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1、福建農(nóng)業(yè)學(xué)報29(8):757~767,2014文章編號:1008-0384(2014)08-757-11FujianJournalofAgriculturalSciences劉程程,劉波,藍(lán)江林,等.產(chǎn)木聚糖酶芽胞桿菌的篩選及產(chǎn)酶條件優(yōu)化[J].福建農(nóng)業(yè)學(xué)報,2014,29(8):757-767.LIUC-C,LIUB,LANJ-L,etal.ScreeningofXylanase-producingBacillussp.andOptimizationofFermentationProcess[J].FujianJournalofA
2、griculturalSciences,2014,29(8):757-767.產(chǎn)木聚糖酶芽胞桿菌的篩選及產(chǎn)酶條件優(yōu)化劉程程1,2,劉波1,2*,藍(lán)江林2,王凱3(1.福建農(nóng)林大學(xué)生命科學(xué)學(xué)院,福建福州350003;2.福建省農(nóng)業(yè)科學(xué)院農(nóng)業(yè)生物資源研究所,福建福州350003;3.福州大學(xué)生物工程學(xué)院,福建福州350002)摘要:利用木聚糖酶選擇培養(yǎng)基,從120株標(biāo)準(zhǔn)芽胞桿菌中篩選出37株能夠形成明顯透明圈的芽胞桿菌。經(jīng)產(chǎn)酶發(fā)酵液培養(yǎng)基復(fù)篩,得到1株木聚糖酶活性較高的菌株FJAT-14260(沙福芽胞桿菌Bacillussafensis
3、)。采用單因素實(shí)驗(yàn)設(shè)計和正交實(shí)驗(yàn)設(shè)計進(jìn)行優(yōu)化,結(jié)果表明,在培養(yǎng)基優(yōu)化中對產(chǎn)酶影響最大的因素是底物濃度;在發(fā)酵條件優(yōu)化中對產(chǎn)酶影響最大的因素是培養(yǎng)溫度。優(yōu)化得到FJAT-14260的最佳產(chǎn)酶發(fā)酵條件為:碳源(酵母粉)7g·L-1,氮源(蛋白胨)9g·L-1,底物(木聚糖)濃度10g·L-1,培養(yǎng)溫度30℃,初始pH值8,裝液量30mL/250mL。在此條件下FJAT-14260在發(fā)酵32h時酶活力達(dá)到113585.78U·mL-1,比優(yōu)化前提高了4.62倍。關(guān)鍵詞:芽胞桿菌;木聚糖酶;發(fā)酵條件中圖分類號:Q939文獻(xiàn)標(biāo)識碼:AScree
4、ningofXylanase-producingBacillussp.andOptimizationofFermentationProcess1,2,LIUBo1,2*,LANJiang-lin2,WANGKai3LIUCheng-cheng(1.CollegeofLifeScience,FujianAgricultureandForestryUniversity,Fuzhou,Fujian350002,China;2.AgriculturalBio-ResourcesResearchInstitute,FujianAcademyof
5、AgriculturalSciences,Fuzhou,Fujian350003,China;3.CollegeofBiologicalScienceandTechnology,FuzhouUniversity,Fuzhou,Fujian350002,China)Abstract:SelectivemediumforxylanasewasusedinscreeningthexylanaseproducingBacillusstrains.Thirty-sevenofthe120Bacillusstrainstestedwerefoun
6、dtoformtransparentcirclesontheplates.Afterasecondaryscreeningwiththeenzymefermentationmedium,B.safensisFJAT-14260wasdeterminedtobethemostdesirablestrainforthexylanaseproduction.Asinglefactorandanorthogonalexperimentwereusedtooptimizethefermentationconditions.Theresultss
7、howedthesubstrateconcentrationtobethemostimportantfactorrelatingtothemedium,andtemperature,themostcriticalconditionforthefermentationprocess.Inall,thebestmediumfor-1,nitrogensource(peptone)at9g·theenzymeproductionconsistedofcarbonsource(yeastextract)at7g·L-1,andsubstrat
8、e(xylan)at10g·L-1L.And,theoptimalfermentationtemperaturewas30℃,initialpHat8.0,andliquidvolumeat30mL/250mL.Unde