黑曲霉改性玉米秸秆制备溢油吸附剂的研究--本科毕业论文.docx
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黑曲霉改性玉米秸秆制备溢油吸附剂的研究--本科毕业论文,摘要玉米秸秆作为天然的溢油吸附剂，其吸油量为4.9-6.2 g原油/g溢油吸附剂，可通过真菌改性提高其吸油量。作者以玉米秸秆作为黑曲霉固体发酵培养基的基质，利用黑曲霉在生长过程中产生的菌丝机械穿插作用，增加玉米秸秆的孔隙，同时黑曲霉代谢分泌木质纤维素酶，由内向外降解木质纤维素，使比表面积增大，达到对玉米秸秆改性的效果。...
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摘 要
玉米秸秆作为天然的溢油吸附剂，其吸油量为4.9-6.2 g原油/g溢油吸附剂，可通过真菌改性提高其吸油量。作者以玉米秸秆作为黑曲霉固体发酵培养基的基质，利用黑曲霉在生长过程中产生的菌丝机械穿插作用，增加玉米秸秆的孔隙，同时黑曲霉代谢分泌木质纤维素酶，由内向外降解木质纤维素，使比表面积增大，达到对玉米秸秆改性的效果。
作者对黑曲霉改性玉米秸秆制备溢油吸附剂的反应条件（温度、反应时间、含水率和菌体投加量）及机理进行了初步的探讨。
结果表明：通过L25(56)4因素5水平的正交实验得出，在无外加碳源的情况下，黑曲霉改性玉米秸秆制备吸附剂的条件的最优组合是：菌体投加量为2 %、干料：水为1：3，反应温度为25℃，反应时间为12d；影响程度为反应时间>反应温度>(干料：水)>菌液投加量。在菌体投加量为2 %，干料：水为1：3的条件下，于30℃培养9d，吸油量达到最大值，是玉米原材料的2.8倍。在外加碳源（蔗糖），投加量为2%的情况下，最佳的反应时间为6d，最佳含水率为1：3、最佳温度为35℃，其最大吸油量是玉米秸秆原材料的2.9倍。外加碳源使吸油量最大值到来时间提前，吸油量增大。研究表明当酶活作用在一定时间和一定量的情况下，能使溢油吸附剂结构孔隙率较高且持油性强，达到吸油量的最大值。BET、XRD、扫描电镜和红外光谱的表征结果分别显示经过黑曲霉改性后溢油吸附剂比表面积增大，结晶度上升，孔隙增多，但表面集团的种类没有发生变化。
关键字：黑曲霉；玉米秸秆；溢油吸附剂；吸油量；酶活
Abstract
As natural sorbents in oil spill cleanup, the amount of oil sorbed by the corn stalk is
4.9-6.2 g crude oil/g sorbents, which can be improved by fungus modification. The author make corn stalk as the substrate of Aspergillus niger solid fermentation medium, taking advantage of the enzymes for lignocellulose degration secreted during Aspergillus niger‘s metabolism . Åt the same time, Aspergillus niger can reach the effect of modifying corn stalks because they are filamentous and have the ability to produce prolific spores to increase the specific surface area.
Preliminary study on the optimization (temperature, reaction time, water ratio and biomass dosage) and the mechanism of corn stalks modified by Aspergillus niger to make oil spill cleanup sorbents have been finished by author.
The results showed that: In the L25(56) orthogonal test(4 factors and 5 level) ,without external carbon source, the optimal combination of Aspergillus niger modifing corn stalks to make oil spill cleanup sorbents is: 12d reaction time, 25 ℃ reaction temperature , drier: water= 1:3(water ratio), 2% biomass dosage. The level of influence is time> the reaction temperature > water ratio > bacterium dosage. In the condition of 2% biomass dosage , drier:water= 1:3(water ratio), 30 ℃ and the reaction time is 9d, oil absorption reached maximum,which is 2.8 times than the raw corn stalk. With external carbon source, the optimization is 6d reaction time, 35 ℃ reaction temperature, drier: water= 1:3(water ratio) with 2% biomass dosage . The amount of oil sorbed by the oil spill cleanup sorbents is 2.9 times than the raw corn stalks.The study have showed that external carbon source can make the maximum of oil absorptivity not only came ealier, but also higher .Study shows that the enzymes for lignocellulose degration militate in a certain amount for certain reaction time can make the oil spill cleanup sorbents have higher multiaperture oil absorbent structure and the stronger ability to hold oil. In this way ,the amount of oill sorbed reach a satisfied value.The result of BET, XRD, SEM and FTIR spectra characterization show that the oil spill cleanup sorbents modified by Aspergillus niger have
larger surface area ,bigger crystallinity and more pores than the raw corn stalk, separately. However, the chemical grounps of the sorbents’ surface are unchanged.
Keyword: Aspergillus niger, corn stalk, oil spill cleanup sorbents, the amount of oil sorbed, enzyme activity
目 录
摘 要 I
Abstract II
第一章 绪论 1
1.1溢油吸附剂 1
1.2溢油吸附剂的研究现状 1
1.3真菌改性天然溢油吸附剂 4
1.3 .1玉米秸秆结构及木质纤维素组成及特点 4
1.3.2生物酶 6
1.3.3固态发酵 8
1.3.4真菌改性天然吸附材料的机理与特点 8
1.4研究目的及意义 9
1.4.1研究目的 9
1.4.2研究意义 9
1.5主要研究内容 10
第二章 材料与方法 11
2.1 实验材料 11
2.1.1供试菌种 11
2.1.2供试玉米秸秆 11
2.1.3供试原油 11
2.1.4培养基 11
2.1.5 固态发酵营养液 13
2.1.6主要仪器及设备 13
2.1.7主要试剂 14
2.1.8试剂配制 16
2.2研究内容与方法 18
2.2.1无外加碳源情况下，黑曲霉改性玉米秸秆的条件研究 18
2.2.2外加碳源情况下，黑曲霉改性玉米秸秆的条件研究 20
2.2.3黑曲霉改性玉米秸秆的步骤 20
2.2.4木质纤维素酶的酶活变化的测定 21
2.2.5木质纤维素含量的测定 23
2.2.6吸油实验步骤 23
2.2.7溢油吸附剂的表征方法 24
第三章 结果与讨论 26
3.1标准曲线的绘制 26
3.1.1葡萄糖标准曲线（DNS光度法）的绘制 26
3.1.2木糖标准曲线（地衣酚光度法）的绘制 26
3.1.3葡萄糖标准曲线（蒽酮光度法）的绘制 27
3.2黑曲霉改性前玉米秸秆吸油情况 28
3.3无外加碳源情况下，黑曲霉改性玉米秸秆的条件研究分析 28
3.3.1无外加碳源情况..
玉米秸秆作为天然的溢油吸附剂，其吸油量为4.9-6.2 g原油/g溢油吸附剂，可通过真菌改性提高其吸油量。作者以玉米秸秆作为黑曲霉固体发酵培养基的基质，利用黑曲霉在生长过程中产生的菌丝机械穿插作用，增加玉米秸秆的孔隙，同时黑曲霉代谢分泌木质纤维素酶，由内向外降解木质纤维素，使比表面积增大，达到对玉米秸秆改性的效果。
作者对黑曲霉改性玉米秸秆制备溢油吸附剂的反应条件（温度、反应时间、含水率和菌体投加量）及机理进行了初步的探讨。
结果表明：通过L25(56)4因素5水平的正交实验得出，在无外加碳源的情况下，黑曲霉改性玉米秸秆制备吸附剂的条件的最优组合是：菌体投加量为2 %、干料：水为1：3，反应温度为25℃，反应时间为12d；影响程度为反应时间>反应温度>(干料：水)>菌液投加量。在菌体投加量为2 %，干料：水为1：3的条件下，于30℃培养9d，吸油量达到最大值，是玉米原材料的2.8倍。在外加碳源（蔗糖），投加量为2%的情况下，最佳的反应时间为6d，最佳含水率为1：3、最佳温度为35℃，其最大吸油量是玉米秸秆原材料的2.9倍。外加碳源使吸油量最大值到来时间提前，吸油量增大。研究表明当酶活作用在一定时间和一定量的情况下，能使溢油吸附剂结构孔隙率较高且持油性强，达到吸油量的最大值。BET、XRD、扫描电镜和红外光谱的表征结果分别显示经过黑曲霉改性后溢油吸附剂比表面积增大，结晶度上升，孔隙增多，但表面集团的种类没有发生变化。
关键字：黑曲霉；玉米秸秆；溢油吸附剂；吸油量；酶活
Abstract
As natural sorbents in oil spill cleanup, the amount of oil sorbed by the corn stalk is
4.9-6.2 g crude oil/g sorbents, which can be improved by fungus modification. The author make corn stalk as the substrate of Aspergillus niger solid fermentation medium, taking advantage of the enzymes for lignocellulose degration secreted during Aspergillus niger‘s metabolism . Åt the same time, Aspergillus niger can reach the effect of modifying corn stalks because they are filamentous and have the ability to produce prolific spores to increase the specific surface area.
Preliminary study on the optimization (temperature, reaction time, water ratio and biomass dosage) and the mechanism of corn stalks modified by Aspergillus niger to make oil spill cleanup sorbents have been finished by author.
The results showed that: In the L25(56) orthogonal test(4 factors and 5 level) ,without external carbon source, the optimal combination of Aspergillus niger modifing corn stalks to make oil spill cleanup sorbents is: 12d reaction time, 25 ℃ reaction temperature , drier: water= 1:3(water ratio), 2% biomass dosage. The level of influence is time> the reaction temperature > water ratio > bacterium dosage. In the condition of 2% biomass dosage , drier:water= 1:3(water ratio), 30 ℃ and the reaction time is 9d, oil absorption reached maximum,which is 2.8 times than the raw corn stalk. With external carbon source, the optimization is 6d reaction time, 35 ℃ reaction temperature, drier: water= 1:3(water ratio) with 2% biomass dosage . The amount of oil sorbed by the oil spill cleanup sorbents is 2.9 times than the raw corn stalks.The study have showed that external carbon source can make the maximum of oil absorptivity not only came ealier, but also higher .Study shows that the enzymes for lignocellulose degration militate in a certain amount for certain reaction time can make the oil spill cleanup sorbents have higher multiaperture oil absorbent structure and the stronger ability to hold oil. In this way ,the amount of oill sorbed reach a satisfied value.The result of BET, XRD, SEM and FTIR spectra characterization show that the oil spill cleanup sorbents modified by Aspergillus niger have
larger surface area ,bigger crystallinity and more pores than the raw corn stalk, separately. However, the chemical grounps of the sorbents’ surface are unchanged.
Keyword: Aspergillus niger, corn stalk, oil spill cleanup sorbents, the amount of oil sorbed, enzyme activity
目 录
摘 要 I
Abstract II
第一章 绪论 1
1.1溢油吸附剂 1
1.2溢油吸附剂的研究现状 1
1.3真菌改性天然溢油吸附剂 4
1.3 .1玉米秸秆结构及木质纤维素组成及特点 4
1.3.2生物酶 6
1.3.3固态发酵 8
1.3.4真菌改性天然吸附材料的机理与特点 8
1.4研究目的及意义 9
1.4.1研究目的 9
1.4.2研究意义 9
1.5主要研究内容 10
第二章 材料与方法 11
2.1 实验材料 11
2.1.1供试菌种 11
2.1.2供试玉米秸秆 11
2.1.3供试原油 11
2.1.4培养基 11
2.1.5 固态发酵营养液 13
2.1.6主要仪器及设备 13
2.1.7主要试剂 14
2.1.8试剂配制 16
2.2研究内容与方法 18
2.2.1无外加碳源情况下，黑曲霉改性玉米秸秆的条件研究 18
2.2.2外加碳源情况下，黑曲霉改性玉米秸秆的条件研究 20
2.2.3黑曲霉改性玉米秸秆的步骤 20
2.2.4木质纤维素酶的酶活变化的测定 21
2.2.5木质纤维素含量的测定 23
2.2.6吸油实验步骤 23
2.2.7溢油吸附剂的表征方法 24
第三章 结果与讨论 26
3.1标准曲线的绘制 26
3.1.1葡萄糖标准曲线（DNS光度法）的绘制 26
3.1.2木糖标准曲线（地衣酚光度法）的绘制 26
3.1.3葡萄糖标准曲线（蒽酮光度法）的绘制 27
3.2黑曲霉改性前玉米秸秆吸油情况 28
3.3无外加碳源情况下，黑曲霉改性玉米秸秆的条件研究分析 28
3.3.1无外加碳源情况..
