木聚糖酶的生产及应用研究
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摘要
本论文主要优化了木霉RS30(Trichoderma RS30)生产木聚糖酶的培养条件。并完成了木聚糖酶与纤维素酶对玉米秸秆的降解研究及含有木聚糖酶的复合酶影响蛋鸡生产性能的应用研究。
通过优化试验获得产木聚糖酶的最佳条件是：温度为28℃、培养基pH值为5.0、种龄为96h、接种量1×107个孢子、摇床转速150r／min、培养时间144h。
通过木聚糖酶和纤维素酶对玉米秸秆的降解研究,确定的酶解条件与效果为：在酶解体系pH4.5、温度50℃、固液比为1：15、摇床转速为100r／min、木聚糖酶120U／mL、纤维素酶60u／mL时,降解24h,玉米秸秆降解率为12.2%。
蛋鸡饲料中添加复合酶制剂没有显著改变鸡蛋品质,使用了0.2%的复合酶制剂能有效提高蛋鸡的生产性能,平均产蛋率提高了2.53%,产蛋量提高了3.56%,料蛋比下降6.36%,盈利率提高了16.23%。
In this study, the producing condition of xylanase by Trichoderma RS30 has been optimized. Then the xylanase was used for hydrolysis of corn straw cooperating with cellulase. An enzyme mixture containing xylanase was added into chick feed. The effect of enzyme mixture on eggs producing has also been investigated in this study.
The optimal conditions of xylanase producing are as following:28℃, pH 5.0,96 hour-old inoculum, inoculum size of 1×107 spore, shake speed of 150 r/min, and 144 hours of fermentation.
In the study on corn straw enzymatic hydrolysis, the performance of xylanase and cellulase mixture was evaluated. The highest degradation rate (12.2%) of corn straw was achieved under the conditions as following:pH 4.5,50℃, a solid to liquid ratio of 1:15, shake speed of 100 r/min, dosage of 120 U/mL xylanase and 60 u/mL cellulase,24 hours of hydrolysis.
The enzyme mixture added into feeds has no significant effects on quality of eggs. The results indicated that addition of 0.2% complex enzymes in feed effectively improved the eggs production performance with 2.53% increase of average eggs producing rate,3.56% increase of eggs production,6.36% decrease of feed to egg ratio, and 16.23% increase of profile.
通过优化试验获得产木聚糖酶的最佳条件是：温度为28℃、培养基pH值为5.0、种龄为96h、接种量1×107个孢子、摇床转速150r／min、培养时间144h。
通过木聚糖酶和纤维素酶对玉米秸秆的降解研究,确定的酶解条件与效果为：在酶解体系pH4.5、温度50℃、固液比为1：15、摇床转速为100r／min、木聚糖酶120U／mL、纤维素酶60u／mL时,降解24h,玉米秸秆降解率为12.2%。
蛋鸡饲料中添加复合酶制剂没有显著改变鸡蛋品质,使用了0.2%的复合酶制剂能有效提高蛋鸡的生产性能,平均产蛋率提高了2.53%,产蛋量提高了3.56%,料蛋比下降6.36%,盈利率提高了16.23%。
In this study, the producing condition of xylanase by Trichoderma RS30 has been optimized. Then the xylanase was used for hydrolysis of corn straw cooperating with cellulase. An enzyme mixture containing xylanase was added into chick feed. The effect of enzyme mixture on eggs producing has also been investigated in this study.
The optimal conditions of xylanase producing are as following:28℃, pH 5.0,96 hour-old inoculum, inoculum size of 1×107 spore, shake speed of 150 r/min, and 144 hours of fermentation.
In the study on corn straw enzymatic hydrolysis, the performance of xylanase and cellulase mixture was evaluated. The highest degradation rate (12.2%) of corn straw was achieved under the conditions as following:pH 4.5,50℃, a solid to liquid ratio of 1:15, shake speed of 100 r/min, dosage of 120 U/mL xylanase and 60 u/mL cellulase,24 hours of hydrolysis.
The enzyme mixture added into feeds has no significant effects on quality of eggs. The results indicated that addition of 0.2% complex enzymes in feed effectively improved the eggs production performance with 2.53% increase of average eggs producing rate,3.56% increase of eggs production,6.36% decrease of feed to egg ratio, and 16.23% increase of profile.
引文
[1]陈洪章.纤维素生物技术[M].北京：化学工业出版社,现代生物技术与医药科技出版中心,2005.
[2]Joseleau J. P., Comtat J., Ruel K. Chemical structure of xylans and their interaction in the plant cell walls. In:Progress in biotechnology 7.Xylans and Xylanases[M](Visser J., Beldman G., Kusters-van Someren M. A., et al. Eds). Elsevier, Amserdam.1992,1-16.
[3]崔建涛.树脂固定化木聚糖酶及其动力学研究[D].河南：河南农业大学,2007.
[4]Coughlan M.P., Hazlewood G.P.1,4-D-xylan-degrading enzyme system:Biochemistry, molecular biology and plications [J]. Biotechnology and Applied Biochemistry, 1993,V17:259-289.
[5]胡沂淮,邵蔚蓝.木聚糖酶[J].生命的化学,2002,22(3)：281.
[6]张红莲,姚斌,范云六.木聚糖酶的分子生物学及其应用[J].生物技术通报,2002(3)：23.
[7]Okazaki W.,Akiba T.,Horikoshi K., et al. Purification and characterization of xylanase from Flavus[J]. Joural of Fermentation Technology,1985, V62:361-369.
[8]Nakanishi K.,Arai H.,YasuiT. Purification and some properties of xylanase from Cryptococcus[J]. Canadian Journal of Microbiology,1984, V28:733-739.
[9]Keskar S.S., Srinivasan M.C., Deshpande V.V. Chemical modification of a xylanase from a thermotolerant Streptomyces T7[J].Biochemical Journal,1989, V261:49-51.
[10]Vrsanska M., Gorbacheva I.V., Kratky Z., et al. Reaction pathway of substrate degradation by an acidic endo-1,4-β-xylanase of A.niger[J].Biochemica Biophysica Acta, 1982,704(1):114-122.
[11]Kubackova M., Karacsony I.S., Toman R. Purification of xylanases from wood rooting fungus Trametes hirsute [J]. Folia Microbiologia 1976, V21:28-35.
[12]Frederick M.M., Chin-Hen K., Fredericks J.R., et al.Purification and characterization of endo-xylanases from A.niger two isoenzyme active on xylan backbone near branch point[J]. Biotechnology and Bioengineering 1985, V27:527-532.
[13]Paice M.G., Jurasek L., Carpenter M.R., et al. Production, charaterization and partical amino acid sequence of xylanase from Schizophyllum commune [J]. Applied and Environmental Microbiology,1978, V36:802-808.
[14]Breccia J.D., Castro G.R., Baigori M.D., et al. Screening of xylanolytic bacteria using a colour plate method[J]. Journal of Apllied Bacteriology,1995, V78:469-472.
[15]包怿红,木聚糖酶工程菌的构建[M].黑龙江：东北林业大学出版社,2005.
[16]郑建仙.功能性食品甜味剂[M].北京：中国轻工业出版社,1995.
[17]陈瑞娟.新型低聚糖的介绍[J].食品与发酵工业,1993,V19(2)：82-90.
[18]许正宏,熊筱晶,陶文沂.低聚木糖的生产及应用研究进展[J].食品与发酵工业,2001,V28(1)：56-59.
[19]杨瑞金,许时婴,王璋.低聚木糖的酶法生产[J].无锡轻工大学学报,2000,V19(4)：340-344.
[20]王海,李里特.用玉米芯酶法制备低聚木糖[J].食品科学,2002,V23(5)：81-83.
[21]张艳艳,薛文通,汪正强等.卷须链木聚糖酶水解玉米芯汽爆液及酵母发酵精致低聚木糖[J].食品与发酵工业,2004,V30(4)：12-16.
[22]薛业敏,毛忠贵,邵蔚蓝.利用玉米芯木聚糖酶法制备低聚木糖的研究[J].中国酿造,2003,(6)：7-9.
[23]郭夏丽,刘新育,陈红歌.微生物酶法生产低聚木糖条件的研究[J].食品与发酵工业,2002,V29(9)：57-59.
[24]Godfrey T., West S. Industrial Enzymology (Second Edition) [M]. New York:Stockton Press,1996.
[25]Rombouts F. M. Enzymes in fruit and vegetable juice technology[J]. Process Biochemistry,1978, V13:9-13.
[26]Andorsson R., Westerlund E., Aman P. Natural variations in the contents of structural elements of water-extractable non-starch polysaccharides in white flour[J]. Journal of Cereal Science,1994, V19(1):77-82.
[27]Izydorczyk M.S., Biliaderis C.G. Cereal Arabinoxylans:Advances in Structure and Physicochemical Properties[J]. Carbohydrate Polymers,1995, V28:33-48.
[28]邓伟、李秀婷、江正强,等.橄榄绿链霉菌木聚糖酶组对面包品质的改善,2005,V20(1)：1-5.
[29]Courtin C.M., Delcour J.A. Arabinoxylans and Endoxylanases in Wheat Flour Bread-making[J]. Journal of Cereal Science,2002, V35(3):225-243.
[30]Gan Z., Ellis P.R., Schofield J.D. Mini review:gas cell stabilisation and gas retention in wheat bread dough[J]. Journal of Cereal Science,1995, V21(3):215-230.
[31]Ogasawara H., Takahashi K, Iitsuka K, et al. Contribution of Hemicellulase in Shochu koji to the Resolution of Barley in the Shochu Mash[J]. Journal of the Brewing Society of Japan,1991, V86(4):304-307.
[32]陆健,曹钰,陈坚,等.木聚糖酶的产生、性质和应用[J].酿酒,2001,V28(6)：30-34.
[33]Bhat M.K. Cellulases and related enzymes in biotechnology[J]. Biotechnology Advances, 2000,V18:355-383.
[34]国家排放新标准 全面提升造纸行业污染控制水平.中国轻工业网.http://paper.clii.com.cn/Criterion/infoshow.asp?showid=5245,2008-7-25.
[35]Kulkarni N.,Shendye A., Rao M. Molecular and biotechnological aspects of xylanase[J]. FEMS Microbiology Review,1999, V23(4):411-456.
[36]罗贵民,曹淑桂,张今.酶工程[M].北京：化学工业出版社,现代生物技术与医药科技出版中心,2002.
[37]韩正康.家禽及猪营养中的酶制剂[A].韩正康饲料酶制剂国际学术研讨会论文集[C],1996,3-6.
[38]刘强.我国麦类饲料中非淀粉多糖抗营养作用机理的研究[D].北京：中国农业科学院,1998.
[39]Ratty D., Gollnisch K., Schulz E., et al. The effect of an antimicrobial feed additive, an on-starch-polysaccharide (NSP)-hydrolyzing enzyme and their combination on the overall digestibility of nutrients and the content of metabolizable energy in growing pigs[J]. Landbouforschung Volkenrode,1998, V48(4):169-175,34.
[40]Ikegami S., Tsuchihashi F., Harada H., et al. Effect of viscous indigestible polysaccharides on pancreatic biliary secretion and digestive organs in rats [J]. Journal of Nutrition,1990, V120(4):353-360.
[41]郝波,王辉.生物饲料在21世纪的前景[J].中国饲料,2000,V2：19.
[42]Bedford M.R. The effect of enzymes on digestion [J]. Journal of Applied Poultry Research,1996, V5:370-378.
[43]Annison G. Commercial enzyme supplementation of wheat-based diets raises ileal glucanase activities and improves apparent metabolizable energy, starch and pentosan digestibilities in broiler chickens [J]. Animal Feed Science and Technology,1992, V38:105-121.
[44]Classen H.L., Bedford M.R. The use of enzymes to improve the nutritive value of poultry feeds [A]. Recent Advances in Animal Nutrition[C], Butterworth-Heinemann Ltd., Oxford, 1991,95-116.
[45]张雷.中国能源供应战略的调整[J].中国能源,1999,V3：12-14.
[46]曲音波.纤维素乙醇产业化[J].化学进展,2007,V19(7)：1098-1108.
[47]Jeewon L. Biological coversion of lignocellulosic biomass to ethanol. Journal Biotechnology,1997, V56(1):1-24.
[48]周振.木聚糖酶强化纤维素酶对植物纤维的水解[D].南京：南京林业大学,2008.
[49]DB61／T465-2009,陕西省地方标准-木聚糖酶制剂[S].陕西：陕西省质量技术监督局,2009.
[50]QB 2583-2003,纤维素酶制剂[S].北京：中华人民共和国发展改革委员会,2003.
[51]温天舜,周大强,刘一勋.随机数据分析与处理-应用概率统计.开封：河南大学出版社,1993.
[52]张晓梅,任发政,葛克山.饲料中添加桑饲料对蛋鸡生产性能和鸡蛋品质的影响[J].食品科学,2007,V28(3)：89-91.
[2]Joseleau J. P., Comtat J., Ruel K. Chemical structure of xylans and their interaction in the plant cell walls. In:Progress in biotechnology 7.Xylans and Xylanases[M](Visser J., Beldman G., Kusters-van Someren M. A., et al. Eds). Elsevier, Amserdam.1992,1-16.
[3]崔建涛.树脂固定化木聚糖酶及其动力学研究[D].河南：河南农业大学,2007.
[4]Coughlan M.P., Hazlewood G.P.1,4-D-xylan-degrading enzyme system:Biochemistry, molecular biology and plications [J]. Biotechnology and Applied Biochemistry, 1993,V17:259-289.
[5]胡沂淮,邵蔚蓝.木聚糖酶[J].生命的化学,2002,22(3)：281.
[6]张红莲,姚斌,范云六.木聚糖酶的分子生物学及其应用[J].生物技术通报,2002(3)：23.
[7]Okazaki W.,Akiba T.,Horikoshi K., et al. Purification and characterization of xylanase from Flavus[J]. Joural of Fermentation Technology,1985, V62:361-369.
[8]Nakanishi K.,Arai H.,YasuiT. Purification and some properties of xylanase from Cryptococcus[J]. Canadian Journal of Microbiology,1984, V28:733-739.
[9]Keskar S.S., Srinivasan M.C., Deshpande V.V. Chemical modification of a xylanase from a thermotolerant Streptomyces T7[J].Biochemical Journal,1989, V261:49-51.
[10]Vrsanska M., Gorbacheva I.V., Kratky Z., et al. Reaction pathway of substrate degradation by an acidic endo-1,4-β-xylanase of A.niger[J].Biochemica Biophysica Acta, 1982,704(1):114-122.
[11]Kubackova M., Karacsony I.S., Toman R. Purification of xylanases from wood rooting fungus Trametes hirsute [J]. Folia Microbiologia 1976, V21:28-35.
[12]Frederick M.M., Chin-Hen K., Fredericks J.R., et al.Purification and characterization of endo-xylanases from A.niger two isoenzyme active on xylan backbone near branch point[J]. Biotechnology and Bioengineering 1985, V27:527-532.
[13]Paice M.G., Jurasek L., Carpenter M.R., et al. Production, charaterization and partical amino acid sequence of xylanase from Schizophyllum commune [J]. Applied and Environmental Microbiology,1978, V36:802-808.
[14]Breccia J.D., Castro G.R., Baigori M.D., et al. Screening of xylanolytic bacteria using a colour plate method[J]. Journal of Apllied Bacteriology,1995, V78:469-472.
[15]包怿红,木聚糖酶工程菌的构建[M].黑龙江：东北林业大学出版社,2005.
[16]郑建仙.功能性食品甜味剂[M].北京：中国轻工业出版社,1995.
[17]陈瑞娟.新型低聚糖的介绍[J].食品与发酵工业,1993,V19(2)：82-90.
[18]许正宏,熊筱晶,陶文沂.低聚木糖的生产及应用研究进展[J].食品与发酵工业,2001,V28(1)：56-59.
[19]杨瑞金,许时婴,王璋.低聚木糖的酶法生产[J].无锡轻工大学学报,2000,V19(4)：340-344.
[20]王海,李里特.用玉米芯酶法制备低聚木糖[J].食品科学,2002,V23(5)：81-83.
[21]张艳艳,薛文通,汪正强等.卷须链木聚糖酶水解玉米芯汽爆液及酵母发酵精致低聚木糖[J].食品与发酵工业,2004,V30(4)：12-16.
[22]薛业敏,毛忠贵,邵蔚蓝.利用玉米芯木聚糖酶法制备低聚木糖的研究[J].中国酿造,2003,(6)：7-9.
[23]郭夏丽,刘新育,陈红歌.微生物酶法生产低聚木糖条件的研究[J].食品与发酵工业,2002,V29(9)：57-59.
[24]Godfrey T., West S. Industrial Enzymology (Second Edition) [M]. New York:Stockton Press,1996.
[25]Rombouts F. M. Enzymes in fruit and vegetable juice technology[J]. Process Biochemistry,1978, V13:9-13.
[26]Andorsson R., Westerlund E., Aman P. Natural variations in the contents of structural elements of water-extractable non-starch polysaccharides in white flour[J]. Journal of Cereal Science,1994, V19(1):77-82.
[27]Izydorczyk M.S., Biliaderis C.G. Cereal Arabinoxylans:Advances in Structure and Physicochemical Properties[J]. Carbohydrate Polymers,1995, V28:33-48.
[28]邓伟、李秀婷、江正强,等.橄榄绿链霉菌木聚糖酶组对面包品质的改善,2005,V20(1)：1-5.
[29]Courtin C.M., Delcour J.A. Arabinoxylans and Endoxylanases in Wheat Flour Bread-making[J]. Journal of Cereal Science,2002, V35(3):225-243.
[30]Gan Z., Ellis P.R., Schofield J.D. Mini review:gas cell stabilisation and gas retention in wheat bread dough[J]. Journal of Cereal Science,1995, V21(3):215-230.
[31]Ogasawara H., Takahashi K, Iitsuka K, et al. Contribution of Hemicellulase in Shochu koji to the Resolution of Barley in the Shochu Mash[J]. Journal of the Brewing Society of Japan,1991, V86(4):304-307.
[32]陆健,曹钰,陈坚,等.木聚糖酶的产生、性质和应用[J].酿酒,2001,V28(6)：30-34.
[33]Bhat M.K. Cellulases and related enzymes in biotechnology[J]. Biotechnology Advances, 2000,V18:355-383.
[34]国家排放新标准 全面提升造纸行业污染控制水平.中国轻工业网.http://paper.clii.com.cn/Criterion/infoshow.asp?showid=5245,2008-7-25.
[35]Kulkarni N.,Shendye A., Rao M. Molecular and biotechnological aspects of xylanase[J]. FEMS Microbiology Review,1999, V23(4):411-456.
[36]罗贵民,曹淑桂,张今.酶工程[M].北京：化学工业出版社,现代生物技术与医药科技出版中心,2002.
[37]韩正康.家禽及猪营养中的酶制剂[A].韩正康饲料酶制剂国际学术研讨会论文集[C],1996,3-6.
[38]刘强.我国麦类饲料中非淀粉多糖抗营养作用机理的研究[D].北京：中国农业科学院,1998.
[39]Ratty D., Gollnisch K., Schulz E., et al. The effect of an antimicrobial feed additive, an on-starch-polysaccharide (NSP)-hydrolyzing enzyme and their combination on the overall digestibility of nutrients and the content of metabolizable energy in growing pigs[J]. Landbouforschung Volkenrode,1998, V48(4):169-175,34.
[40]Ikegami S., Tsuchihashi F., Harada H., et al. Effect of viscous indigestible polysaccharides on pancreatic biliary secretion and digestive organs in rats [J]. Journal of Nutrition,1990, V120(4):353-360.
[41]郝波,王辉.生物饲料在21世纪的前景[J].中国饲料,2000,V2：19.
[42]Bedford M.R. The effect of enzymes on digestion [J]. Journal of Applied Poultry Research,1996, V5:370-378.
[43]Annison G. Commercial enzyme supplementation of wheat-based diets raises ileal glucanase activities and improves apparent metabolizable energy, starch and pentosan digestibilities in broiler chickens [J]. Animal Feed Science and Technology,1992, V38:105-121.
[44]Classen H.L., Bedford M.R. The use of enzymes to improve the nutritive value of poultry feeds [A]. Recent Advances in Animal Nutrition[C], Butterworth-Heinemann Ltd., Oxford, 1991,95-116.
[45]张雷.中国能源供应战略的调整[J].中国能源,1999,V3：12-14.
[46]曲音波.纤维素乙醇产业化[J].化学进展,2007,V19(7)：1098-1108.
[47]Jeewon L. Biological coversion of lignocellulosic biomass to ethanol. Journal Biotechnology,1997, V56(1):1-24.
[48]周振.木聚糖酶强化纤维素酶对植物纤维的水解[D].南京：南京林业大学,2008.
[49]DB61／T465-2009,陕西省地方标准-木聚糖酶制剂[S].陕西：陕西省质量技术监督局,2009.
[50]QB 2583-2003,纤维素酶制剂[S].北京：中华人民共和国发展改革委员会,2003.
[51]温天舜,周大强,刘一勋.随机数据分析与处理-应用概率统计.开封：河南大学出版社,1993.
[52]张晓梅,任发政,葛克山.饲料中添加桑饲料对蛋鸡生产性能和鸡蛋品质的影响[J].食品科学,2007,V28(3)：89-91.
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