木聚糖酶组分对杨木酶水解的影响
|
摘要
通过补充木聚糖酶单组分辅助纤维素酶水解稀碱预处理过的杨木，研究木聚糖酶组分对酶解过程中纤维素、半纤维素的转化规律和酶解产物结构的影响。
两种不同的条件下预处理杨木，分别得到底物A（120℃，5%NaOH处理后洗涤，然后80℃，5%NaOH，2%H_2O_2处理）和底物B（80℃，5%NaOH，5%H_2O_2处理）。底物A的纤维素、半纤维素和木质素含量分别为57.8、8.5和18.5%，底物B的纤维素、半纤维素和木质素含量分别为47、9.7和24.8%。
通过凝胶过滤层析分离纯化里氏木聚糖酶得到4种电泳纯组分，分子量分别为98.73×103、21.62×10~3、19.67×10~3和12.85×10~3g/mol，它们分别为木糖苷酶、内切木聚糖酶XYNⅠ、内切木聚糖酶XYNⅡ和外切木聚糖酶。
纤维素酶水解预处理过的杨木72h后，底物A、B的葡萄糖得率分别为74.88和39.2%，木糖得率分别为62.3和57.5%，分别补充添加木糖苷酶、XYNⅠ、XYNⅡ和外切木聚糖酶单组分后，底物A的葡萄糖得率分别为81.8、89.5、89.9和87.2%，提高了7.0、14.62、15.02和12.32个百分点，木糖得率分别为74.5、80.3、79.8和82.4%，提高了12.2、18.0、17.5和20.1个百分点；底物B的葡萄糖得率分别为45.3、62.7、57.8、47.7%，提高了6.1、23.5、18.6和8.5个百分点，木糖得率分别为61.7、70.1、77.1和85.5%，提高了4.2、12.6、19.6和28.0个百分点。可以看出，补充木聚糖酶单组分，可以提高两种底物的葡萄糖和木糖得率，其中XYNⅠ和XYNⅡ能显著提高葡萄糖得率，外切木聚糖酶能显著提高木糖得率；四种木聚糖酶组分对不同组成的底物的作用不同，当底物纤维素含量较低、半纤维素含量较高时，外切木聚糖酶对木糖得率的提高更加明显，说明外切木聚糖酶能更好的降解半纤维素。
在纤维素酶的基础上添加XYNⅠ和XYNⅡ混合组分后，两种底物的葡萄糖得率分别为90.8和54%，提高了16和10个百分点；木糖得率分别为65和81.6%，提高了14和24.2个百分点。这说明，在底物半纤维素含量较高的情况下，XYNⅠ和XYNⅡ组分的复配对半纤维素的降解效果最好。
对添加XYNⅠ和XYNⅡ的酶解液进行凝胶色谱分离，并对其中的低聚木糖组分进行红外光谱、核磁共振~1H NMR和~(13)C NMR分析。可以看出，产物中4-OMe-α-D-GlcpA和(1→4)-β-D-Xylp通过C-O-C键连接，(1→4)-β-D-Xylp与木素通过糖苷键连接形成木素-碳水化合物复合体LCC，(1→4)-β-D-Xylp是通过β-糖苷键连接；由红外可知X2比X1的结构中多了一个亚甲基，由核磁共振图谱可以看出两种产物在结构上有一定区别，具体还有待进一步研究。
This experiment investigated the effects of xylanase components on transformation rulesof cellulose and hemicellulose and the structure of enzyme hydrolyzate during enzymatichydrolysis process via poplar which was pretreated by xylanase single component assistedenzymatic hydrolysis of dilute alkali.
Pretreatment of poplar under two different conditions, substrate A(120℃, treated by5%NaOH, then80℃, treated by5%NaOH and2%H2O2) and substrate B(80℃treated by5%NaOH and5%H2O2) were got respectively. The cellulose, hemicellulose and lignin contents ofsubstrate A and B were57.8,8.5,18.5%and47,9.7,24.8%, respectively.
Four kinds of electrophoresis pure components were got when Richter xylanase wasseparated and edulcorated by gel filtration chromatography. Their molecular weights were98.73×10~3,21.62×10~3,19.67×10~3and2.85×10~3g/mol. They were xylosidase,Endo-xylanase XYNⅠand XYNⅡ, Exo-xylanase.
The glucose yield of substrate A and substrate B were74.88and39.2%and the xyloseyield were62.3and57.5%after pretreated poplar hydrolysed by cellulose for72h. And afteradding xylosidase, Endo-xylanase XYNⅠand XYNⅡ, Exo-xylanase separately, the glucoseyield of substrate A were81.8,89.5,89.9and87.2%, get about7.0,14.62,15.02and12.32percentage point increase, the xylose yield were74.5,80.3,79.8and82.4%, get about12.2,18.0,17.5and20.1percentage point increase; the glucose yield of substrate B were45.3,62.7,57.8and47.7%, get about6.1,23.5,18.6and8.5percentage point increase, the xylose yieldwere61.7,70.1,77.1and85.5%, get about4.2,12.6,19.6and28.0percentage point increase.The results showed that xylanase can increase glucose yield and xylose yield of two substrates.Among them, XYNⅠand XYNⅡcan increase glucose yield obviously, Exo-xylanase canincrease xylose yield obviously. Four xylanase components have different effects on differentsubstrates. When the content of cellulose was low and the hemicellulose was higher,Exo-xylanase improved xylose yield more obviously which indicated that Exo-xylanase candigest hemicellulose better.
The glucose yield of two substrates were90.8and54%, get about16and10percentagepoint increase, the xylose yield were65and81.6%, get about14and24.2percentage pointincrease by adding XYN Ⅰ and XYN Ⅱ mixed-component on the basis of the cellulose. Theresults showed that XYN Ⅰ and XYN Ⅱ mixed-component can digest hemicellulose better in thecase of the substrate with high content of hemicellulose.
The enzymatic hydrolyzate was separated by gel filtration chromatography which wasadded XYN Ⅰ and XYN Ⅱ and xylo-oligosaccharides was analyzed by IR,1H and13C NMR.4-OMe-α-D-GlcpA was connected by C-O-C bond with (1→4)-β-D-Xylp in the products.(1→4)-β-D-Xylp was connected by glycosidic bond with lignin and formed Lignin-CarbohydrateComplexe(LCC).(1→4)-β-D-Xylp was connected by β-glycosidic bond;X2have one-CH2more than X1on the structure by the result of IR. The two products have little difference on thestructure by the result of1H and13C NMR and the specific differences are to be futher studied.
两种不同的条件下预处理杨木，分别得到底物A（120℃，5%NaOH处理后洗涤，然后80℃，5%NaOH，2%H_2O_2处理）和底物B（80℃，5%NaOH，5%H_2O_2处理）。底物A的纤维素、半纤维素和木质素含量分别为57.8、8.5和18.5%，底物B的纤维素、半纤维素和木质素含量分别为47、9.7和24.8%。
通过凝胶过滤层析分离纯化里氏木聚糖酶得到4种电泳纯组分，分子量分别为98.73×103、21.62×10~3、19.67×10~3和12.85×10~3g/mol，它们分别为木糖苷酶、内切木聚糖酶XYNⅠ、内切木聚糖酶XYNⅡ和外切木聚糖酶。
纤维素酶水解预处理过的杨木72h后，底物A、B的葡萄糖得率分别为74.88和39.2%，木糖得率分别为62.3和57.5%，分别补充添加木糖苷酶、XYNⅠ、XYNⅡ和外切木聚糖酶单组分后，底物A的葡萄糖得率分别为81.8、89.5、89.9和87.2%，提高了7.0、14.62、15.02和12.32个百分点，木糖得率分别为74.5、80.3、79.8和82.4%，提高了12.2、18.0、17.5和20.1个百分点；底物B的葡萄糖得率分别为45.3、62.7、57.8、47.7%，提高了6.1、23.5、18.6和8.5个百分点，木糖得率分别为61.7、70.1、77.1和85.5%，提高了4.2、12.6、19.6和28.0个百分点。可以看出，补充木聚糖酶单组分，可以提高两种底物的葡萄糖和木糖得率，其中XYNⅠ和XYNⅡ能显著提高葡萄糖得率，外切木聚糖酶能显著提高木糖得率；四种木聚糖酶组分对不同组成的底物的作用不同，当底物纤维素含量较低、半纤维素含量较高时，外切木聚糖酶对木糖得率的提高更加明显，说明外切木聚糖酶能更好的降解半纤维素。
在纤维素酶的基础上添加XYNⅠ和XYNⅡ混合组分后，两种底物的葡萄糖得率分别为90.8和54%，提高了16和10个百分点；木糖得率分别为65和81.6%，提高了14和24.2个百分点。这说明，在底物半纤维素含量较高的情况下，XYNⅠ和XYNⅡ组分的复配对半纤维素的降解效果最好。
对添加XYNⅠ和XYNⅡ的酶解液进行凝胶色谱分离，并对其中的低聚木糖组分进行红外光谱、核磁共振~1H NMR和~(13)C NMR分析。可以看出，产物中4-OMe-α-D-GlcpA和(1→4)-β-D-Xylp通过C-O-C键连接，(1→4)-β-D-Xylp与木素通过糖苷键连接形成木素-碳水化合物复合体LCC，(1→4)-β-D-Xylp是通过β-糖苷键连接；由红外可知X2比X1的结构中多了一个亚甲基，由核磁共振图谱可以看出两种产物在结构上有一定区别，具体还有待进一步研究。
This experiment investigated the effects of xylanase components on transformation rulesof cellulose and hemicellulose and the structure of enzyme hydrolyzate during enzymatichydrolysis process via poplar which was pretreated by xylanase single component assistedenzymatic hydrolysis of dilute alkali.
Pretreatment of poplar under two different conditions, substrate A(120℃, treated by5%NaOH, then80℃, treated by5%NaOH and2%H2O2) and substrate B(80℃treated by5%NaOH and5%H2O2) were got respectively. The cellulose, hemicellulose and lignin contents ofsubstrate A and B were57.8,8.5,18.5%and47,9.7,24.8%, respectively.
Four kinds of electrophoresis pure components were got when Richter xylanase wasseparated and edulcorated by gel filtration chromatography. Their molecular weights were98.73×10~3,21.62×10~3,19.67×10~3and2.85×10~3g/mol. They were xylosidase,Endo-xylanase XYNⅠand XYNⅡ, Exo-xylanase.
The glucose yield of substrate A and substrate B were74.88and39.2%and the xyloseyield were62.3and57.5%after pretreated poplar hydrolysed by cellulose for72h. And afteradding xylosidase, Endo-xylanase XYNⅠand XYNⅡ, Exo-xylanase separately, the glucoseyield of substrate A were81.8,89.5,89.9and87.2%, get about7.0,14.62,15.02and12.32percentage point increase, the xylose yield were74.5,80.3,79.8and82.4%, get about12.2,18.0,17.5and20.1percentage point increase; the glucose yield of substrate B were45.3,62.7,57.8and47.7%, get about6.1,23.5,18.6and8.5percentage point increase, the xylose yieldwere61.7,70.1,77.1and85.5%, get about4.2,12.6,19.6and28.0percentage point increase.The results showed that xylanase can increase glucose yield and xylose yield of two substrates.Among them, XYNⅠand XYNⅡcan increase glucose yield obviously, Exo-xylanase canincrease xylose yield obviously. Four xylanase components have different effects on differentsubstrates. When the content of cellulose was low and the hemicellulose was higher,Exo-xylanase improved xylose yield more obviously which indicated that Exo-xylanase candigest hemicellulose better.
The glucose yield of two substrates were90.8and54%, get about16and10percentagepoint increase, the xylose yield were65and81.6%, get about14and24.2percentage pointincrease by adding XYN Ⅰ and XYN Ⅱ mixed-component on the basis of the cellulose. Theresults showed that XYN Ⅰ and XYN Ⅱ mixed-component can digest hemicellulose better in thecase of the substrate with high content of hemicellulose.
The enzymatic hydrolyzate was separated by gel filtration chromatography which wasadded XYN Ⅰ and XYN Ⅱ and xylo-oligosaccharides was analyzed by IR,1H and13C NMR.4-OMe-α-D-GlcpA was connected by C-O-C bond with (1→4)-β-D-Xylp in the products.(1→4)-β-D-Xylp was connected by glycosidic bond with lignin and formed Lignin-CarbohydrateComplexe(LCC).(1→4)-β-D-Xylp was connected by β-glycosidic bond;X2have one-CH2more than X1on the structure by the result of IR. The two products have little difference on thestructure by the result of1H and13C NMR and the specific differences are to be futher studied.
引文
[1]顾方媛，陈朝银，石家骥，等.纤维素酶的研究进展与发展趋势[J].微生物学杂志，20o8(0l):83
[2]蒋挺大.木质素[M].北京:化学工业出版社，2001.
[3] Chosdu R, Hilmy N, Erizal Erlinda TB,et al.Radiation and chemical pretreatment of cellulosicwaste[J].Radiat PhysChem,1993,42:695-698.
[4]杨涛.离子液体预处理玉米秸秆及纤维素酶水解研究〔D].天津:天津大学化工学院，2008:
[5] Bedford M R，Partridge G G.EnZymes in Farm Animal Nutrition[M].UK:CABI Publishing，2001.
[6]彭源德.纤维质生物降解制备燃料乙醇研究现状[J].中国麻业科学,2009,31(1):101-103.
[7] Ljungdahl L G, Eriksson K E. Ecology of microbial cellulose degradation [J]. AdvMicrob Ecol,1985,(5):237－299.
[8]王栋,龚大春等.酸法-酶法处理麦秆木质纤维素的工艺研究[J].可再生能源,2008,26(2):50-51.
[9]王沁,赵学慧.黑曲霉(Aspergillus niger)纤维素酶系中内切β－葡聚糖酶性质的研[J].微生物学报,1993,33(6):439–445.
[10] Weimer P J, Hackney J M, French A D. Effects of chemical treatments and heating on the crystallinity ofcelluloses and their implications for evaluating the effect of crystallinity on cellulose biodegradation[J].Biotechnol Bioeng,1995,48:169-178.
[11] Wyman C E. Ethanol from lignocellulosic biomass: technology, economics and opportunities[J].Bioresour Technol,1994,50:3-16.
[12]高洁,汤烈贵.纤维素科学[M].北京：科学出版社,1996.
[13]陈洪章,李佐虎.麦草汽爆作用机理的研究[J].纤维素科学与技术,1999,7(4):14-22.
[14] Grethlein H E, Allen D C, Converse A O. A comparative study of the enzymatic hydrolysis ofacid-pretreated white pine and mixed hardwood[J]. Biotechnol Bioeng,1984,26:1498-1505.
[15]夏延斌.食品化学[M].北京:中国农业出版社,2004:49-50.
[16]刘志明.麦秆表面特性及麦秆刨花板胶接机理的研究[D].东北林业大学博士学位论文,2002:45-46.
[17]连海兰,周定国,尤纪雪.麦秸秆成分剖析及其交合性能的研究[J].林产化学与工业2005,25(3):71.
[18] F an L T, Lee Y, Gharpuray M M. The nature of lignocellulosics and their pretreatments forenzymatic hydrolysis[J]. Adv in Biochem Eng,1982,23:157-187.
[19] Millett M A, Baker A J, Satter L A. Physical and chemical pretreatments for enhancing cellulosesaccharification[J]. Biotechnol Bioeng Symp,1976,6:125-153.
[20]吴赛玉.生物化学[M].安徽:中国科学技术大学出版社,2005:117～118.
[21]咸漠,康亦兼,杨丽.脂肪酶催化反应的研究进展[J].青岛化工学院学报,2000,21(9):189~191.
[22]连惠芗,汪世华.木聚糖酶的研究与应用[J].武汉工业学院学报,2006,25(3):42~45.
[23]薄开静.亲和微滤分离木聚糖酶[D].南京林业大学硕士学位论文,2007.
[24]佘元莉,李秀婷,宋焕禄等.微生物木聚糖酶的研究进展[J].中国酿造,2009,302(2):1-3.
[25]方洛云.木聚糖酶基因的分子生物学与基因工程.livestock and poultry industry.2002(2):2-3
[26] Paice M.G.,Bernier R.Jr.and Jurasek L.Viscosity-enhancing bleaching of hardwood kraft pulp withxylanase from a cloned gene[J]. Biotechnol Bioeng,1988,32(2):235-239.
[27] Clark T A, McDonald A G.,Senior D.Jand Mayers P R.Mananase and xylanase treatment ofsoftwood chemical pulps:effects on pulp properties and bleachability[J]. Biotechnology in Pulp andPaper Manufacture,1990:153-167.
[28]慕娟,问清江,党永,李叶昕,张烁,李文柯,李慧惠.木聚糖酶的开发与应用[J].陕西农业科学,2012,(1):111-115.
[29]刘瑞田,曲音波,杨建云.木聚糖酶分子结构研究进展[J].纤维素科学与技术,1997,5(3):1-6.
[30]陆健.曹钰.陈坚.等.木聚糖酶的产生、性质和应用[J].酿酒,2001,28(6):30-34.
[31]张勤良,王璋,许时婴.中性木聚糖酶在面包制作中的应用[J].食品发酵与工业,2004,30(7):21-23.
[32] Peter Biely. Microbial xylanolytic systems[J]. Trends in Biotechnology,1985,3(11):286-290.
[33] ThomPson N S.Hemicellulose as a biomass resource. Wood and Agricultural ResiduesResearch on Use for Feed, Fuels and Chemicals[M].1983:101-119.
[34]陈洪章.纤维素生物技术[M].北京:化学工业出版社,2005:126-132
[35] Gregg D, Saddler J N. Bioconversion of lignocellulose biomass to et hanol [J]. Biomass and Bioenergy,1995,9(125):287-302.
[36]吴坤,张世敏,朱显峰.木质素生物降解研究进展[J].河南农业大学学报,2000,34(4):349-354.
[37] Charles E. Wyman, Bruce E. Dale, Richard T. Elander. Coordinated development of leading biomasspretreatment technologies [J]. Bioresource Technology,2005(96):1959-1966.
[38] Ye Sun, Jiayang Cheng. Hydrolysis of lignocellosic materials for ethanol prodution: a review[J].Bioresource Technology,2002,83:1-11.
[39]熊犍,叶君.微波对纤维素Ⅰ超分子结构的影响[J].华南理工大学学报,2000,28(3):84-89.
[40]唐爱民,梁文芷.超声波活化处理提高纤维素选择性氧化反应性能的研究[J].声学技术,2000,19(3):121-124.
[41] Yang B., Wyman C E. Effect of xylan and lignin removal by batch and flow through pretreatment on theenzymatic digestibility of corn stover cellulose [J]. Biotechnology and Bioengineering,2004,86(1):88-95.
[42] Garrote G., Dominguez H., Parajo J C. Mild autohydrolysis: an environmentally friendly technology forxylooligosaccharide production from wood [J]. Journal of Chemical Technology and Biotechnology,1999,74(11),1101-1109.
[43]鲁杰,石淑兰,邢效功.NaOH预处理对植物纤维素酶解特性的影响[J].纤维素科学与技术,2004,12(1):1-6.
[44] Chosdu R, Hilmy N, Erizal Erlinda T B, etal. Radiation and chemical pretreatment of cellulosic waste [J].Radiat Phys Chem,1993,42:695-698.
[45]胡民强.臭氧处理粗饲料的研究进展[J].世界农业,2004,1:52-53.
[46] Kim Y S, Newman R H. Solid state13C NMR study of wood degraded by the brown rot fungusGloeophyllum trabeum[J]. Holzforschung,1995,49:109-114.
[47] Bhikhabai R, Johansson G, Petterson G. Isolation of celluloytic enzymes from Trichoderma reesei QM9414[J]. Appl Biochem,1984,(6):336-345
[48] Kiobe J, Kubicek C P. Quantification of the main components of the Trichoderma cellulose complexwith monoclonal antibodies using an enzumelinked immunosorbent assay (ELISA)[J]. Appl. Microbiol.Biotechnology,1990,34:26-30.
[49]隋晓飞，陈嘉川，杨桂花，庞志强.纤维素酶协同木聚糖酶预处理对磨浆能耗及其性能的影响[J].中华纸业，2005，(s):30一33.
[50]涂启梁，付时雨，詹怀宇.纤维素酶和半纤维素酶在制浆造纸工业中的应用[J].西南造纸，2006,35(3):27一29.
[51]庄苏，颜瑞，丁立人，等.纤维素酶与木聚糖酶对象草青贮发酵品质的影响[J].南京农业大学学报，2009，32(4):148一153.
[52]何江川,韩永萍.超滤膜分离法在多糖分离提取中的应用[J].食用菌,2005(1):5-21.
[53]周振.木聚糖酶强化纤维素酶对植物纤维的水解[D].南京林业大学硕士学位论文,2008.
[54] Ohlson G, Tragardh B H. Hagerdel. Enzymatic hydrolysis of sodium-hydroxide-pretreated sallow in anultrafiltration membrane reactor [J]. Biotechnol Bioeng,1984,26:647-653.
[55] Gail Lorenz Miller. Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar[J].Analytical Chemistry,1959,31:426-428.
[56]邬义明主编.植物纤维化学[M].北京：中国轻工业出版社，2000.
[57]刘媛媛,孙君社,裴海生,刘莉,段开红.提高木质纤维素酶水解效率的研究进展[J].中国酿造2011,05:16-20.
[58]于兆海.里氏木酶纤维素酶的分离纯化与酶学性质研究[D].南京林业大学硕士学位论文,2006.
[59] Ogren K, Bura R, Saddler J, etal. Effect of hemicellulose and lignin emoval on enzymatic hydrolysis ofsteam pretreated corn stover[J]. Bioresource Technology,2007,98:2503–2510.
[60]高维.木聚糖酶水解壳聚糖的研究[D].武汉工业学院2008.
[61]江小华．里氏木霉木聚糖酶的分离纯化、酶学特性及降解机理的研究［D］．南京：南京林业大学，2006.
[62] Ghose T K.. Measurement of cellulose activity [J]. Pure＆Appl. Chem.,1987,59(2):257-268.
[63] Alex Berlin, Vera Maximenko, Neil Gilkes.Optimization of Enzyme Complexes for LignocelluloseHydrolysis[J]. Biotechnology and Bioengineering,2007,97(2):288-290.
[64]毛连山,宋向阳,勇强等.里氏木霉木聚糖酶的分离纯化及其性质[J].南京林业大学学报,2002,26(6):14-16.
[65] Cesar T，Mr a V．Purification and properties of the xylanase produced by Thermomyces lanuginosus［J］．Enzyme and Microbial Technology，1996，19(4):289－296．
[66]刘蕾,勇强,余世袁.木聚糖酶分级对高温降解木聚糖酶水解的影响[J].南京林业大学学报,2011,35(3):107-110.
[67]邢玲,江华.木聚糖酶对玉米芯酶水解的影响[J].食品工业科技,2010,31(11):228-231.
[68] Chen, J.L. Studies on the production, purification and characterization of a xylanase from Trichodermasp.1995,185p.(M.S.Thesis,National Chung-Hsing University, Taiwan).
[69]刘家健,陆怡.预处理对纤维素酶降解影响的研究[J],林产化学与工业，1995，15（3）
[70]尉慰奇,武书彬,彭云云.麦草水溶性和碱溶性半纤维素的分离与表征[J].林产化学与工业,2010,30(6):67-70.
[71] A. Ebringerovd, Z. Hromddkov~i, J. Alfiildi.Structural and solution properties of corn cobheteroxylans [J]. Carbohydrate Polymers,1992,19(2):99-105.
[72] Bengtsson, S.; Aman, P. Isolation and chemical characterization of water-soluble arabinoxylans in ryegrain. Carbohydr. Polym.1990,12,267–277.
[73] Tong-Qi Yuan, Feng Xu, Jing He, Run-Cang Sun.Structural and physico-chemical characterization ofhemicelluloses from ultrasound-assisted extractions of partially delignifed fast-growing poplar woodthrough organic solvent and alkaline solutions[J]. Biotechnology Advances,2010,(28),583–593
[74] Feng Peng,Jun-Li Ren,Feng Xu,Jing Bian,Pai Peng,And Ren-Cang Sun. Comparative Study ofHemicelluloses Obtained by Graded Ethanol Precipitation from Sugarcane Bagasse[J]. J.Agriculturaland Food Chemistrty,2009,57,6305–6317
[75]顾瑞军.木素和木素-碳水化合物复合体的化学结构及形成机理[D].华南理工大学硕士论文,2002.
[2]蒋挺大.木质素[M].北京:化学工业出版社，2001.
[3] Chosdu R, Hilmy N, Erizal Erlinda TB,et al.Radiation and chemical pretreatment of cellulosicwaste[J].Radiat PhysChem,1993,42:695-698.
[4]杨涛.离子液体预处理玉米秸秆及纤维素酶水解研究〔D].天津:天津大学化工学院，2008:
[5] Bedford M R，Partridge G G.EnZymes in Farm Animal Nutrition[M].UK:CABI Publishing，2001.
[6]彭源德.纤维质生物降解制备燃料乙醇研究现状[J].中国麻业科学,2009,31(1):101-103.
[7] Ljungdahl L G, Eriksson K E. Ecology of microbial cellulose degradation [J]. AdvMicrob Ecol,1985,(5):237－299.
[8]王栋,龚大春等.酸法-酶法处理麦秆木质纤维素的工艺研究[J].可再生能源,2008,26(2):50-51.
[9]王沁,赵学慧.黑曲霉(Aspergillus niger)纤维素酶系中内切β－葡聚糖酶性质的研[J].微生物学报,1993,33(6):439–445.
[10] Weimer P J, Hackney J M, French A D. Effects of chemical treatments and heating on the crystallinity ofcelluloses and their implications for evaluating the effect of crystallinity on cellulose biodegradation[J].Biotechnol Bioeng,1995,48:169-178.
[11] Wyman C E. Ethanol from lignocellulosic biomass: technology, economics and opportunities[J].Bioresour Technol,1994,50:3-16.
[12]高洁,汤烈贵.纤维素科学[M].北京：科学出版社,1996.
[13]陈洪章,李佐虎.麦草汽爆作用机理的研究[J].纤维素科学与技术,1999,7(4):14-22.
[14] Grethlein H E, Allen D C, Converse A O. A comparative study of the enzymatic hydrolysis ofacid-pretreated white pine and mixed hardwood[J]. Biotechnol Bioeng,1984,26:1498-1505.
[15]夏延斌.食品化学[M].北京:中国农业出版社,2004:49-50.
[16]刘志明.麦秆表面特性及麦秆刨花板胶接机理的研究[D].东北林业大学博士学位论文,2002:45-46.
[17]连海兰,周定国,尤纪雪.麦秸秆成分剖析及其交合性能的研究[J].林产化学与工业2005,25(3):71.
[18] F an L T, Lee Y, Gharpuray M M. The nature of lignocellulosics and their pretreatments forenzymatic hydrolysis[J]. Adv in Biochem Eng,1982,23:157-187.
[19] Millett M A, Baker A J, Satter L A. Physical and chemical pretreatments for enhancing cellulosesaccharification[J]. Biotechnol Bioeng Symp,1976,6:125-153.
[20]吴赛玉.生物化学[M].安徽:中国科学技术大学出版社,2005:117～118.
[21]咸漠,康亦兼,杨丽.脂肪酶催化反应的研究进展[J].青岛化工学院学报,2000,21(9):189~191.
[22]连惠芗,汪世华.木聚糖酶的研究与应用[J].武汉工业学院学报,2006,25(3):42~45.
[23]薄开静.亲和微滤分离木聚糖酶[D].南京林业大学硕士学位论文,2007.
[24]佘元莉,李秀婷,宋焕禄等.微生物木聚糖酶的研究进展[J].中国酿造,2009,302(2):1-3.
[25]方洛云.木聚糖酶基因的分子生物学与基因工程.livestock and poultry industry.2002(2):2-3
[26] Paice M.G.,Bernier R.Jr.and Jurasek L.Viscosity-enhancing bleaching of hardwood kraft pulp withxylanase from a cloned gene[J]. Biotechnol Bioeng,1988,32(2):235-239.
[27] Clark T A, McDonald A G.,Senior D.Jand Mayers P R.Mananase and xylanase treatment ofsoftwood chemical pulps:effects on pulp properties and bleachability[J]. Biotechnology in Pulp andPaper Manufacture,1990:153-167.
[28]慕娟,问清江,党永,李叶昕,张烁,李文柯,李慧惠.木聚糖酶的开发与应用[J].陕西农业科学,2012,(1):111-115.
[29]刘瑞田,曲音波,杨建云.木聚糖酶分子结构研究进展[J].纤维素科学与技术,1997,5(3):1-6.
[30]陆健.曹钰.陈坚.等.木聚糖酶的产生、性质和应用[J].酿酒,2001,28(6):30-34.
[31]张勤良,王璋,许时婴.中性木聚糖酶在面包制作中的应用[J].食品发酵与工业,2004,30(7):21-23.
[32] Peter Biely. Microbial xylanolytic systems[J]. Trends in Biotechnology,1985,3(11):286-290.
[33] ThomPson N S.Hemicellulose as a biomass resource. Wood and Agricultural ResiduesResearch on Use for Feed, Fuels and Chemicals[M].1983:101-119.
[34]陈洪章.纤维素生物技术[M].北京:化学工业出版社,2005:126-132
[35] Gregg D, Saddler J N. Bioconversion of lignocellulose biomass to et hanol [J]. Biomass and Bioenergy,1995,9(125):287-302.
[36]吴坤,张世敏,朱显峰.木质素生物降解研究进展[J].河南农业大学学报,2000,34(4):349-354.
[37] Charles E. Wyman, Bruce E. Dale, Richard T. Elander. Coordinated development of leading biomasspretreatment technologies [J]. Bioresource Technology,2005(96):1959-1966.
[38] Ye Sun, Jiayang Cheng. Hydrolysis of lignocellosic materials for ethanol prodution: a review[J].Bioresource Technology,2002,83:1-11.
[39]熊犍,叶君.微波对纤维素Ⅰ超分子结构的影响[J].华南理工大学学报,2000,28(3):84-89.
[40]唐爱民,梁文芷.超声波活化处理提高纤维素选择性氧化反应性能的研究[J].声学技术,2000,19(3):121-124.
[41] Yang B., Wyman C E. Effect of xylan and lignin removal by batch and flow through pretreatment on theenzymatic digestibility of corn stover cellulose [J]. Biotechnology and Bioengineering,2004,86(1):88-95.
[42] Garrote G., Dominguez H., Parajo J C. Mild autohydrolysis: an environmentally friendly technology forxylooligosaccharide production from wood [J]. Journal of Chemical Technology and Biotechnology,1999,74(11),1101-1109.
[43]鲁杰,石淑兰,邢效功.NaOH预处理对植物纤维素酶解特性的影响[J].纤维素科学与技术,2004,12(1):1-6.
[44] Chosdu R, Hilmy N, Erizal Erlinda T B, etal. Radiation and chemical pretreatment of cellulosic waste [J].Radiat Phys Chem,1993,42:695-698.
[45]胡民强.臭氧处理粗饲料的研究进展[J].世界农业,2004,1:52-53.
[46] Kim Y S, Newman R H. Solid state13C NMR study of wood degraded by the brown rot fungusGloeophyllum trabeum[J]. Holzforschung,1995,49:109-114.
[47] Bhikhabai R, Johansson G, Petterson G. Isolation of celluloytic enzymes from Trichoderma reesei QM9414[J]. Appl Biochem,1984,(6):336-345
[48] Kiobe J, Kubicek C P. Quantification of the main components of the Trichoderma cellulose complexwith monoclonal antibodies using an enzumelinked immunosorbent assay (ELISA)[J]. Appl. Microbiol.Biotechnology,1990,34:26-30.
[49]隋晓飞，陈嘉川，杨桂花，庞志强.纤维素酶协同木聚糖酶预处理对磨浆能耗及其性能的影响[J].中华纸业，2005，(s):30一33.
[50]涂启梁，付时雨，詹怀宇.纤维素酶和半纤维素酶在制浆造纸工业中的应用[J].西南造纸，2006,35(3):27一29.
[51]庄苏，颜瑞，丁立人，等.纤维素酶与木聚糖酶对象草青贮发酵品质的影响[J].南京农业大学学报，2009，32(4):148一153.
[52]何江川,韩永萍.超滤膜分离法在多糖分离提取中的应用[J].食用菌,2005(1):5-21.
[53]周振.木聚糖酶强化纤维素酶对植物纤维的水解[D].南京林业大学硕士学位论文,2008.
[54] Ohlson G, Tragardh B H. Hagerdel. Enzymatic hydrolysis of sodium-hydroxide-pretreated sallow in anultrafiltration membrane reactor [J]. Biotechnol Bioeng,1984,26:647-653.
[55] Gail Lorenz Miller. Use of Dinitrosalicylic Acid Reagent for Determination of Reducing Sugar[J].Analytical Chemistry,1959,31:426-428.
[56]邬义明主编.植物纤维化学[M].北京：中国轻工业出版社，2000.
[57]刘媛媛,孙君社,裴海生,刘莉,段开红.提高木质纤维素酶水解效率的研究进展[J].中国酿造2011,05:16-20.
[58]于兆海.里氏木酶纤维素酶的分离纯化与酶学性质研究[D].南京林业大学硕士学位论文,2006.
[59] Ogren K, Bura R, Saddler J, etal. Effect of hemicellulose and lignin emoval on enzymatic hydrolysis ofsteam pretreated corn stover[J]. Bioresource Technology,2007,98:2503–2510.
[60]高维.木聚糖酶水解壳聚糖的研究[D].武汉工业学院2008.
[61]江小华．里氏木霉木聚糖酶的分离纯化、酶学特性及降解机理的研究［D］．南京：南京林业大学，2006.
[62] Ghose T K.. Measurement of cellulose activity [J]. Pure＆Appl. Chem.,1987,59(2):257-268.
[63] Alex Berlin, Vera Maximenko, Neil Gilkes.Optimization of Enzyme Complexes for LignocelluloseHydrolysis[J]. Biotechnology and Bioengineering,2007,97(2):288-290.
[64]毛连山,宋向阳,勇强等.里氏木霉木聚糖酶的分离纯化及其性质[J].南京林业大学学报,2002,26(6):14-16.
[65] Cesar T，Mr a V．Purification and properties of the xylanase produced by Thermomyces lanuginosus［J］．Enzyme and Microbial Technology，1996，19(4):289－296．
[66]刘蕾,勇强,余世袁.木聚糖酶分级对高温降解木聚糖酶水解的影响[J].南京林业大学学报,2011,35(3):107-110.
[67]邢玲,江华.木聚糖酶对玉米芯酶水解的影响[J].食品工业科技,2010,31(11):228-231.
[68] Chen, J.L. Studies on the production, purification and characterization of a xylanase from Trichodermasp.1995,185p.(M.S.Thesis,National Chung-Hsing University, Taiwan).
[69]刘家健,陆怡.预处理对纤维素酶降解影响的研究[J],林产化学与工业，1995，15（3）
[70]尉慰奇,武书彬,彭云云.麦草水溶性和碱溶性半纤维素的分离与表征[J].林产化学与工业,2010,30(6):67-70.
[71] A. Ebringerovd, Z. Hromddkov~i, J. Alfiildi.Structural and solution properties of corn cobheteroxylans [J]. Carbohydrate Polymers,1992,19(2):99-105.
[72] Bengtsson, S.; Aman, P. Isolation and chemical characterization of water-soluble arabinoxylans in ryegrain. Carbohydr. Polym.1990,12,267–277.
[73] Tong-Qi Yuan, Feng Xu, Jing He, Run-Cang Sun.Structural and physico-chemical characterization ofhemicelluloses from ultrasound-assisted extractions of partially delignifed fast-growing poplar woodthrough organic solvent and alkaline solutions[J]. Biotechnology Advances,2010,(28),583–593
[74] Feng Peng,Jun-Li Ren,Feng Xu,Jing Bian,Pai Peng,And Ren-Cang Sun. Comparative Study ofHemicelluloses Obtained by Graded Ethanol Precipitation from Sugarcane Bagasse[J]. J.Agriculturaland Food Chemistrty,2009,57,6305–6317
[75]顾瑞军.木素和木素-碳水化合物复合体的化学结构及形成机理[D].华南理工大学硕士论文,2002.