一株纤维素酶产生菌的筛选与产酶特性研究
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摘要
纤维素降解菌在纤维素物质的降解和资源化利用中具有重要作用。利用传统微生物学方法,从腐烂秸秆中筛选出一株高产纤维素酶的真菌QL06,通过形态学特征与ITSDNA序列同源性分析,该株菌为烟曲霉(Aspergillusfumigatus),以单因素试验与正交试验对产酶条件进行优化。结果表明,QL06培养84 h所产CMC酶活和生物量均最大,以麸皮+葡萄糖为碳源、酵母膏为氮源时有利于QL06产酶;在接种量体积分数为8%、麸皮+葡萄糖质量分数2%、酵母膏质量分数1.5%、初始pH值为4.0,培养温度40℃时,CMC酶活力达5.41 U/mL,具有进一步开发利用的价值。
Cellulose-degrading bacteria play an important role in the degradation and resource utilization of cellulosic materials. In this paper, a high-yield cellulase-producing fungus QL06 was screened from decayed straw by traditional microbiological method. The morphological characteristics and ITS DNA sequence homology analysis showed that the strain was Aspergillus fumigatus with single factor. The enzyme production conditions were optimized by experiments and orthogonal experiments. The results showed that the activity and biomass of CMC produced by QL06 cultured for 84 h were the highest. When Bran + glucose was used as carbon source and yeast extract was used as nitrogen source, it was beneficial to QL06 production enzyme; inoculation amount volume fraction was 8%, bran + glucose mass fraction 2%, yeast extract mass fraction 1.5%, initial pH 4.0,CMC enzyme activity reached 5.41 U/mL at a culture temperature of 40 ℃, which has further development and utilization value.
Cellulose-degrading bacteria play an important role in the degradation and resource utilization of cellulosic materials. In this paper, a high-yield cellulase-producing fungus QL06 was screened from decayed straw by traditional microbiological method. The morphological characteristics and ITS DNA sequence homology analysis showed that the strain was Aspergillus fumigatus with single factor. The enzyme production conditions were optimized by experiments and orthogonal experiments. The results showed that the activity and biomass of CMC produced by QL06 cultured for 84 h were the highest. When Bran + glucose was used as carbon source and yeast extract was used as nitrogen source, it was beneficial to QL06 production enzyme; inoculation amount volume fraction was 8%, bran + glucose mass fraction 2%, yeast extract mass fraction 1.5%, initial pH 4.0,CMC enzyme activity reached 5.41 U/mL at a culture temperature of 40 ℃, which has further development and utilization value.
引文
[1]LIU D Y,ZHANG R F,YANG X M,et al.Thermostable cellulase production of Aspergillus fumigatus Z5 under solid-state fermentation and its application in degradation of agricultural wastes[J].International biodeterioration&biodegradation,2011,65(5):717-725.
[2]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.
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[4]胡亚林,冼亮,刘果,等.两株纤维素降解真菌的分离鉴定及其产纤维素酶酶学性质的初步研究[J].南方农业学报,2008,39(4):425-428.
[5]张瑾,翟付群,刘学擎.徐州及周边地区秸秆综合处理方式的调查[J].环境科技,2009,22(S1):115-117.
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[7]KE Z,PEI Z,WANG D.Organic solvent pretreatment of lignocellulosic biomass for biofuels and biochemicals:Areview[J].Bioresour technol,2016,199:21-33.
[8]JANGA K K,HAGG M B,MOE S T.Influence of acid concentration,temperature,and time on decrystallization in twostage concentrated sulfuric acid hydrolysis of pinewood and aspenwood:astatistical approach[J].Bioresources,2012,7(1):391-411.
[9]王冠颖.酶在饲料中的应用研究进展[J].饲料博览,2016(1):34-37.
[10]杨捷,严芬,叶秀云,等.一株产高温纤维素酶曲霉菌的发酵条件及培养基优化[J].福州大学学报(自然科学版),2013,41(1):115-120.
[11]周俊强,邱忠平,韩云平,等.纤维素降解菌的筛选及其产酶特性[J].环境工程学报,2010,4(3):705-708.
[12]AHEMD S,BASHIR A,SALEEM H,et al.Production and purification of cellulose-degrading enzymes from a filamentou[J].Brazilian journal of microbiology,2009,41(3):1 411-1 419.
[13]欧阳蒲月,郭成栓,黄智璇,等.一株纤维素降解菌的分离,鉴定及产酶条件初步研究[J].现代生物医学进展,2009,9(18):3 447-3 450.
[14]金伟,陈文静,姚斌,等.一株产纤维素酶的耐热烟曲霉筛选及产酶条件研究[J].中国酿造,2012,31(6):61-64.
[15]GOMES A F S,SANTOS B S L,FRANCISCON E G,et al.Substract and temperature effect on xylanase production by aspergillus fumigatus using low cost agricultural wastes[J].Bioscience j,2016,32(4):915-921.
[16]ANG S K,SHAZA E M,ADIBAH Y,et al.Production of cellulases and xylanase by Aspergillus fumigatus,SK1 using untreated oil palm trunk through solid state fermentation[J].Process biochemistry,2013,48(9):1 293-1 302.
[17]CHERIAN E,KALPANA R,JAYALAKSHMI E.Production of cellulase by Aspergillus fumigatus JCF using agro-based cellulosic waste and its usage in bioethanol synthesis[J].International journal of energy technology&policy,2015,11(3):294-302.
[18]CHERIAN E,KUMAR M D,BASKAR G.Characterization of newly isolated strain Aspergillus fumigatus JCF from spoiled jackfruit and optimization of cellulase for bioethanol production using fruit waste[J].Journal of environmental biology,2015,36(5):1 229-1 237.
[19]赵淮阳,章梅丽,阿依妮尕尔·艾尔肯,等.江苏东海县小麦秸秆和猪粪堆肥及原位还田试验研究[J].环境科技,2015,28(3):1-6.
[20]SANTOS B S L D,GOMES A F S,FRANCISCON E G,et al.Thermotolerant and mesophylic fungi from sugarcane bagasse and theirprospection for biomass-degrading enzyme production[J].Brazilian journal of microbiology,2015,46(3):903-910.
[21]亢宗静,袁楠,王蓥燕,等.若尔盖高原湿地的真菌群落结构及低温纤维素降解真菌特征[J].土壤通报,2017,48(4):830-836.
[22]陈红漫,孙玉辉,阚国仕,等.产耐热纤维素酶菌株的分子鉴定及产酶条件优化[J].食品与发酵工业,2011,37(7):28-33.
[23]DIAS L M,DOS B S,ALBUQUERQUE C,et al.Biomass sorghum as a novel substrate in solid state fermentation for the production of hemicellulases and cellulases by Aspergillus niger and A.fumigatus[J].Journal of applied microbiology,2018,124(3):708-718.
[24]JUWON A D,EMMANUEL O F.Experimental investigations on the effects of Carbon and nitrogen sources on concomitant amylase and polygalacturonase production by trichoderma viride BITRS‐1001 in Submerged Fermentation[J].Biotechnol research international,2012,2012:1-8.
[2]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.
[3]FANG X,SHEN Y J,BAO X,et al.Status and prospect of lignocellulosic bioethanol production in China[J].Bioresource technology,2010,101(13):4 814-4 819.
[4]胡亚林,冼亮,刘果,等.两株纤维素降解真菌的分离鉴定及其产纤维素酶酶学性质的初步研究[J].南方农业学报,2008,39(4):425-428.
[5]张瑾,翟付群,刘学擎.徐州及周边地区秸秆综合处理方式的调查[J].环境科技,2009,22(S1):115-117.
[6]SYEIN J H,GUSTAV V,BJORGE W,et al.Novel enzymes for the degradation of cellulose[J].Biotechnology for biofuels,2012,5(1):1-12.
[7]KE Z,PEI Z,WANG D.Organic solvent pretreatment of lignocellulosic biomass for biofuels and biochemicals:Areview[J].Bioresour technol,2016,199:21-33.
[8]JANGA K K,HAGG M B,MOE S T.Influence of acid concentration,temperature,and time on decrystallization in twostage concentrated sulfuric acid hydrolysis of pinewood and aspenwood:astatistical approach[J].Bioresources,2012,7(1):391-411.
[9]王冠颖.酶在饲料中的应用研究进展[J].饲料博览,2016(1):34-37.
[10]杨捷,严芬,叶秀云,等.一株产高温纤维素酶曲霉菌的发酵条件及培养基优化[J].福州大学学报(自然科学版),2013,41(1):115-120.
[11]周俊强,邱忠平,韩云平,等.纤维素降解菌的筛选及其产酶特性[J].环境工程学报,2010,4(3):705-708.
[12]AHEMD S,BASHIR A,SALEEM H,et al.Production and purification of cellulose-degrading enzymes from a filamentou[J].Brazilian journal of microbiology,2009,41(3):1 411-1 419.
[13]欧阳蒲月,郭成栓,黄智璇,等.一株纤维素降解菌的分离,鉴定及产酶条件初步研究[J].现代生物医学进展,2009,9(18):3 447-3 450.
[14]金伟,陈文静,姚斌,等.一株产纤维素酶的耐热烟曲霉筛选及产酶条件研究[J].中国酿造,2012,31(6):61-64.
[15]GOMES A F S,SANTOS B S L,FRANCISCON E G,et al.Substract and temperature effect on xylanase production by aspergillus fumigatus using low cost agricultural wastes[J].Bioscience j,2016,32(4):915-921.
[16]ANG S K,SHAZA E M,ADIBAH Y,et al.Production of cellulases and xylanase by Aspergillus fumigatus,SK1 using untreated oil palm trunk through solid state fermentation[J].Process biochemistry,2013,48(9):1 293-1 302.
[17]CHERIAN E,KALPANA R,JAYALAKSHMI E.Production of cellulase by Aspergillus fumigatus JCF using agro-based cellulosic waste and its usage in bioethanol synthesis[J].International journal of energy technology&policy,2015,11(3):294-302.
[18]CHERIAN E,KUMAR M D,BASKAR G.Characterization of newly isolated strain Aspergillus fumigatus JCF from spoiled jackfruit and optimization of cellulase for bioethanol production using fruit waste[J].Journal of environmental biology,2015,36(5):1 229-1 237.
[19]赵淮阳,章梅丽,阿依妮尕尔·艾尔肯,等.江苏东海县小麦秸秆和猪粪堆肥及原位还田试验研究[J].环境科技,2015,28(3):1-6.
[20]SANTOS B S L D,GOMES A F S,FRANCISCON E G,et al.Thermotolerant and mesophylic fungi from sugarcane bagasse and theirprospection for biomass-degrading enzyme production[J].Brazilian journal of microbiology,2015,46(3):903-910.
[21]亢宗静,袁楠,王蓥燕,等.若尔盖高原湿地的真菌群落结构及低温纤维素降解真菌特征[J].土壤通报,2017,48(4):830-836.
[22]陈红漫,孙玉辉,阚国仕,等.产耐热纤维素酶菌株的分子鉴定及产酶条件优化[J].食品与发酵工业,2011,37(7):28-33.
[23]DIAS L M,DOS B S,ALBUQUERQUE C,et al.Biomass sorghum as a novel substrate in solid state fermentation for the production of hemicellulases and cellulases by Aspergillus niger and A.fumigatus[J].Journal of applied microbiology,2018,124(3):708-718.
[24]JUWON A D,EMMANUEL O F.Experimental investigations on the effects of Carbon and nitrogen sources on concomitant amylase and polygalacturonase production by trichoderma viride BITRS‐1001 in Submerged Fermentation[J].Biotechnol research international,2012,2012:1-8.
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