Effect of pretreatment using ammonia water combined with freezing and thawing on enzymatic hydrolysis and sugar production of sorghum straw
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摘要:目的
降低预处理成本、提高秸秆预处理后的酶解效果，模拟自然界低温环境并结合氨水对高粱秸秆进行预处理。
方法通过单因素试验分别探究氨水低温冻融预处理中浸泡液的液固质量比、冷冻温度、冷冻时长、氨水质量分数对高粱秸秆酶解的影响，采用正交试验对预处理条件进行优化，对预处理前后高粱秸秆的成分采用范式法测定，物理化学结构用红外光谱和X射线衍射分析。
结果单因素试验中，浸泡液的液固质量比、冷冻温度、冷冻时长和氨水质量分数在不同水平下均显著提高了高粱秸秆酶解还原糖的产量(P<0.05)。正交试验最优预处理条件为浸泡液的液固质量比12，冷冻时长12 h，冷冻温度−10 ℃，氨水质量分数8%。相较于未进行预处理的秸秆，氨水低温冻融处理的秸秆半纤维素含量下降42.42%；木质素含量下降50.76%；秸秆的还原糖产量为302.87 mg·g−1，较未预处理组提高了80.34%；纤维素结晶度提高 了57.02%。
结论氨水低温冻融预处理能有效破坏高粱秸秆木质纤维素间原有的连接结构，溶解半纤维素，木质素的单体和聚合结构被破坏，提高了高粱秸秆的酶解还原糖得率以及纤维素结晶度。
Abstract:ObjectiveTo reduce the cost of pretreatment and improve the enzymatic hydrolysis effect afterstraw pretreatment, sorghum straw was pretreated with simulated natural low temperature environment andammonia water.
MethodWe studied the effects of the liquid-solid ratio in soaking solution, freezingtemperature, freezing time and ammonia content in pretreatment using ammonia water combined withfreezing and thawing on enzymatic hydrolysis of sorghum straw through single factor tests. We optimized the pretreatment conditions using orthogonal test design. The compositions of sorghum straw before and after pretreatment were measured using normal form method, and the physical and chemical structures were investigated using infrared spectrum and X-ray diffraction analyses.
ResultIn single factor tests, liquid-solid ratio in soaking solution, freezing temperature, freezing time and ammonia content at different levels all significantly increased the production of reducing sugar through enzymatic digestion(P<0.05). The optimum pretreatment conditions of the orthogonal test were 12 liquid-solid ratio in soaking solution, 12 h freezing time, 10 ℃ freezing temperature, and ammonia content of 8%. Compared with straw without pretreatment, in straw with pretreatment using ammonia water combined with freezing and thawing, the hemicellulose content decreased by 42.42% , the lignin content decreased by 50.76%, the yield of reducing sugar for straw was 302.87 mg·g−1, which was 80.34% higher than that of straw without pretreatment, and the crystallinity of cellulose increased by 57.02%.
ConclusionThe pretreatment using ammonia water combined with freezing and thawing can effectively destroy the original connection structure between lignocellulose of sorghum straw, dissolve hemicellulose, destroy the monomer and polymeric structure of lignin. It improves the yield of reducing sugar by enzymatic hydrolysis of sorghum straw, and also improves the crystallinity of sorghum straw cellulose.
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Keywords:
- sorghum straw /
- freezing and thawing /
- ammonia water /
- pretreatment /
- enzymatic hydrolysis
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图 1 不同单因素对高粱秸秆酶解还原糖产量的影响
“*”表示与CK差异显著(P<0.05，t检验)
Figure 1. Effect of different single factor on the yield of reducing sugar by enzymatic hydrolysis of sorghum straw
“*” indicates significant difference from CK (P<0.05, t test)
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图 2 高粱秸秆氨水低温冻融联合预处理前后傅里叶红外光谱图
Figure 2. Fourier infrared spectra of sorghum straw before and after pretreatment using ammonia water combined with freezing and thawing
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图 3 高粱秸秆氨水低温冻融联合预处理前后X射线衍射图谱
Figure 3. X-ray diffraction pattern of sorghum straw before and after pretreatment using ammonia water combined with freezing and thawing
表 1 高粱秸秆氨水低温冻融联合预处理正交试验因素与水平
Table 1 Orthogonal test factors and levels of pretreatments of sorghum straw using ammonia water combined with freezing and thawing
水平
Level液固质量比
Liquid-solid ratio冷冻时长/h
Freezing time冷冻温度/℃
Freezing temperature氨水质量分数/%
Ammonia content1 8∶1 4.0 −10 4 2 10∶1 12.0 −15 6 3 12∶1 24.0 −20 8 
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表 2 高粱秸秆氨水低温冻融联合预处理正交表及分析结果
Table 2 Orthogonal table and analysis result for pretreatment of sorghum straw using ammonia water combined with freezing and thawing
试验号
Test No.因素及水平 Factor and level 还原糖产量/(mg·g−1)
Yield of reducing sugar液固质量比
Liquid-solid ratio冷冻时长
Freezing time冷冻温度
Freezing temperture氨水质量分数
Ammonia content1(CK) 167.94 2 1 1 1 1 235.11 3 1 2 2 2 235.04 4 1 3 3 3 248.54 5 2 1 2 3 291.45 6 2 2 3 1 224.36 7 2 3 1 2 243.72 8 3 1 3 2 248.49 9 3 2 1 3 302.87 10 3 3 2 1 251.93 K1 239.56 258.35 260.57 237.13 K2 253.18 254.09 259.47 242.42 K3 267.76 248.06 240.47 280.95 R 28.20 10.29 20.10 43.82
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表 3 氨水低温冻融联合预处理对高粱秸秆成分变化的影响
Table 3 Influence of pretreatment using ammonia water combined with freezing and thawing on compositional change of sorghum straw
w/% 秸秆 Straw 纤维素 Cellulose 半纤维素 Hemicellulose 木质素 Lignin 灰分 Ash 未处理 Untreated 42.23±1.78 28.57±0.14 14.46±1.75 0.97±0.02 试验最优 Best in test 38.83±1.56 16.45±0.58 7.12±1.82 0.99±0.01
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