响应面法优化灰色链霉菌产纤维素酶的工艺研究
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摘要
以灰色链霉菌为原料,在单因素试验的基础上,采用响应面法试验,优化灰色链霉菌产纤维素酶活性的发酵条件。结果表明,单因素试验灰色链霉菌产纤维素酶活性的最适发酵条件:碳源为CMC-Na,氮源为明胶,温度为28℃,pH为7.0,转速为130 r/min。响应面法试验优化灰色链霉菌产纤维素酶活性最佳发酵条件为:温度27.7℃,pH值6.9,转数130.3 r/min,在此优化条件下,灰色链霉菌产纤维素平均酶活性为6.103 U/mL(n=3),与模型的预测值(6.217 U/mL)比较接近,误差为1.83%,证明了该响应面模型具有可靠性。
This study took Streptomyces griseus as raw materials, and optimized the fermentation conditions of cellulase activity in the Streptomyces griseus by the response surface tests, on the basis of single factor test. The results showed that the optimum fermentation conditions of cellulase activity in the Streptomyces griseus by single factor test were determined as follows: carbon source was CMC-Na, the nitrogen source was gelatin, temperature was 28℃, pH was 7.0, and rotate speed was 130 r/min. The optimum fermentation conditions of cellulase activity in the Streptomyces griseus by response surface method were determined as follows: temperature was 27.7℃, pH was 6.9, and rotate speed was 130.3 r/min. Under the optimization condition, the average cellulase-producing activity of the Streptomyces griseus was 6.103 U/mL(n=3), which was close to the the predicted value of the model(6.217 U/mL), and the errors was 1.83%. It proved that the response surface model might be reliable.
This study took Streptomyces griseus as raw materials, and optimized the fermentation conditions of cellulase activity in the Streptomyces griseus by the response surface tests, on the basis of single factor test. The results showed that the optimum fermentation conditions of cellulase activity in the Streptomyces griseus by single factor test were determined as follows: carbon source was CMC-Na, the nitrogen source was gelatin, temperature was 28℃, pH was 7.0, and rotate speed was 130 r/min. The optimum fermentation conditions of cellulase activity in the Streptomyces griseus by response surface method were determined as follows: temperature was 27.7℃, pH was 6.9, and rotate speed was 130.3 r/min. Under the optimization condition, the average cellulase-producing activity of the Streptomyces griseus was 6.103 U/mL(n=3), which was close to the the predicted value of the model(6.217 U/mL), and the errors was 1.83%. It proved that the response surface model might be reliable.
引文
Bo D.P.,2000,Feed cellulase and microbes producing cellulase,Ningxia Nongxueyuan Xuebao(Journal of Ningxia Agricultural College),21(4):72-78(卜登攀,2000,饲用纤维素酶及产酶微生物,宁夏农学院学报,21(4):72-78)
Cao L.L.,Wang Y.,Tang S.K.,Zhang P.H.,Mao P.H.,Jin X.,Wang C.L.,and Lou K.,2009,Biodiversity and enzyme screening of actinomycetes from hami lake,Weishengwu Xuebao(Acta Microbiologica Sinica),49(3):287-293(曹兰兰,王芸,唐蜀昆,张评浒,毛培宏,金湘,王纯利,娄恺,2009,新疆哈密地区盐湖放线菌的多样性及其功能酶的筛选,微生物学报,49(3):287-293)
Chen W.X.,2007,Screening of cellulose-degrading actinomyces strain and studies on enzyme-producing conditions and characters of enzyme activity,Thesis for M.S.,Sichuan Normal University,Supervisor:Song H.,pp.9-19(陈文祥,2007,产纤维素酶放线菌的筛选及其产酶条件与酶学性质初探,硕士学位论文,四川师范大学,导师:宋浩,pp.9-19)
Deng H.,2013,Study on the diversity and cellulose degrading ability of actinomycetes involved in composting process,Thesis for M.S.,Zhejiang University,Supervisor:Wu W.X.,pp.9-19(邓辉,2013,堆肥过程放线菌多样性及其纤维素降解特性研究,硕士学位论文,浙江大学,导师:吴伟祥,pp.9-19)
Fang X.,Shen Y.,Qu Y.B.,Bao X.M.,and Qu Y.B.,2010,Status and prospect of lignocellulosic bioethanol production in China,Bioresour.Technol.,101:4814-4819
Guo C.,2010,Studies on screening for cellulose-degrading actionomyces and its functional identification,Thesis for M.S.,Fujian Agriculture and Forestry University,Supervisor:Lin W.X.,pp.34-48(郭昌,2010,高纤维素降解放线菌的筛选与功能鉴定,硕士学位论文,福建农林大学,导师:林文雄,pp.34-48)
Huang H.Q.,Huang M.R.,YE J.J.,Zhu J.,and Bao S.X.,2010,Screening and identification of two cellulase-producing actinomycetes,Weishengwuxue Zazhi(Journal of Microbiology),30(1):47-50(黄惠琴,黄美容,叶建军,朱军,鲍时翔,2010,2株产纤维素酶放线菌的筛选及分类鉴定,微生物学杂志,30(1):47-50)
Li S.W.,Kong K.,Feng J.L.,Li M.,and Tai X.S.,2010,Screening and identificatiaon of a cold-adapted cellulase-producing actinomycete and characterization of cellulase,Shengwujishu Tongbao(Biotechnolgy Bulletin),(7):216-220(李师翁,孔凯,冯佳丽,李梅,台喜生,2010,产低温纤维素酶放线菌的筛选、鉴定及酶学性质初步研究,生物技术通报,(7):216-220)
Li Z.L.,2008,Sereening of Alkaline cellulose-degrding actinomyces strain and studies on enzyme-producing conditions and characters of enzyme activity,Thesis for M.S.,Northwest University,Supervisor:Wang W.W.,pp.2-3(李忠玲,2008,产碱性纤维素酶放线菌的筛选及其产酶条件与酶学性质初步研究,硕士学位论文,西北大学,导师:王卫卫,pp.2-3)
Qin W.N.,Zhao K.,Guo L.Z.,and Lu W.D.,2016,Identification,process optimization of cellulase production halophilic actinomycete DN-A4,and characterization of the enzymatic properties of its cellulase,Qingdao Nongye Daxue Xuebao(Ziran Kexue Ban)(Journal of Qingdao Agricultural University(Natural Science)),33(1):27-34(秦雯娜,赵坤,郭立忠,卢伟东,2016,1株产纤维素酶嗜盐放线菌的鉴定、发酵优化及酶学性质的研究,青岛农业大学学报(自然科学版),33(1):27-34)
Shao N.N.,Xuan H.L.,Luo F.,and Dai X.Z.,2015,Isolation,identification and characterization of a cold-adapted cellulaseproducing psychrotrophic Streptomyces sp.,Shipin Gongye Keji(Science and Technology of Food Industry),36(24):159-168(邵娜娜,轩换玲,罗锋,代先祝,2015,一株产低温纤维素酶放线菌的分离与产酶研究,食品工业科技,36(24):159-168)
Tong Y.,2004,Studies on celluase-producing actionmy microbal communities in freshwater lakes,Thesis for M.S.,Chinese Academy of Sciences,Supervisor:Ke X.,pp.12-15(佟勇,2004,产纤维素酶放线菌及其酶学性质和淡水湖底泥微生物区系研究,硕士学位论文,中国科学院研究生院,导师:柯欣,pp.12-15)
Wei Y.H.,2009,Screening and identification of cellulase-producing strains and optimization of their fermentation conditions,Thesis for M.S.,Central China Normal University,Supervisor:Song H.,pp.10-25(魏艳红,2009,纤维素酶产生菌的分离鉴定及产酶条件优化,硕士学位论文,华中师范大学,导师:刘德立,pp.10-25)
Zang H.,Deng J.C.,and Xie Y.L.,2015,Screening and optimization of culture conditions of a high yield cellulase actinomycetes,Minjiang Xueyuan Xuebao(Journal of Minjiang U-niversity),(5):100-108(郑虹,邓加聪,谢颖灵,2015,一株高温纤维素酶放线菌的筛选及发酵条件优化,闽江学院学报,(5):100-108)
Zhang C.F.,Gu W.J.,Peng K.F.,Wang Y.Y.,Cao L.Q.,and Wang L.Q.,2007,Present situation of research on microbial cellulase,Shengwu Xinxixue(China Journal of Bioinformatics),5(1):34-36(张传富,顾文杰,彭科峰,吴永英,曹立群,王立群,2007,微生物纤维素酶的研究现状,生物信息学,5(1):34-36)
Zhang X.M.,Ma Y.N.,Wang S.,and Zhang L.P.,2011,Study on biodiversity and feature of enzyme excreting about endophytic actinomycetes isolated from Ophiopogon japonicus Ker-Gawl.,and Gynura japonica Juel.,Hebei Nongye Daxue Xuebao(Journal of Agricultural University),34(3):50-53,55,(张秀敏,马友楠,王顺,张利平,2011,麦冬和菊三七药用植物内生放线菌的多样性及产酶特性的研究,河北农业大学学报,34(3):50-53,55)
Cao L.L.,Wang Y.,Tang S.K.,Zhang P.H.,Mao P.H.,Jin X.,Wang C.L.,and Lou K.,2009,Biodiversity and enzyme screening of actinomycetes from hami lake,Weishengwu Xuebao(Acta Microbiologica Sinica),49(3):287-293(曹兰兰,王芸,唐蜀昆,张评浒,毛培宏,金湘,王纯利,娄恺,2009,新疆哈密地区盐湖放线菌的多样性及其功能酶的筛选,微生物学报,49(3):287-293)
Chen W.X.,2007,Screening of cellulose-degrading actinomyces strain and studies on enzyme-producing conditions and characters of enzyme activity,Thesis for M.S.,Sichuan Normal University,Supervisor:Song H.,pp.9-19(陈文祥,2007,产纤维素酶放线菌的筛选及其产酶条件与酶学性质初探,硕士学位论文,四川师范大学,导师:宋浩,pp.9-19)
Deng H.,2013,Study on the diversity and cellulose degrading ability of actinomycetes involved in composting process,Thesis for M.S.,Zhejiang University,Supervisor:Wu W.X.,pp.9-19(邓辉,2013,堆肥过程放线菌多样性及其纤维素降解特性研究,硕士学位论文,浙江大学,导师:吴伟祥,pp.9-19)
Fang X.,Shen Y.,Qu Y.B.,Bao X.M.,and Qu Y.B.,2010,Status and prospect of lignocellulosic bioethanol production in China,Bioresour.Technol.,101:4814-4819
Guo C.,2010,Studies on screening for cellulose-degrading actionomyces and its functional identification,Thesis for M.S.,Fujian Agriculture and Forestry University,Supervisor:Lin W.X.,pp.34-48(郭昌,2010,高纤维素降解放线菌的筛选与功能鉴定,硕士学位论文,福建农林大学,导师:林文雄,pp.34-48)
Huang H.Q.,Huang M.R.,YE J.J.,Zhu J.,and Bao S.X.,2010,Screening and identification of two cellulase-producing actinomycetes,Weishengwuxue Zazhi(Journal of Microbiology),30(1):47-50(黄惠琴,黄美容,叶建军,朱军,鲍时翔,2010,2株产纤维素酶放线菌的筛选及分类鉴定,微生物学杂志,30(1):47-50)
Li S.W.,Kong K.,Feng J.L.,Li M.,and Tai X.S.,2010,Screening and identificatiaon of a cold-adapted cellulase-producing actinomycete and characterization of cellulase,Shengwujishu Tongbao(Biotechnolgy Bulletin),(7):216-220(李师翁,孔凯,冯佳丽,李梅,台喜生,2010,产低温纤维素酶放线菌的筛选、鉴定及酶学性质初步研究,生物技术通报,(7):216-220)
Li Z.L.,2008,Sereening of Alkaline cellulose-degrding actinomyces strain and studies on enzyme-producing conditions and characters of enzyme activity,Thesis for M.S.,Northwest University,Supervisor:Wang W.W.,pp.2-3(李忠玲,2008,产碱性纤维素酶放线菌的筛选及其产酶条件与酶学性质初步研究,硕士学位论文,西北大学,导师:王卫卫,pp.2-3)
Qin W.N.,Zhao K.,Guo L.Z.,and Lu W.D.,2016,Identification,process optimization of cellulase production halophilic actinomycete DN-A4,and characterization of the enzymatic properties of its cellulase,Qingdao Nongye Daxue Xuebao(Ziran Kexue Ban)(Journal of Qingdao Agricultural University(Natural Science)),33(1):27-34(秦雯娜,赵坤,郭立忠,卢伟东,2016,1株产纤维素酶嗜盐放线菌的鉴定、发酵优化及酶学性质的研究,青岛农业大学学报(自然科学版),33(1):27-34)
Shao N.N.,Xuan H.L.,Luo F.,and Dai X.Z.,2015,Isolation,identification and characterization of a cold-adapted cellulaseproducing psychrotrophic Streptomyces sp.,Shipin Gongye Keji(Science and Technology of Food Industry),36(24):159-168(邵娜娜,轩换玲,罗锋,代先祝,2015,一株产低温纤维素酶放线菌的分离与产酶研究,食品工业科技,36(24):159-168)
Tong Y.,2004,Studies on celluase-producing actionmy microbal communities in freshwater lakes,Thesis for M.S.,Chinese Academy of Sciences,Supervisor:Ke X.,pp.12-15(佟勇,2004,产纤维素酶放线菌及其酶学性质和淡水湖底泥微生物区系研究,硕士学位论文,中国科学院研究生院,导师:柯欣,pp.12-15)
Wei Y.H.,2009,Screening and identification of cellulase-producing strains and optimization of their fermentation conditions,Thesis for M.S.,Central China Normal University,Supervisor:Song H.,pp.10-25(魏艳红,2009,纤维素酶产生菌的分离鉴定及产酶条件优化,硕士学位论文,华中师范大学,导师:刘德立,pp.10-25)
Zang H.,Deng J.C.,and Xie Y.L.,2015,Screening and optimization of culture conditions of a high yield cellulase actinomycetes,Minjiang Xueyuan Xuebao(Journal of Minjiang U-niversity),(5):100-108(郑虹,邓加聪,谢颖灵,2015,一株高温纤维素酶放线菌的筛选及发酵条件优化,闽江学院学报,(5):100-108)
Zhang C.F.,Gu W.J.,Peng K.F.,Wang Y.Y.,Cao L.Q.,and Wang L.Q.,2007,Present situation of research on microbial cellulase,Shengwu Xinxixue(China Journal of Bioinformatics),5(1):34-36(张传富,顾文杰,彭科峰,吴永英,曹立群,王立群,2007,微生物纤维素酶的研究现状,生物信息学,5(1):34-36)
Zhang X.M.,Ma Y.N.,Wang S.,and Zhang L.P.,2011,Study on biodiversity and feature of enzyme excreting about endophytic actinomycetes isolated from Ophiopogon japonicus Ker-Gawl.,and Gynura japonica Juel.,Hebei Nongye Daxue Xuebao(Journal of Agricultural University),34(3):50-53,55,(张秀敏,马友楠,王顺,张利平,2011,麦冬和菊三七药用植物内生放线菌的多样性及产酶特性的研究,河北农业大学学报,34(3):50-53,55)
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