Efficient enzymatic hydrolysis of high-solid lignocellulosic substrate with formulated cellulase preparation at low enzyme loading

doi:10.16085/j.issn.1000-6613.2024-1538

Abstract

Abstract:

An efficient enzymatic hydrolysis method for pretreatment of bagasse with alkali-catalyzed glycerol aqueous solution was established. Single factor and orthogonal experiments determined that the above substrates were enzymolized in batches (50g/L) under the action of surfactants (15mg/g Triton X-100, 25mg/g PEG 6000, 25mg/g AEO-9) and auxiliary enzymes (2.5mg/g xylanase). The enzymatic hydrolysis rate of 6FPU/g dry substrate was 88.6% after 72h. The enzymolysis efficiency of the substrate was the highest, with the yield of glucose and xylose being 78.8% and 76.4% at 72h, respectively, and the concentration of total fermentable sugar was 140g/L. It could be seen that the strategy proposed in this study had obvious industrial application potential in terms of cellulase loading capacity (6FPU/g), enzymatic hydrolysis period (72h), glucose/xylose yield (about 80%) and fermentable sugar concentration.

Key words: lignocellulosic biomass, surfactant, enzymatic saccharification, fermentable sugar

摘要：

建立了一种碱催化甘油水溶液预处理蔗渣的高效酶解糖化方法。单因素和正交实验确定上述基质在表面活性剂（15mg/g Triton X-100、25mg/g PEG 6000、25mg/g AEO-9）和辅助酶（2.5mg/g木聚糖酶）作用下批次酶解（50g/L，6FPU/g干底物）72h后酶解率为88.6%；所获得的底物酶解效率最高，72h的葡萄糖得率达到78.8%，木糖得率为76.4%，总可发酵性糖浓度为140g/L。可见，本研究所提出的底物浓醪酶解糖化策略在纤维素酶加载量（6FPU/g）、酶解周期（72h）、葡萄糖/木糖得率（约80%）和可发酵性糖浓度等方面具有明显工业化应用潜力。

关键词: 木质纤维素生物质, 表面活性剂, 酶解糖化, 可发酵性糖

CLC Number:

TQ352.78

BIAN Mingyuan, WANG Chen, LIU Zicheng, LIU Qiangqiang, SUN Chihe, REN Hongyan, SUN Fubao. Efficient enzymatic hydrolysis of high-solid lignocellulosic substrate with formulated cellulase preparation at low enzyme loading[J]. Chemical Industry and Engineering Progress, 2025, 44(11): 6688-6694.

边明源, 王晨, 刘子成, 刘强强, 孙驰贺, 任洪艳, 孙付保. 复配定制纤维素酶实现浓醪底物的低酶高效水解糖化[J]. 化工进展, 2025, 44(11): 6688-6694.

Figures/Tables 7

References 16

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序号	Triton X-100/mg·g^-1	PEG 6000/mg·g^-1	AEO-9/mg·g^-1	EX/mg·g^-1	48h酶解率/%	72h酶解率/%
1	5	7	25	1.5	76.9	84.1
2	5	15	35	2.5	78.3	84.4
3	5	25	45	3.5	78.3	82.4
4	5	35	65	4.5	78.3	79.3
5	10	7	35	3.5	76.9	82.4
6	10	15	25	4.5	76.9	83.4
7	10	25	65	1.5	78.3	79.7
8	10	35	45	2.5	78.3	82.4
9	15	7	45	4.5	67.8	74.6
10	15	15	65	3.5	75.6	82.4
11	15	25	25	2.5	84.5	88.6
12	15	35	35	1.5	76.9	81.4
13	25	7	65	2.5	78.3	83.7
14	25	15	45	1.5	70.2	76.3
15	25	25	35	4.5	78.3	82.4
16	25	35	25	3.5	76.9	84.4

序号	Triton X-100/mg·g^-1	PEG 6000/mg·g^-1	AEO-9/mg·g^-1	EX/mg·g^-1	48h酶解率/%	72h酶解率/%
1	5	7	25	1.5	76.9	84.1
2	5	15	35	2.5	78.3	84.4
3	5	25	45	3.5	78.3	82.4
4	5	35	65	4.5	78.3	79.3
5	10	7	35	3.5	76.9	82.4
6	10	15	25	4.5	76.9	83.4
7	10	25	65	1.5	78.3	79.7
8	10	35	45	2.5	78.3	82.4
9	15	7	45	4.5	67.8	74.6
10	15	15	65	3.5	75.6	82.4
11	15	25	25	2.5	84.5	88.6
12	15	35	35	1.5	76.9	81.4
13	25	7	65	2.5	78.3	83.7
14	25	15	45	1.5	70.2	76.3
15	25	25	35	4.5	78.3	82.4
16	25	35	25	3.5	76.9	84.4

时间/h	生产速率/g·L^-1·h^-1
时间/h	5%	8%	10%	12%	14%
3	10.40	8.33	8.25	8.29	—
4	3.39	7.20	5.42	2.82	—
6	2.37	1.69	1.53	1.98	—
9	3.62	2.54	2.94	2.26	12.11
12	1.36	1.69	1.36	1.51	1.29
15	1.58	1.77	1.58	1.79	1.86
18	1.47	1.20	1.69	1.13	0.97
24	3.28	2.54	2.03	1.69	1.94
36	0.48	0.65	0.61	0.66	0.60
72	0.37	0.58	0.55	0.54	0.27

时间/h	生产速率/g·L^-1·h^-1
时间/h	5%	8%	10%	12%	14%
3	10.40	8.33	8.25	8.29	—
4	3.39	7.20	5.42	2.82	—
6	2.37	1.69	1.53	1.98	—
9	3.62	2.54	2.94	2.26	12.11
12	1.36	1.69	1.36	1.51	1.29
15	1.58	1.77	1.58	1.79	1.86
18	1.47	1.20	1.69	1.13	0.97
24	3.28	2.54	2.03	1.69	1.94
36	0.48	0.65	0.61	0.66	0.60
72	0.37	0.58	0.55	0.54	0.27

模式	补料方式/g·L^-1									酶解率/%
模式	1h	2h	3h	6h	9h	12h	18h	24h	30h	48h	72h
1	40	40	40	—	—	—	—	—	—	61.9	74.6
2	—	—	—	40	—	40	40	—	—	69.5	78.8
3	—	—	—	40	—	—	40	40	—	63.6	75.4
4	—	—	—	—	—	—	40	40	40	60.2	72.0
对照	—	—	—	—	—	—	—	—	—	58.5	68.6