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1、农业资源与环境学报JournalofAgriculturalResourcesandEnvironment2016年5月·第33卷·第3期:262-267May2016·Vol.33·No.3:262-267产黄纤维单胞菌CR-14菌株诱变选育及发酵条件优化*杨继业,崔冠慧,习彦花,李亚冰,张根伟,张丽萍,程辉彩(河北省科学院生物研究所,河北石家庄050081)摘要:为提高产黄纤维单胞菌CR-14纤维素酶活力,对菌株进行亚硝酸、紫外线及复合诱变处理,筛选产纤维素酶活较高的突变株,并对其发酵条件进行优化。结
2、果表明,经亚硝酸和紫外线复合诱变得到一株产纤维素酶活较高的菌株Y-UA-18,其酶活力为10.57U,为原菌株产酶活力的1.67倍。通过正交试验得到菌株Y-UA-18产酶最佳培养基为:秸秆粉1.0%、复合氮源0.6%、KH2PO40.1%、MgSO40.1%、NaCl0.08%;最佳培养条件为:初始pH值7.0、培养温度30℃、发酵时间3d。通气量对菌株Y-UA-18产酶影响不明显,但在厌氧条件下发酵液酶活显著降低,0.1%TW-80对菌株Y-UA-18的产酶没有影响。关键词:产黄纤维单胞菌;纤维素酶;诱
3、变;发酵条件中图分类号:X172文献标志码:A文章编号:2095-6819(2016)03-0262-06doi:10.13254/j.jare.2015.0269引用格式:杨继业,崔冠慧,习彦花,等.产黄纤维单胞菌CR-14菌株诱变选育及发酵条件优化[J].农业资源与环境学报,2016,33(3):262-267.YANGJi-ye,CUIGuan-hui,XIYan-hua,etal.ScreeningandMutationBreedingoftheCelloulonmonasflavigenaCR-
4、14andOptimizationofItsFermentationConditions[J].JournalofAgriculturalResourcesandEnvironment,2016,33(3):262-267.ScreeningandMutationBreedingoftheCelloulonmonasflavigenaCR-14andOptimizationofItsFermentationConditionsYANGJi-ye,CUIGuan-hui,XIYan-hua,LIYa-bin
5、g,ZHANGGen-wei,ZHANGLi-ping,CHENGHui-cai*(InstituteofBiology,HebeiAcademyofSciences,Shijiazhuang050081,China)Abstract:TheCelloulonmonasflavigenaCR-14wasscreenedfromtherumenofcattle,whichhadtheabilityofproductingcelluloseandwasstoredinourlaboratory.Inorder
6、toimprovethecelluloseactivityoftheCR-14,itwastreatedwithnitrite,ultraviolet(UV)andcomplexmutagenesis.Initialselectionidentified6cellulase-producingstrains,asdemonstratedbytheirabilitytoturnCongoredplatesyellow.These6strainswerethenrescreenedbydeterminingt
7、hecellulaseactivityofbroth.Finally,throughnitriteandUVmutagenesis,Y-UA-18strainwiththehighcelluloseyieldandstablecharacteristics,whichenzymaticactivitywas10.57U,1.67timesofstartingstrainCR-14,wasscreenedout.Inordertoprovideatheoreticalreferencetotheindust
8、rialproductionofthestrainY-UA-18,weconductedthesinglefactortestandorthogonaltesttothefermentationmediumcomponentsandthefermentationconditions.TheresultsshowedthatthebestmediumofstrainY-UA-18enzymeproductionwasasfoll