糙皮侧耳(Pleurotus ostreatus)615对小麦秸秆木质纤维素降解效果的研究
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摘要
本试验利用糙皮侧耳(Pleurotus ostreatus)615菌株固体发酵小麦秸秆,旨在研究糙皮侧耳615菌株对小麦秸秆木质纤维素的降解效果。液体培养糙皮侧耳615菌株并测定1~10 d产漆酶、锰过氧化物酶活力变化;随后利用糙皮侧耳615菌株种子液固体发酵经1.00%Ca O处理的小麦秸秆,25℃发酵30 d,测定小麦秸秆中性洗涤纤维、酸性洗涤纤维、纤维素、半纤维素、木质素以及粗蛋白质含量变化;最后对发酵后的小麦秸秆进行电镜扫描,观察其结构变化。结果表明:(1)漆酶活力从第1天至第4天不断上升,并在第4天达到最大值1428.00 U/L,从第5天开始下降,且在第10天下降到399.00 U/L;锰过氧化物酶活力从第1天至第5天不断上升,并在第5天达到最大值15.45 U/L,从第6天开始下降,且在第10天下降到4.74 U/L。(2)小麦秸秆经糙皮侧耳615菌株发酵后中性洗涤纤维含量显著降低(P<0.05),且比对照组降低了4.23%,酸性洗涤纤维的含量显著升高(P<0.05),且比对照组升高了4.19%;纤维素含量极显著高于对照组(P<0.01),且比对照组高了37.17%,半纤维素、木质素的含量显著降低(P<0.01),分别比对照组降低了19.87%、43.75%;同时,粗蛋白质的含量显著提高(P<0.01),比对照组提高了4.14%。(3)扫描电镜结果显示,小麦秸秆经糙皮侧耳615菌株发酵后,其粗纤维可被有效降解。液体培养糙皮侧耳615菌株在第4天、第5天产漆酶、锰过氧化物酶能力较强;小麦秸秆经糙皮侧耳615菌株固体发酵30 d后,小麦秸秆中的木质素含量、半纤维素含量均显著降低,粗蛋白质含量显著升高。
In this study,the fungus Pleurotus ostreatus 615 strain was fermented by solid-state fermentation of wheat straw,designed to study effects of Pleurotus ostreatus 615 strain on fiber degradation in wheat straw.Liquid culture of Pleurotus ostreatus 615 strain and the activity of laccase and manganese peroxidase in 1 ~ 10 days were determined.Then the wheat straw which treaded with 1.00% Ca O were fermented at 25 ℃ for 30 days. And the contents of neutral detergent fiber,acid detergent fiber,cellulose,hemicelluloses,lignin and crude protein of wheat straw were determined by solid fermentation of Pleurotus ostreatus 615 strain seed liquid.Last,the fermented wheat straw was scanned by the electron microscope to observe its structural changes.The results showed as follows:(1)Laccase activity increased from first day to fourth day and reached a maximum of 1428.00 U/L on fourth day,decreased from fifth day and creased to399.00 U/L on tenth day;manganese peroxidase activity increased from first day to fifth day and reached a maximum of 15.45 U/L on fifth day,decreased from sixth day and decreased to 4.74 U/L on tenth day.(2)The content of neutral detergent fiber in wheat straw were significantly decreased after fermentation with Pleurotus ostreatus 615 strain(P < 0.05),and decreased by 4.23% compared with the control group.The content of acid detergent fiber was significantly increased(P < 0.05),and increased by 4.19% compared with the control group. The content of cellulose was significantly higher than that of the control group(P < 0.01),and increased by 4.19% compared with the control group.The content of hemicellulose and lignin decreased significantly(P < 0.01),which was 19.87% and 43.75% lower than that of the control group. At the same time,the content of crude protein was significantly increased,which was 4.14% higher than that of the control group(P < 0.01).(3)The results of scanning electron microscopy showed that the crude fiber could be effectively degraded after the fermenting of the Pleurotus ostreatus 615 strain.The activity of laccase and manganese peroxidase in Pleurotus ostreatus 615 strain was higher on the fourth day and the fifth day.The contents of lignin and hemicellulose in wheat straw were significantly decreased and the content of crude protein was significantly increased after 30 days of solid fermentation by Pleurotus ostreatus 615 strain.
In this study,the fungus Pleurotus ostreatus 615 strain was fermented by solid-state fermentation of wheat straw,designed to study effects of Pleurotus ostreatus 615 strain on fiber degradation in wheat straw.Liquid culture of Pleurotus ostreatus 615 strain and the activity of laccase and manganese peroxidase in 1 ~ 10 days were determined.Then the wheat straw which treaded with 1.00% Ca O were fermented at 25 ℃ for 30 days. And the contents of neutral detergent fiber,acid detergent fiber,cellulose,hemicelluloses,lignin and crude protein of wheat straw were determined by solid fermentation of Pleurotus ostreatus 615 strain seed liquid.Last,the fermented wheat straw was scanned by the electron microscope to observe its structural changes.The results showed as follows:(1)Laccase activity increased from first day to fourth day and reached a maximum of 1428.00 U/L on fourth day,decreased from fifth day and creased to399.00 U/L on tenth day;manganese peroxidase activity increased from first day to fifth day and reached a maximum of 15.45 U/L on fifth day,decreased from sixth day and decreased to 4.74 U/L on tenth day.(2)The content of neutral detergent fiber in wheat straw were significantly decreased after fermentation with Pleurotus ostreatus 615 strain(P < 0.05),and decreased by 4.23% compared with the control group.The content of acid detergent fiber was significantly increased(P < 0.05),and increased by 4.19% compared with the control group. The content of cellulose was significantly higher than that of the control group(P < 0.01),and increased by 4.19% compared with the control group.The content of hemicellulose and lignin decreased significantly(P < 0.01),which was 19.87% and 43.75% lower than that of the control group. At the same time,the content of crude protein was significantly increased,which was 4.14% higher than that of the control group(P < 0.01).(3)The results of scanning electron microscopy showed that the crude fiber could be effectively degraded after the fermenting of the Pleurotus ostreatus 615 strain.The activity of laccase and manganese peroxidase in Pleurotus ostreatus 615 strain was higher on the fourth day and the fifth day.The contents of lignin and hemicellulose in wheat straw were significantly decreased and the content of crude protein was significantly increased after 30 days of solid fermentation by Pleurotus ostreatus 615 strain.
引文
[1]池玉杰,闫洪波.红平菇木质素降解酶系统漆酶、锰过氧化物酶及木质素过氧化物酶的检测[J].林业科学,2009,45(12):154~158.
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[17]张荣,奚永兰,叶小梅,等.棘孢木霉1285对麦秸的降解及厌氧发酵的影响[J].江苏农业学报,2015,31(2):328~333.
[18]朱顺国,邢壮,张微,等.玉米秸秆NDF与ADF含量变化规律的研究[J].中国奶牛,2001,1:24~26.
[19]Arora D S.Biodelignification of wheat straw by different fungal associations[J].Biodegradation,1995,6(1):57~60.
[20]Asiegbu F O,Paterson A,Smith J E.The effects of co-fungal cultures and supplementation with carbohydrate adjuncts on lignin biodegradation and substrate digestibility[J].World Journal of Microbiology&Biotechnology,1996,12(3):273~279.
[21]Hadar Y,Kerem Z,Gorodecki B,et al.Utilization of lignocellulosic waste by the edible mushroom,Pleurotus[J].Biodegradation,1992,3(2~3):189~205.
[22]Shi J,Chinn M S,Sharmashivappa R R.Microbial pretreatment of cotton stalks by solid state cultivation of Phanerochaete chrysosporium.[J].Bioresource Technology,2008,99(14):6556.
[23]Lv E,Xia W,Tang M,et al.Preparation of an efficient oil-spill adsorbent based on wheat straw[J].Bio Resources,2016,12(1):296~315.
[24]Wei Xiang W,Hao Hao L,Wei L,et al.Decomposition of Bacillus thuringiensis(Bt)transgenic criceresidues(straw and roots)in paddy fields[J].Journal of Soils and Sediments,2009,10(9):457~467.
[2]邓缘.浅谈秸秆木质素的生物降解[J].江西饲料,2005,5:29~31.
[3]范鹏珍,池玉杰,惠非琼,等.杏鲍菇和乳白耙齿菌锰过氧化物酶活性规律的研究[J].中国林副特产,2011,29(1):1~4.
[4]范文霞,蔡友华,刘学铭,等.毛云芝菌产漆酶液体培养条件的优化[J].食品与生物技术学报,2008,27(3):88~93.
[5]高振华,邸明伟.生物质材料及应用[M].北京:化学工业出版社,2008.
[6]哈丽代·热合木江,阿依古丽·达噶尔别克,扎依尔·艾山,等.粉碎或颗粒化棉花秸秆饲喂绵羊对增质量的影响[J].饲料研究,2014,13:52~53.
[7]侯进,李婷,李杰.白腐真菌降解玉米秸秆的研究[J].饲料博览,2010,1:4~7.
[8]侯进,李婷,吕文龙,等.应用白腐真菌发酵曲种发酵玉米秸秆的研究[J].中国奶牛,2010,7:25~27.
[9]李坚.生物质复合材料学[M].北京:科学出版社,2008.
[10]卢松.微生物处理玉米秸秆的腐解特征研究:[硕士学位论文][D].重庆:西南大学,2010.
[11]李旭东,荚荣,吕飒音,等.不同缓冲体系下p H值对裂褶菌F17产锰过氧化物酶的影响[J].安徽大学学报(自然科学版),2006,30(5):78~82.
[12]李燕荣.双孢蘑菇高效栽培及其对秸秆生物降解的研究:[硕士学位论文][D].湖南长沙:中南林业科技大学,2010.
[13]李野,梁军锋,张洪生,等.4株食用菌发酵小麦秸秆粗蛋白含量研究[J].饲料研究,2008,9:41~44.
[14]王炫清,苗嘉曦,陈兴,等.高效木质纤维素分解菌Aspergillus fumigatus Z5原位分解小麦秸秆的研究[J].南京农业大学学报,2017,40(4):671~680.
[15]王茂成.木质素降解真菌的筛选鉴定及相关酶活性研究:[硕士学位论文][D].重庆:西南大学,2013.
[16]魏敏.棉花秸杆对绵羊饲用价值的初步研究:[硕士学位论文][D].乌鲁木齐:新疆农业大学,2002.
[17]张荣,奚永兰,叶小梅,等.棘孢木霉1285对麦秸的降解及厌氧发酵的影响[J].江苏农业学报,2015,31(2):328~333.
[18]朱顺国,邢壮,张微,等.玉米秸秆NDF与ADF含量变化规律的研究[J].中国奶牛,2001,1:24~26.
[19]Arora D S.Biodelignification of wheat straw by different fungal associations[J].Biodegradation,1995,6(1):57~60.
[20]Asiegbu F O,Paterson A,Smith J E.The effects of co-fungal cultures and supplementation with carbohydrate adjuncts on lignin biodegradation and substrate digestibility[J].World Journal of Microbiology&Biotechnology,1996,12(3):273~279.
[21]Hadar Y,Kerem Z,Gorodecki B,et al.Utilization of lignocellulosic waste by the edible mushroom,Pleurotus[J].Biodegradation,1992,3(2~3):189~205.
[22]Shi J,Chinn M S,Sharmashivappa R R.Microbial pretreatment of cotton stalks by solid state cultivation of Phanerochaete chrysosporium.[J].Bioresource Technology,2008,99(14):6556.
[23]Lv E,Xia W,Tang M,et al.Preparation of an efficient oil-spill adsorbent based on wheat straw[J].Bio Resources,2016,12(1):296~315.
[24]Wei Xiang W,Hao Hao L,Wei L,et al.Decomposition of Bacillus thuringiensis(Bt)transgenic criceresidues(straw and roots)in paddy fields[J].Journal of Soils and Sediments,2009,10(9):457~467.