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以刺槐豆内生菌Paenibacillus sp.CH-3为出发菌株,采用紫外-硫酸二乙酯-亚硝酸盐对其进行复合诱变,并对菌株的发酵条件进行了优化。结果表明:经过紫外线-硫酸二乙酯-亚硝酸盐复合诱变,获得一株高产β-甘露聚糖酶的突变菌株Paenibacillus sp.CH3-05,其酶活力为160.2U/mL,较出发菌株(42U/mL)提高了281.4%。其最佳发酵培养基为:酵母膏0.25%,魔芋粉3%,磷酸氢二铵0.25%,氯化钠0.1%,硫酸镁0.3%,磷酸氢二钾0.2%,CaCl22 mmol/L。最佳发酵条件为:初始pH 6.5,接种量2%,培养温度35℃,转速200 r/min,培养时间78 h,在该条件下测得酶活为233.5 U/mL,较出发菌株提高456%。
Abstract：The mutagenic effects of ultraviolet-diethyl sulfate-nitrite for compound mutagenesis on the original strain Paenibacillus sp. CH-3(42 U/mL) screened from locust bean were studied. Furthermore, the fermentation conditions of strain were optimized. Results of compound mutagenesis showed that a mutant strain(160.2 U/mL)with a high-yield β-mannanase production, named Paenibacillus sp. CH3-05, was obtained after the ultraviolet-diethyl sulfate-nitrite compound mutagenesis. The activity of mutant strain increased 281.4% compared with that of the original strain Paenibacillus sp. CH-3. The optimal medium for β-mannanase production were determined as follows: Yeast extract 0.25%, konjaku flour 3%,(NH4)2HPO40.25%, NaCl 0.1%, MgSO40.3%, K2HPO40.2%,CaCl22 mmol/L. The best fermentation condition was as following: Initial pH value 6.5, inoculum size 2%, culture temperature 35℃, rotate speed 200 r/min, culture time 78 h. Under these circumstances, the highest enzyme activity was 233.5 U/mL. The activity of mutant strain increased 456% in comparison with that original strain.
Chau han P.S.,Puri N.,Sharma P.,and Gupta N.,2012,Mannanases:Microbial sources,production,properties and potential biotechnological applications,Applied Microbiology and Biotechnology,93(5):1817-1830
Han K.F.,20 13,Mutation breeding and purification of β-mannanase from an endophyic Paenibacillus sp.CH-3,Thesis for M.S.,Henan Normal University,Supervisor:Zhang J.X.,pp.9-31(韩科芳,2013,β-甘露聚糖酶内生类芽孢杆菌诱变选育及酶的纯化研究,硕士学位论文,河南师范大学,导师:张建新,pp.9-31)
Liang L.,Qiu Y.L.,and Xu J.T.,2008,Breeding of L-histidine producing bacterium by composite mutation with UV and NaNO2,Weishengwuxue Zazhi(Journal of Microbiology),28(2):27-29(梁亮,邱雁临,许进涛,2008,紫外线与亚硝酸钠复合诱变选育L-组氨酸产生菌,微生物学杂志,28(2):27-29)
Lin Y.Q.,and Hong W.,2012,Research and future application of plant endophytic fungi,Fujian Linye Keji(Journal of Fujian Forestry Science and Technology),39(3):186-193(林燕青,洪伟,2012,植物内生真菌研究及应用前景,福建林业科技,39(3):186-193)
Pan Y.,Zhang Y.B.,Guo Z.M.,Wang S.Y.,Xing H.M.,Lin F.,Teng L.R.,and Hou A.L.,2012,Study on screening and mutation of β-glucosidase-producing strain,Shipin Yanjiu YuKaifa(Food Research and Development),33(5):149-152(潘艳,张益波,郭志敏,王诗雨,邢慧萌,林风,滕利荣,侯阿澧,2012,β-葡萄糖苷酶高产菌株筛选、诱变及其发酵的研究,食品研究与开发,33(5):149-152)
Qin S.,Xing K.,Jiang J.H.,Xu L.H.,and Li W.J.,2011,Biodiversity,bioactive natural products and biotechnological potential of plant-associated endophytic actinobacteria,Applied Microbiology and Biotechnology,89(3):457-473
Shi Q.Q.,and Wu S.G.,eds.,2009,Genentic improvement of industrial microorganisms(3rd edition),Science Press,Beijing,China,pp.37-50(施巧琴,吴松刚,主编,2009,工业微生物育种学(第三版),科学出版社,中国,北京,pp.37-50)
Srivastava P.K.,and Kapoor M.,2013,Extracellular endo-mannanase from Bacillus sp.CFR1601:Economical production using response surface methodology and downstream processing using aqueous two phase system,Food and Bioproducts Processing,91(4):672-681
Wang L.H.,2012,Research advances on diversity of plant endophytes and their secondary metabolism active substances,Huaxue Yu Shengwu Gongcheng(Chemistry&Bioengineering),29(10):1-3(王莉衡,2012,植物内生菌及其次生代谢活性物质多样性的研究进展,化学与生物工程,29(10):1-3)
Wang Y.,and Dai C.C.,2011,Endophytes:A potential resource for biosynthesis,biotransformation,and biodegradation,Annals of Microbiology,61(2):207-215
Wang Y.,Tian C.H.,Zhu L.,and Zhu H.H.,2011,Compound inducement of Bacillus cereus strain producing isomerous mannanase,Jiangxi Nongye Xuebao(Acta Agriculturae Jiangxi),23(6):113-115(王永,田春华,竹磊,朱海华,2011,产异甘露聚糖酶菌株的复合诱变,江西农业学报,23(6):113-115)
Xu L.L.,Shan Q.H.,and Guo B.,2013,Endophytes research progress and application prospect,Anhui Nongye Kexue(Journal of Anhui Agricultural Sciences),41(13):5641-5643,5709(徐玲玲,单庆红,郭斌,2013,植物内生菌研究进展及应用展望,安徽农业科学,41(13):5641-5643,5709)
Xu Y.J.,2011,Research process on pesources diversity of plantendo phytes,Guangdong Nongye Kexue(Guangdong Agricultural Sciences),38(24):149-152(徐亚军,2011,植物内生菌资源多样性研究进展,广东农业科学,38(24):149-152)
Zhang J.X.,Zhang D.,Wang H.L.,Hu W.B.,Han K.F.,Zhu Y.K.,Zhao J.,and Nie G.X.,2012,Mutation breeding of a β-mannanase-producing endophytic bacteria and its enzymatic characterization,Xiandai Shipin Keji(Modern Food Scienceand Technology),28(10):1331-1335,1410(张建新,张吨,王海磊,胡文波,韩科芳,朱岩昆,赵杰,聂国兴,2012,产β-mannanase内生菌的诱变育种和酶学特性研究,现代食品科技,28(10):1331-1335,1410)
Zhang S.F.,Wu M.C.,Sheng J.P.,Zhu J.,and Li J.F.,2008,Mutation breeding of Aspergillus niger LW-1 for producinghigh-yield acidicβ-mannanase,Nongye Shengwu Jishu Xuebao(Journal of Agricultural Biotechnology),16(2):346-350(张树飞,邬敏辰,盛金萍,朱劼,李剑芳,2008,酸性β-甘露聚糖酶高产菌株的诱变育种,农业生物技术学报,16(2):346-350)
基本信息:
DOI：10.13417/j.gab.033.000815
中图分类号:Q93
引用信息:
[1]张建新,赵杰,陈泽田,等.产β-甘露聚糖酶内生菌的诱变育种及产酶条件优化[J].基因组学与应用生物学,2014,33(04):815-821.DOI:10.13417/j.gab.033.000815.
基金信息:
河南省重点科技攻关(132102310293,142102210453,112102110118);; 河南省教育厅科学技术研究重点项目(14B180023);; 河南师范大学国家级科研项目培育基金(2013PL10)共同资助
2014-08-28
2014-08-28