一株低温秸秆纤维素降解菌的分离、鉴定及降解特性
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摘要
本研究在18℃条件下,利用赫奇逊滤纸培养基分离纯化菌株,根据透明圈和滤纸降解情况筛选菌株,最终通过酶活分析和秸秆降解率确定目的菌株,通过形态学观察与ITS基因序列分析对目的菌株进行初步鉴定,分离筛选高效低温秸秆纤维素降解菌并研究其产酶特性。结果表明,从小兴安岭山区土壤中分离到1株在低温下具有较强纤维素降解能力的真菌菌株C1,15 d内对秸秆的降解能力达55.6%,滤纸酶活和CMC酶活分别为18.4 U/mL和54.3 U/mL,初步鉴定菌株C1为青霉菌属(Penicillium sp.),具有进一步研究开发的潜力。
This study aimed to isolate and screen straw-cellulose degradation strain adapted for low-tempera-ture. At 18℃, we isolated and purified many strains by Hutchinson medium, further screened them according to the transparent circles and filter paper degradation, and then determined the target one through enzyme activity analysis and straw-cellulose degradation rate. The selected strain was preliminarily identified by its morphological character-istics, and then preformed internal transcribed spacers(ITS) gene sequence analysis. Finally, we successfully ob-tained a fungus from Xiaoxing'an Mountains collections and named it as C1, which was highly efficiency for cellu-lose degradation at low temperature, with straw-cellulose degradation rate of 55.6% within 15 d, and filter paper and CMC enzyme activities being 18.4 and 54.3 U/mL, respectively. Strain C1 was identified as a kind of Penicillium sp., which has the potential of further research.
This study aimed to isolate and screen straw-cellulose degradation strain adapted for low-tempera-ture. At 18℃, we isolated and purified many strains by Hutchinson medium, further screened them according to the transparent circles and filter paper degradation, and then determined the target one through enzyme activity analysis and straw-cellulose degradation rate. The selected strain was preliminarily identified by its morphological character-istics, and then preformed internal transcribed spacers(ITS) gene sequence analysis. Finally, we successfully ob-tained a fungus from Xiaoxing'an Mountains collections and named it as C1, which was highly efficiency for cellu-lose degradation at low temperature, with straw-cellulose degradation rate of 55.6% within 15 d, and filter paper and CMC enzyme activities being 18.4 and 54.3 U/mL, respectively. Strain C1 was identified as a kind of Penicillium sp., which has the potential of further research.
引文
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[18]尚晓瑛,程旭艳,霍培书,等.1株堆肥耐低温纤维素降解菌的筛选、鉴定及生长特性的初步研究[J].华中农业大学学报,2012,31(5):558-562.Shang X Y,Cheng X Y,Huo P S,et al.Screening,identification and growth of a cold-adapted cellulose-decomposing bacterium for composting[J].Journal of Huazhong Agricultural University,2012,31(5):558-562.(in Chinese)
[19]宋文芳,胡国全,徐彦胜.一株低温厌氧纤维素降解细菌的分离、鉴定及其酶学性质的研究[J].西南农业学报,2011,24(4):1317-1322.Song W F,Hu G Q,Xu Y S.Isolation,identification and enzymic characteristics of cold-adapted,cellulytic,anaerobic bacterium[J].Southwest China Journal of Agricultural Sciences,2011,24(4):1317-1322.(in Chinese)
[20]孟建宇,李蘅,樊兆阳,等.低温纤维素降解菌的分离与鉴定[J].应用与环境生物学报,2014,20(1):152-156.Meng J Y,Li H,Fan Z Y,et al.Isolation and identification of cellulose-degrading bacteria under low temperature[J].Chin J.Appl Environ Biol,2014,20(1):152-156.(in Chinese)
[2]杨茜,李维尊,鞠美庭,等.微生物降解木质纤维素类生物质固废的研究进展[J].微生物学通报,2015,42(8):1569-1583.Yang Q,Li W Z,Ju M T,et al.Advances in microbial degradation of lignocelluloses biomass solid waste-a review[J].Microbiology China,2015,42(8):1569-1583.(in Chinese)
[3]冯健玲,姚晓华,韦秉兴,等.稻草秸秆纤维素分解菌的分离筛选[J].基因组学与应用生物学,2009,28(3):477-480.Feng J L,Yao X h,Wei B X,et al.Isolation and screening of straw cellulose-decomposing microbes[J].Genomics and applied biology,2009,28(3):477-480.(in Chinese)
[4]赵旭,王文丽,李娟,等.低温秸秆降解微生物菌剂的研究进展[J].生物技术通报,2014,11(6):55-61.Zhao X,Wang W L,Li J,et al,Advanced in microbiological agent of straw degradation under low temperature[J].Biotechnology Bulletin,2014,11(6):55-61.(in Chinese)
[5]张壮志.高效纤维素分解菌的筛选、鉴定及其生长条件和产酶活性的研究[D].山东农业大学,2009.
[6]钱林,郑巧利,付瑾,等.一株高效纤维素降解菌株的分离鉴定及其酶学性质[J].微生物学通报,2010,37(4):524-528.Qian L,Zheng Q L,Fu J,et al.Isolation and identification of a cellulose degrading bacterium and its cellulase characterization[J].Microbiology China,2010,37(4):524-528.(in Chinese)
[7]穆春雷,武晓森,李术娜,等.低温产纤维素酶菌株的筛选、鉴定及纤维素酶学性质[J].微生物学通报,2013,40(7):1193-1201.Mu C L,Wu X S,Li S N,et al.Screening and identification of a cold-adapted cellulase-producing strains and characterization of cellulase[J].Microbiology China,2013,40(7):1193-1201.(in Chinese)
[8]张建峰.东北地区秸秆降解工程菌的选育及速腐菌剂的研制[D].吉林农业大学,2012.
[9]王洪媛,范丙全.三株高效秸秆纤维素降解真菌的筛选及其降解效果[J].微生物学报,2010,50(7):870-875.Wang H Y,Fan B Q.Screening of three straw-cellulose degrading microorganism[J].Acta Microbiologica Sinica,2010,50(7):870-875.(in Chinese)
[10]付秋玥.低温纤维素降解菌的筛选及其混合发酵效果研究[D].东北农业大学,2013.
[11]Coward-Kelly G,Aiello-Mazzari C,Kim S,et al.Suggested improvements to the standard filter paper assay used to measure cellulose activity[J].Biotechnology and Bioengineering,2003,82(6):745-749.
[12]Mandels M,Sternbrg D.Recent advances in cellulose technology[J].Journal of Fermentation Technology,1976,54(4):267-286.
[13]黄春凯,左小明,王红蕾,等.一株产纤维素酶菌株的分离、鉴定及产酶特性[J].微生物学通报,2015,42(4):646-653.Huang C K,Zuo X M,Wang H L,et al.Isolation,identification and characterization of a cellulase-producing strain[J].Microbiology China,2015,42(4):646-653.(in Chinese)
[14]张瑞萍.纤维素酶的滤纸酶活和CMC酶活的测定[J].印染助剂,2002,19(5):51-53.Zhang R P.Determination of filter papere enzyme activity and CMC enzyme activity of cellulase[J].Textile Auxiliaries,2002,19(5):51-53.(in Chinese)
[15]魏景超.真菌鉴定手册[M].上海:上海科学技术出版社,1979.
[16]孔华忠.中国真菌志[M].北京:科学出版社,2007.
[17]许玉林,郑月霞,叶冰莹,等.一株纤维素降解真菌的筛选及鉴定[J].微生物学通报,2013,40(2):220-227.Xu Y L,Zheng Y X,Ye B Y,et al,Isolation and identification of a cellulose degrading fungi[J].Microbiology China,2013,40(2):220-227.(in Chinese)
[18]尚晓瑛,程旭艳,霍培书,等.1株堆肥耐低温纤维素降解菌的筛选、鉴定及生长特性的初步研究[J].华中农业大学学报,2012,31(5):558-562.Shang X Y,Cheng X Y,Huo P S,et al.Screening,identification and growth of a cold-adapted cellulose-decomposing bacterium for composting[J].Journal of Huazhong Agricultural University,2012,31(5):558-562.(in Chinese)
[19]宋文芳,胡国全,徐彦胜.一株低温厌氧纤维素降解细菌的分离、鉴定及其酶学性质的研究[J].西南农业学报,2011,24(4):1317-1322.Song W F,Hu G Q,Xu Y S.Isolation,identification and enzymic characteristics of cold-adapted,cellulytic,anaerobic bacterium[J].Southwest China Journal of Agricultural Sciences,2011,24(4):1317-1322.(in Chinese)
[20]孟建宇,李蘅,樊兆阳,等.低温纤维素降解菌的分离与鉴定[J].应用与环境生物学报,2014,20(1):152-156.Meng J Y,Li H,Fan Z Y,et al.Isolation and identification of cellulose-degrading bacteria under low temperature[J].Chin J.Appl Environ Biol,2014,20(1):152-156.(in Chinese)