台湾乳白蚁纤维素降解菌的分离和鉴定
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摘要
该研究从广西柳州本地的台湾乳白蚁(Coptotermes formosanus)体内筛选一株纤维素降解菌,通过形态观察、生理生化试验以及分子生物学对其进行鉴定,并采用滤纸片法及3,5-二硝基水杨酸(DNS)法测定该菌株的纤维素酶活力。结果表明,筛选获得一株可在以羧甲基纤维素钠为唯一碳源的培养基上生长的菌株,命名为BW,并鉴定其为约氏不动杆菌(Acinetobacter johnsonii);菌株BW的纤维素酶活力最高,为(93.73±0.22)U/m L。
A cellulose-degrading bacterium was screened from Coptotermes formosanus in Liuzhou, Guangxi, and identified by morphological observation, physiological and biochemical tests and molecular biology. The cellulase activity of the strain was measured by filter paper method and3,5-dinitrosalicylic acid(DNS) method. The results showed that the strain which could grew on the medium using carboxymethylcellulose sodium as the only carbon source was obtained, named as BW and identified as Acinetobacter johnsonii. The cellulase activity of strain BW was the highest and reached(93.73±0.22) U/ml.
A cellulose-degrading bacterium was screened from Coptotermes formosanus in Liuzhou, Guangxi, and identified by morphological observation, physiological and biochemical tests and molecular biology. The cellulase activity of the strain was measured by filter paper method and3,5-dinitrosalicylic acid(DNS) method. The results showed that the strain which could grew on the medium using carboxymethylcellulose sodium as the only carbon source was obtained, named as BW and identified as Acinetobacter johnsonii. The cellulase activity of strain BW was the highest and reached(93.73±0.22) U/ml.
引文
[1] DEMIRBAS A. Biofuels sources, biofuel policy, biofuel economy and global biofuel projections[J]. Energ Convers Manage, 2008, 49(8):2106-2116.
[2] LYND L R, LASER M S, BRANSBY D, et al. How biotech can transform biofuels[J]. Nat Biotechnol, 2008, 26(2):169-172.
[3]刘振东,李贵春,杨晓梅,等.我国农业废弃物资源化利用现状与发展趋势分析[J].安徽农业科学,2012,40(26):13068-13070.
[4] WYMAN C E. Biomass ethanol:technical progress, opportunities, and commercial challenges[J]. Annu Rev Energ Env, 1999, 24(1):189-226.
[5]孙建中,陈春润.昆虫与生物质能源利用:一个新的交叉学科前沿[J].昆虫知识,2010,47(6):1033-1042.
[6]刘冰,林景栋,宋水林,等.白蚁肠道可培养菌群及其功能初探[J].生物灾害科学,2012,35(2):172-176.
[7] CHRISTEL M, JULIEN B, CATHERINE B, et al. Identification and characterization of a new xylanase from Gram-positive bacteria isolated from termite gut(Reticulitermes santonensis)[J]. Protein Expres Purif,2012, 83(2):117-127.
[8] KUDO T. Termite-microbe symbiotic system and its efficient degradation of lignocellulose[J]. Biosci Biotech Bioch, 2009, 73(12):2561-2567.
[9]王巍.白蚁肠道木质素分解菌的分离鉴定及PY12 LiP基因的克隆与序列分析[D].长春:吉林农业大学,2013.
[10]章珍.黑翅土白蚁内源性纤维素酶基因在毕赤酵母的表达[D].杭州:浙江大学,2014.
[11]刘晓静.白蚁肠道元基因组来源β-葡萄糖苷酶bgl2、bgl15和bgl17的酶学性质研究[D].镇江:江苏科技大学,2015.
[12]黄小晖,付日辉,李德,等.白蚁肠道纤维素分解菌的筛选鉴定及产酶条件的研究[J].江西农业大学学报,2009,31(6):1140-1145.
[13]李丹红,徐荣,张坤迪,等.象白蚁肠道中一个纤维素降解菌群的分离和特性研究[J].生物资源,2017,39(4):272-278.
[14] POURRANMEZAN Z, GHEZELBASH G R, ROMANI B, et al. Screening and identification of newly isolated cellulose-degrading bacteria from the gut of xylophagous termite Microcerotermes diversus(Silvestri)[J].Microbiology, 2012, 81(6):736-742.
[15] TEIXEIRA L M, MERQUIOR V L C. The Family Moraxellaceae[M]//The Prokaryotes:Gammaproteobacteria. Rosenberg E, De Long E F, Lory S,(eds).. Berlin, Heidelberg:Springer, 2014:443-476.
[16] GHOSE T K. Measurement of cellulase activities[J]. Pure Appl Chem,1987, 59(2):257-268.
[17] PHILIPPE J M B, GRIMONT P. Taxonomy of the genus Acinetobacter with the recognition of Acinetobacter baumannii sp. nov. Acinetobacter haemolyticus sp. nov. Acinetobacter johnsonii sp. nov. and Acinetobacter junii sp. nov. and emended descriptions of Acinetobacter calcoaceticus and Acinetobacter lwofii[J]. Int J Syst Bacteriol, 1986, 36(2):228-240.
[18] XIE S, LIU J X, LI L, et al. Biodegradation of malathion by Acinetobacter johnsonii MA19 and optimization of cometabolism substrates[J]. J Environ Sci, 2009, 21(1):76-82.
[19] LEE M, WOO S G, TEN L N. Characterization of novel diesel-degrading strains Acinetobacter haemolyticus MJ01 and Acinetobacter johnsonii MJ4 isolated from oil-contaminated soil[J]. World J Microbiol Biotechn, 2012, 28(5):2057-2067.
[20] STRAGANZ G D, GLIEDER A, BRECHKER L, et al. Acetylacetonecleaving enzyme Dke1:a novel C-C-bond-cleaving enzyme from Acinetobacter johnsonii[J]. Biochem J, 2003, 369(3):573-581.
[21] HANNES H, THOMAS M, WALTER S. Diketone cleaving enzyme Dke1 production by Acinetobacter johnsonii-optimization of fermentation conditions[J]. J Biotechnol, 2004, 107(1):73-81.
[22] WANG H K, SHAO J, WEI Y J. et al. A novel low-temperature alkaline lipase from Acinetobacter johnsonii LP28 suitable for detergent formulation[J]. Food Technol Biotech, 2011, 49(1):96-102.
[23] HAI K W, JIE Z, XIAO J W. et al. Genome shuffling improves production of the low-temperature alkalophilic lipase by Acinetobacter johnsonii[J]. Biotechnol Lett, 2012, 34(1):145-151.
[24] SHIL R K, MOJUNMDER S, SADIDA F F. et al. Isolation and identification of cellulolytic bacteria from the gut of three phytophagus insect species[J]. Braz Arch Biol Techn, 2014, 6(57):927-932.
[25]李静,张瀚能,赵翀.等.高效纤维素降解菌分离筛选、复合菌系构建及秸秆降解效果分析[J].应用与环境生物学报,2016,22(4):689-696.
[26] EKPERIGIN M M. Preliminary studies of cellulase production by Acinetobacter anitratus and Branhamella sp.[J]. African J Biotechnol,2007, 6(1):28-33.
[2] LYND L R, LASER M S, BRANSBY D, et al. How biotech can transform biofuels[J]. Nat Biotechnol, 2008, 26(2):169-172.
[3]刘振东,李贵春,杨晓梅,等.我国农业废弃物资源化利用现状与发展趋势分析[J].安徽农业科学,2012,40(26):13068-13070.
[4] WYMAN C E. Biomass ethanol:technical progress, opportunities, and commercial challenges[J]. Annu Rev Energ Env, 1999, 24(1):189-226.
[5]孙建中,陈春润.昆虫与生物质能源利用:一个新的交叉学科前沿[J].昆虫知识,2010,47(6):1033-1042.
[6]刘冰,林景栋,宋水林,等.白蚁肠道可培养菌群及其功能初探[J].生物灾害科学,2012,35(2):172-176.
[7] CHRISTEL M, JULIEN B, CATHERINE B, et al. Identification and characterization of a new xylanase from Gram-positive bacteria isolated from termite gut(Reticulitermes santonensis)[J]. Protein Expres Purif,2012, 83(2):117-127.
[8] KUDO T. Termite-microbe symbiotic system and its efficient degradation of lignocellulose[J]. Biosci Biotech Bioch, 2009, 73(12):2561-2567.
[9]王巍.白蚁肠道木质素分解菌的分离鉴定及PY12 LiP基因的克隆与序列分析[D].长春:吉林农业大学,2013.
[10]章珍.黑翅土白蚁内源性纤维素酶基因在毕赤酵母的表达[D].杭州:浙江大学,2014.
[11]刘晓静.白蚁肠道元基因组来源β-葡萄糖苷酶bgl2、bgl15和bgl17的酶学性质研究[D].镇江:江苏科技大学,2015.
[12]黄小晖,付日辉,李德,等.白蚁肠道纤维素分解菌的筛选鉴定及产酶条件的研究[J].江西农业大学学报,2009,31(6):1140-1145.
[13]李丹红,徐荣,张坤迪,等.象白蚁肠道中一个纤维素降解菌群的分离和特性研究[J].生物资源,2017,39(4):272-278.
[14] POURRANMEZAN Z, GHEZELBASH G R, ROMANI B, et al. Screening and identification of newly isolated cellulose-degrading bacteria from the gut of xylophagous termite Microcerotermes diversus(Silvestri)[J].Microbiology, 2012, 81(6):736-742.
[15] TEIXEIRA L M, MERQUIOR V L C. The Family Moraxellaceae[M]//The Prokaryotes:Gammaproteobacteria. Rosenberg E, De Long E F, Lory S,(eds).. Berlin, Heidelberg:Springer, 2014:443-476.
[16] GHOSE T K. Measurement of cellulase activities[J]. Pure Appl Chem,1987, 59(2):257-268.
[17] PHILIPPE J M B, GRIMONT P. Taxonomy of the genus Acinetobacter with the recognition of Acinetobacter baumannii sp. nov. Acinetobacter haemolyticus sp. nov. Acinetobacter johnsonii sp. nov. and Acinetobacter junii sp. nov. and emended descriptions of Acinetobacter calcoaceticus and Acinetobacter lwofii[J]. Int J Syst Bacteriol, 1986, 36(2):228-240.
[18] XIE S, LIU J X, LI L, et al. Biodegradation of malathion by Acinetobacter johnsonii MA19 and optimization of cometabolism substrates[J]. J Environ Sci, 2009, 21(1):76-82.
[19] LEE M, WOO S G, TEN L N. Characterization of novel diesel-degrading strains Acinetobacter haemolyticus MJ01 and Acinetobacter johnsonii MJ4 isolated from oil-contaminated soil[J]. World J Microbiol Biotechn, 2012, 28(5):2057-2067.
[20] STRAGANZ G D, GLIEDER A, BRECHKER L, et al. Acetylacetonecleaving enzyme Dke1:a novel C-C-bond-cleaving enzyme from Acinetobacter johnsonii[J]. Biochem J, 2003, 369(3):573-581.
[21] HANNES H, THOMAS M, WALTER S. Diketone cleaving enzyme Dke1 production by Acinetobacter johnsonii-optimization of fermentation conditions[J]. J Biotechnol, 2004, 107(1):73-81.
[22] WANG H K, SHAO J, WEI Y J. et al. A novel low-temperature alkaline lipase from Acinetobacter johnsonii LP28 suitable for detergent formulation[J]. Food Technol Biotech, 2011, 49(1):96-102.
[23] HAI K W, JIE Z, XIAO J W. et al. Genome shuffling improves production of the low-temperature alkalophilic lipase by Acinetobacter johnsonii[J]. Biotechnol Lett, 2012, 34(1):145-151.
[24] SHIL R K, MOJUNMDER S, SADIDA F F. et al. Isolation and identification of cellulolytic bacteria from the gut of three phytophagus insect species[J]. Braz Arch Biol Techn, 2014, 6(57):927-932.
[25]李静,张瀚能,赵翀.等.高效纤维素降解菌分离筛选、复合菌系构建及秸秆降解效果分析[J].应用与环境生物学报,2016,22(4):689-696.
[26] EKPERIGIN M M. Preliminary studies of cellulase production by Acinetobacter anitratus and Branhamella sp.[J]. African J Biotechnol,2007, 6(1):28-33.