泡盛曲霉CAU33固体发酵产β-1,3-1,4-葡聚糖酶发酵条件优化
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摘要
利用单因素实验法优化了泡盛曲霉(Aspergillus awamori)CAU33利用农业废弃物固体发酵产β-1,3-1,4-葡聚糖酶的发酵条件。产酶的条件包括碳源种类、初始水分含量、氮源种类、初始p H、表面活性剂、培养温度和发酵时间。进一步运用响应面分析法优化了其中主要因素,得到最佳产酶条件为:啤酒糟为碳源、含水量为81.6%、吐温60添加量20g/L、大豆蛋白胨添加量25g/L、自然p H、35℃下培养6d。在优化后的发酵条件下,最大产酶水平达到40 832.9U/g。泡盛曲霉固体发酵产β-1,3-1,4-葡聚糖酶的酶活力高,工业化生产和应用潜力大。
The fermentation conditions for β-1,3-1,4-glucanase production from Aspergillus awamori CAU33 by solid state fermentation(SSF)with agricultural wastes as carbon source were optimized. Seven factors including carbon source, initial moisture content, nitrogen source, initial p H, surfactant source, temperature and fermentation time were optimized by single-factor experiment firstly, and the major factors amongst were then optimized by response surface method. The optimal fermentation conditions for β-1,3-1,4-glucanase production were as follows:beer sludge as carbon source, initial moisture content of 81.6%, Tween 60 of 20 g/L, soya peptone content of 25 g/L, natural p H, culture temperature of 35℃ and culture time of 6 d. Under the optimized conditions, the highest β-1,3-1,4-glucanase production of 40 832.9 U/g was achieved. The high yield may give the enzyme great potential in industrial applications.
The fermentation conditions for β-1,3-1,4-glucanase production from Aspergillus awamori CAU33 by solid state fermentation(SSF)with agricultural wastes as carbon source were optimized. Seven factors including carbon source, initial moisture content, nitrogen source, initial p H, surfactant source, temperature and fermentation time were optimized by single-factor experiment firstly, and the major factors amongst were then optimized by response surface method. The optimal fermentation conditions for β-1,3-1,4-glucanase production were as follows:beer sludge as carbon source, initial moisture content of 81.6%, Tween 60 of 20 g/L, soya peptone content of 25 g/L, natural p H, culture temperature of 35℃ and culture time of 6 d. Under the optimized conditions, the highest β-1,3-1,4-glucanase production of 40 832.9 U/g was achieved. The high yield may give the enzyme great potential in industrial applications.
引文
[1]PANDEY A,SOCCOL C R,MITCHELL D.New developments in solid state fermentation:I-bioprocesses and products[J].Process Biochemistry,2000,35(10):1153-1169.
[2]孙巍,刘学铭.酶的固体发酵生产研究进展[J].生物技术通报,20 08,2:64-67.
[3]MAKTOUF S,MOULIS C,KAMOUN A,et al.A laundry detergent compatible lichenase:statistical optimization for production under solid state fer mentation on cr ude millet[J].Industrial Crops and Products,2013,43:349-354.
[4]CHAARI F,KAMOUN A,BHIRI F,et al.Statistical optimization for the production of lichenase by a newly isolated Bacillus licheniformis UEB CF in solid state fermentation using pea pomace as a novel solid support[J].Industrial Crops and Products,2012,40:192-198.
[5]YANG S Q,XIONG H,YANG H Y,et al.High-level production ofβ-1,3-1,4-glucanase by Rhizomucor miehei u nder solid-st ate fermentation and its potential application in the brewing industry[J].Journal of Applied Microbiology,2015,118(1):84-91.
[6]LOWRY O H,ROSEBROUGH N J,FARR A L,et al.Protein measurement with folin phenol reagent[J].Jour nal of Biological Chemistry,1951,193(1):265-275.
[7]LAEMMLI U K.Cleavage of structural proteins during assembly of head of bacteriophage-T4[J].Nature,1970,227(5259):680-685.
[8]孙婕,郁惠蕾,李春秀,等.一株高产木聚糖酶真菌的筛选及酶学性质分析[J].生物加工过程,2013,11(1):35-40.
[9]BAKALOVA N G,PETROVA S D,KOLEV D N.Catalytically important amino acid residues in endoxylanases from Aspergillus Awamori[J].Biotechnology and Biotechnological Equipment,2009,23(23):639-642.
[10]TEIXEIRA R S S,SIQUEIRA F G,SOUZA M V D,et al.Purification and characterization studies of a thermostableβ-xylanase from Aspergillus awamori[J].Journal of Industrial Microbiology and Biotechnology,2010,37(10):1041-1051.
[11]马丽清.泡盛曲霉QH1β-甘露聚糖酶和β-甘露糖苷酶的研究[D].广州:华南农业大学,2007.
[12]MATSUBARA T,BEN A Y,ANINDYAWATI T,et al.Molecular clon i ng a nd deter m i nat ion of t he nucleot ide sequence of r aw starch digesting alpha-amylase from Aspergillus awamori KT-11[J].Jour nal of Biochemistr y and Molecular Biology,2004,37(4):429-438.
[2]孙巍,刘学铭.酶的固体发酵生产研究进展[J].生物技术通报,20 08,2:64-67.
[3]MAKTOUF S,MOULIS C,KAMOUN A,et al.A laundry detergent compatible lichenase:statistical optimization for production under solid state fer mentation on cr ude millet[J].Industrial Crops and Products,2013,43:349-354.
[4]CHAARI F,KAMOUN A,BHIRI F,et al.Statistical optimization for the production of lichenase by a newly isolated Bacillus licheniformis UEB CF in solid state fermentation using pea pomace as a novel solid support[J].Industrial Crops and Products,2012,40:192-198.
[5]YANG S Q,XIONG H,YANG H Y,et al.High-level production ofβ-1,3-1,4-glucanase by Rhizomucor miehei u nder solid-st ate fermentation and its potential application in the brewing industry[J].Journal of Applied Microbiology,2015,118(1):84-91.
[6]LOWRY O H,ROSEBROUGH N J,FARR A L,et al.Protein measurement with folin phenol reagent[J].Jour nal of Biological Chemistry,1951,193(1):265-275.
[7]LAEMMLI U K.Cleavage of structural proteins during assembly of head of bacteriophage-T4[J].Nature,1970,227(5259):680-685.
[8]孙婕,郁惠蕾,李春秀,等.一株高产木聚糖酶真菌的筛选及酶学性质分析[J].生物加工过程,2013,11(1):35-40.
[9]BAKALOVA N G,PETROVA S D,KOLEV D N.Catalytically important amino acid residues in endoxylanases from Aspergillus Awamori[J].Biotechnology and Biotechnological Equipment,2009,23(23):639-642.
[10]TEIXEIRA R S S,SIQUEIRA F G,SOUZA M V D,et al.Purification and characterization studies of a thermostableβ-xylanase from Aspergillus awamori[J].Journal of Industrial Microbiology and Biotechnology,2010,37(10):1041-1051.
[11]马丽清.泡盛曲霉QH1β-甘露聚糖酶和β-甘露糖苷酶的研究[D].广州:华南农业大学,2007.
[12]MATSUBARA T,BEN A Y,ANINDYAWATI T,et al.Molecular clon i ng a nd deter m i nat ion of t he nucleot ide sequence of r aw starch digesting alpha-amylase from Aspergillus awamori KT-11[J].Jour nal of Biochemistr y and Molecular Biology,2004,37(4):429-438.