which affects the following enzymatic hydrolysis.These phenolic compounds are found to be inhibitors of cellulases, but the detailed mechanism behind the inhibition is still unclear.In this paper, the effects of three typical lignin-derived phenolic compounds(vanillin, 4-hydroxybenzaldehydeand syringaldehyde)on the hydrolytic action of commercial cellulases, xylanase, and individual cellulase and β-xylosidase were investigated.The results indicated that all the three phenolic compounds were concentration-dependent inhibitors of cellulases and xylanase and could strongly inhibit the cellulases or xylanase at a high concentration(10 mg/mL).During the hydrolysis of Avicel by cellulases, the addition of 10 mg/mL vanillin, 4-hydroxybenzaldehyde and syringaldehydeclearly decreased glucose yields from 71.17% to 33.80%, 29.52% and 32.03%, respectively, indicating that the inhibitory effects of the three phenolic compounds on cellulases did not have obvious differences.The inhibitory effect was partly attributed to the negative effect on cellobiohydrolase I(CBH I), but not on β-glucosidase, even at the concentration of 10 mg/mL.The addition of 2 mg/mL vanillin, 4-hydroxybenzaldehyde and syringaldehyde could decrease the CBH I hydrolytic capacity from 100% to 79.64%, 86.76% and 71.89%, respectively, which indicated that syringaldehyde had the strongest inhibitory effect on CBH I among these three inhibitors, followed by vanillin and 4-hydroxybenzaldehyde.Kinetic experiments showed that 4-hydroxybenzaldehyde was a competitive inhibitor of the CBH I from Thermoascusaurantiacus.However, the order of their inhibitory effects on xylanase was the opposite.4-Hydroxybenzaldehyde was the strongest inhibitor of xylanase, and syringaldehyde exhibited the least inhibitory effect among the three phenolic compounds.The addition of 10 mg/mL vanillin, 4-hydroxybenzaldehyde and syringaldehyde decreased the xylose yields from 57.28% to 20.16%, 12.26%, and 30.43%, respectively, in the xylan hydrolysis by xylanase for 48 h.The three phenolic compounds could also strongly inhibit the β-xylosidase, and the order of the inhibitory effect on β-xylosidase was 4-hydroxybenzaldehyde>vanillin >syringaldehyde.Kinetic experiments showed that 4-hydroxybenzaldehyde was a non-competitive inhibitor of the β-xylosidase from Selenomonasruminantium.The results provided some valuable information for understanding the inhibition of lignin-derived phenolics on cellulases and xylanases.

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备注/Memo

备注/Memo:
收稿日期:2018-10-02 修回日期:2019-05-13
基金项目:国家自然科学基金(31670598)。
作者简介:王锐,男,硕士,研究方向为生物炼制与生物能源转化。通信作者:张军华,男,教授。E-mail:junhuazhang@nwafu.edu.cn
更新日期/Last Update: 2019-07-10