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1、h响应面法优化(R)-α-羟基苯乙酸培养基#陈秉梅,许小平**(福州大学化学化工学院,福建,福州,350116)510摘要:目的:采用响应面分析法对菌株Bacillussp.ULi-11发酵产(R)-α-羟基苯乙酸培养基进行优化。方法:采用Plackett-Burman试验设计筛选出影响发酵产率的三个主要因素,采用最陡爬坡路径逼近最大响应区域,再用Box-Behnken试验设计及响应面分析法进行回归分析。结果:最佳培养基浓度分别为:酪蛋白胨12.81g/L,淀粉12.74g/L,KH2PO42.97g/L,K2HPO41.2g/L,M
2、gSO40.15g/L,CaCl20.125g/L,MnSO40.015g/L,ZnSO40.01g/L。在优化条件下,(R)-α-羟基苯乙酸产率达69.78%,比优化前提高了37.6%。结论:响应面分析法用于寻求(R)-α-羟基苯乙酸发酵的最佳培养基组分浓度是可行的。关键词:(R)-α-羟基苯乙酸;Plackett-Burman;响应面分析法;培养基优化;Bacillussp.中图分类号:TQ015.915Optimizationof(R)-α-HydroxyphenylaceticAcidFermentationMediumbyUsi
3、ngResponseSurfaceMethodologyCHENBingmei,XUXiaoping2025303540(CollegeofChemistryandChemicalEngineeing,FuzhouUniversity,Fujian350116,China)Abstract:Purpose:ResponseSurfaceMethodologywasappliedtooptimizetheculturecomponentsfor(R)-α-hydroxyphenylaceticacidbyBacillussp.ULi-11
4、.Methods:Bytheinitialscreeningmethod,Plackett-Burmandesignwasusedtoevaluatetheinfluenceoftotaleightrelatedfactors.Itshowedthatthreefactorsplayingtheimportantrolesinthemedium.Pathofsteepestascentwasundertakentoapproachtheoptimalregionofthefermentationconditionssubsequently
5、.Finally,theconcentrationsofthosethreemainfactorswerefurtheroptimizedbyusingBox-BehnkenandResponseSurfaceAnalysis.Results:Bysolvingthequadraticregressionmodelequation,theoptimalconcentrationsofthevariablesweredeterminedas:starch12.81g/L,caseinpeptone12.74g/L,KH2PO42.97g/L
6、,K2HPO41.2g/L,MgSO40.15g/L,CaCl20.125g/L,MnSO40.015g/L,ZnSO40.01g/L.Theyieldof(R)-α-hydroxyphenylaceticacid(69.78%)waspromotedby37.6%attheoptimalcultureconditions,comparedtothatobtainedfrominitialfermentationconditions.Conclusion:Itisavailabletofindtheoptimalmediumfor(R)-
7、α-hydroxyphenylaceticacidbyresponsesurfacemethodology.Keywords:(R)-α-HydroxyphenylaceticAcid,Plackett-Burman,ResponseSurfaceMethodology,CultureMediumOptimization,Bacillussp.0引言(R)-α-羟基苯乙酸((R)-HPA)及其衍生物是一类重要的手性合成砌块,是制备诸如半合成[1]对映体(R)-HPA主要由化学合成和拆分法制备,但生物转化法具有高效、温和、污染少、选择性高等
8、优点,是手性药物研究的热点。本课题组筛选到一株芽孢杆菌,HPA产率为50.7%,对映体过量值e.e.>99%。为了进一步提高产率,本文采用Plackett-Burman设计确定培养基的主效因素