碱-羟甲基化预处理对甘蔗渣酶解和发酵效率的影响

doi:10.3969/j.issn.0253-2417.2020.04.012

摘要/Abstract

摘要：

以甘蔗渣（SCB）为原料，经1.4%氢氧化钠处理制得碱处理甘蔗渣（AP-SCB），再经羟甲基化反应制备碱-羟甲基化预处理甘蔗渣（AHP-SCB），考察甲醛用量对提高甘蔗渣酶水解效率和乙醇发酵得率的影响。实验结果表明：在甘蔗渣碱-羟甲基预处理过程中，当甘蔗渣与甲醛质量比为1：1.5，预处理2 h后，木质素脱除率为55.68%；在纤维素酶用量为20 FPIU/g（以葡聚糖质量计）时，72 h酶水解得率为89.61%，较未处理甘蔗渣（水解得率9.09%）提高了8.86倍；酶水解液经24 h发酵后，乙醇质量浓度从10.93 g/L（AP-SCB）增加至12.96 g/L（AHP-SCB-1.5）。通过红外光谱分析预处理前后物料结构的变化，并进行疏水度和酶可及度的测定，AHP-SCB-1.5的疏水度（0.127 L/g）和酶可及度（505.73 mg/g）分别是AP-SCB的64.64%和1.11倍；碱和羟甲基化反应在预处理中起到协同作用，改善甘蔗渣的亲水性和酶可及性。

关键词: 羟甲基化, 碱处理, 甘蔗渣, 酶解, 发酵

Abstract:

To investigate the effects of formaldehyde dosage on improving enzymatic hydrolysis efficiency and ethanol fermentation yield of sugarcane bagasse, the sequential NaOH and hydroxymethylation pretreatment (AHP) were used to treat sugarcane bagasse (SCB) with different proportion of formaldehyde by hydroxymethylation reaction. The results showed that when SCB was treated by AHP with substrate and formaldehyde ratio of 1:1.5 for 2 h, the delignification rate was 55.68% and the mass fraction of cellulose was 67.20%.Under the cellulase dosage of 20 FPIU/g(based on the mass of glucan), the 72 h glucose yield was 89.61%, which was 8.86 times higher than that of untreated SCB (9.09%). After 24 h fermentation, the ethanol concentration increased from 10.93 g/L of AP-SCB to 12.96 g/L of AHP-SCB-1.5.The structural features of the pretreated sugarcane bagasse were measured by FT-IR, and the hydrophobicity and cellulase accessibility were studied. The hydrophobicity (0.127 L/g) and enzymatic accessibility (505.73 mg/g) of AHP-SCB-1.5 were 64.64% and 1.11 times of AP-SCB, respectively. The results showed that alkali and hydroxymethylation had a synergistic effect on pretreatment process, which could improve the hydrophilicity and cellulase accessibility of materials.

Key words: hydroxymethylation, alkali pretreatment, sugarcane bagasse, enzymatic hydrolysis, fermentation

中图分类号:

TQ35

金艳,杨海艳,史正军,王大伟,徐高峰,杨静. 碱-羟甲基化预处理对甘蔗渣酶解和发酵效率的影响[J]. 林产化学与工业, 2020, 40(4): 86-92.

Yan JIN,Haiyan YANG,Zhengjun SHI,Dawei WANG,Gaofeng XU,Jing YANG. Effect of Alkali-hydroxymethylation Pretreatment on Sugarcane Bagasse Enzymatic Hydrolysis and Fermentation Efficiency[J]. Chemistry and Industry of Forest Products, 2020, 40(4): 86-92.

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参考文献 25

1	REVELL L E , BODEKER G E , HUCK P E , et al. Impacts of the production and consumption of biofuels on stratospheric ozone[J]. Geophysical Research Letters, 2012, 39 (10): 10804- 10809.
2	VALLEJOS , MARÍA EVANGELINA , ZAMBON M D , et al. Low liquid-solid ratio (LSR) hot water pretreatment of sugarcane bagasse[J]. Green Chemistry, 2012, 14 (7): 1982- 1989.
3	吕晓静.基于碱的木质纤维素预处理和酶解及相关机理研究[D].广州:暨南大学, 2018.
	LV X J, Alkali-based pretreatment and enzymatic hydrolysis of lignocellulosic biomass and their related mechanisms[D]. Guangzhou: Jinan University, 2018.
4	HUANG C , HE J , MIN D , et al. Understanding the nonproductive enzyme adsorption and physicochemical properties of residual lignins in moso bamboo pretreated with sulfuric acid and kraft pulping[J]. Applied Biochemistry & Biotechnology, 2016, 180 (8): 1508- 1523.
5	LU X , WANG C , LI X , et al. Studying nonproductive adsorption ability and binding approach of cellobiohydrolase to lignin during bioconversion of lignocellulose[J]. Energy & Fuels, 2017, 31 (12): 14393- 14400.
6	LOU H , WANG M , LAI H , et al. Reducing non-productive adsorption of cellulase and enhancing enzymatic hydrolysis of lignocelluloses by noncovalent modification of lignin with lignosulfonate[J]. Bioresource Technology, 2013, 146 (10): 478- 484.
7	WANG Z , ZHU J , FU Y , et al. Lignosulfonate-mediated cellulase adsorption:Enhanced enzymatic saccharification of lignocellulose through weakening nonproductive binding to lignin[J]. Biotechnology for Biofuels, 2013, 6 (1): 156- 167.
8	TSEGAYE B , BALOMAJUMDER C , ROY P . Alkali delignification and Bacillus sp.BMP01 hydrolysis of rice straw for enhancing biofuel yields[J]. 2019, 43 (1): 136- 146.
9	WENJUN Y , ZHENGJUN S , HAIYAN Y , et al. Effect of alkaline lignin modification on cellulase-lignin interactions and enzymatic saccharification yield[J]. Biotechnology for Biofuels, 2018, 11 (1): 214- 227.
10	QIN C , CLARKE K , LI K . Interactive forces between lignin and cellulase as determined by atomic force microscopy[J]. Biotechnology for Biofuels, 2014, 7 (1): 65- 74.
11	汪骏, 金永灿. 木质素模型物羟甲基化反应的机理研究[J]. 纤维素科学与技术, 2011, 19 (2): 1- 9.
	WANG J , JIN Y C . Effect of NaOH concentration in pretreatment on rice rice straw hydrolysis inhibitor's compounds[J]. Journal of Cellulose Science and Technology, 2011, 19 (2): 1- 9.
12	LI X , LI M , PU Y , et al. Inhibitory effects of lignin on enzymatic hydrolysis:The role of lignin chemistry and molecular weight[J]. Renewable energy, 2018, 123, 664- 674.
13	YANG J , TU M , XIA C , et al. Effect of fenton pretreatment on C₁ and C₆ oxidation of cellulose and its enzymatic hydrolyzability[J]. ACS Sustainable Chemistry & Engineering, 2019, 7 (7): 7071- 7079.
14	应文俊, 吴凯, 史正军, 等. 磷酸联合过氧化氢预处理玉米芯及其酶水解效率的影响[J]. 林产化学与工业, 2018, 38 (5): 100- 106.
	YING W J , WU K , SHI Z J , et al. Pretreatment of corn cob by the combination of H₃PO₄-H₂O₂ and their effect on enzymatic hydrolysis efficiency[J]. Chemistry and Industry of Forest Products, 2018, 38 (5): 100- 106.
15	YANG H , SHI Z , XU G , et al. Bioethanol production from bamboo with alkali-catalyzed liquid hot water pretreatment[J]. Bioresource Technology, 2019, 274, 261- 290.
16	李志强, 费本华, 江泽慧. 发酵抑制物对葡萄糖发酵产乙醇的影响[J]. 化工进展, 2015, 34 (S1): 80- 84.
	LI Z Q , FEI B H , JIANG Z H . Effect of fermentation inhibitors on ethanol production of glucose fermentation[J]. Chemical Industry and Engineering Progress, 2015, 34 (S1): 80- 84.
17	WU X , HUANG C , ZHAI S , et al. Improving enzymatic hydrolysis efficiency of wheat straw through sequential autohydrolysis and alkaline post-extraction[J]. Bioresource Technology, 2017, 251, 374- 380.
18	GESSNER A , WAICZ R , LIESKE A , et al. Nanoparticles with decreasing surface hydrophobicities:Influence on plasma protein adsorption[J]. International Journal of Pharmaceutics, 2000, 196 (2): 245- 249.
19	INGLESBY M K , ZERONIAN S H . Direct dyes as molecular sensors to characterize cellulose substrates[J]. Cellulose, 2002, 9 (1): 19- 29.
20	陆青山.木质纤维素对里氏木霉产纤维素酶的诱导[D].南京:南京林业大学, 2016.
	LU Q S.Induction effect of lignocellulose to cellulase synthesisby Trichoderma reesei[D]. Nanjing: Nanjing Forestry University, 2016.
21	HE J , HUANG C , LAI C , et al. Relations between moso bamboo surface properties pretreated by Kraft cooking and dilute acid with enzymatic digestibility[J]. Applied biochemistry and biotechnology, 2017, 183 (4): 1- 13.
22	YANG J , ZHANG X , YONG Q , et al. Three-stage enzymatic hydrolysis of steam-exploded corn stover at high substrate concentration[J]. Bioresource Technology, 2011, 102 (7): 4905- 4908.
23	YU H , YOU Y , LEI F , et al. Comparative study of alkaline hydrogen peroxide and organosolv pretreatments of sugarcane bagasse to improve the overall sugar yield[J]. Bioresource Technology, 2015, 187, 161- 166.
24	CANILHA L , CHANDEL A K , THAIS S D S M , et al. Bioconversion of sugarcane biomass into ethanol:An overview about composition, pretreatment methods, detoxification of hydrolysates, enzymatic saccharification, and ethanol fermentation[J]. Journal of Biomedicine & Biotechnology, 2012, 2012, 1- 15.
25	丁驰, 黄慧琴, 张树栋, 等. 不同浓度NaOH预处理对稻秸水解液中抑制物组分的影响[J]. 食品与发酵工业, 2015, 41 (9): 51- 56.
	DING C , HUANG H Q , ZHANG S D , et al. Effect of NaOH concentrations in pretreatment on rice straw hydrolysis inhibitor's compounds[J]. Food and Fermentation Industries, 2015, 41 (9): 51- 56.

样品¹⁾ sample	m(甘蔗渣):m(甲醛) m(SCB):m(HCHO)	葡聚糖/% glucan	木聚糖/% xylan	木质素/% lignin	木质素脱除率/% delignification
SCB		46.88±0.34	22.73±1.12	22.52±1.13
AP-SCB		66.83±1.34	23.31±1.17	11.52±0.57	48.85±2.34
AHP-SCB-1.5	1:1.5	67.20±1.36	22.67±1.13	9.98±0.50	55.68±0.98
AHP-SCB-3	1:3	68.10±1.41	23.05±1.15	9.12±0.46	59.50±0.67
AHP-SCB-5	1:5	69.50±1.48	23.31±1.17	8.76±0.44	61.10±2.16
HP-SCB-1.5	1:1.5	50.46±0.53	23.14±0.98	21.76±0.34	3.37±1.32
HP-SCB-3	1:3	53.36±0.45	22.86±1.19	20.68±0.59	8.17±1.54
HP-SCB-5	1:5	52.13±0.78	23.40±1.02	20.91±0.32	7.15±1.57

样品 sample	酶可及度/(mg·g^-1) enzyme accessibility	乙醇质量浓度/(g·L^-1) mass concentration of ethanol	乙醇得率/% ethanol yield
SCB	155.02±1.22
AP-SCB	455.61±1.13	10.93±0.54	74.71±1.13
AHP-SCB-1.5	505.73±1.04	12.96±0.35	82.27±1.05
AHP-SCB-3	502.26±2.47	11.44±0.57	80.39±2.47
AHP-SCB-5	467.51±1.58	9.28±0.46	64.76±1.58
HP-SCB-1.5	202.48±3.56	1.12±0.11	68.54±1.24
HP-SCB-3	202.46±2.47	1.14±0.21	65.13±0.76
HP-SCB-5	219.29±0.98	1.07±0.17	66.13±1.26

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