∗doi:10.3969/j.issn.2095-1736.2023.05.069固定化表面展示锰过氧化物酶对甲基橙的吸附特性王艳君1,2,黄佳瑶1,陈少煌1,赵鹏飞1(1.福建技术师范学院食品与生物工程学院,福清350300;2.食品软塑包装技术福建省高校工程研究中心,福清350300)摘要将酵母细胞表面展示载体pYD1-MnP经LiAC法转化至酿酒酵母(Saccharomycescerevisiae)EBY100,经SDS-PAGE及免疫荧光检测表明,MnP蛋白已成功锚定于酵母细胞表面,体外诱导转化子产锰过氧化物酶活性。为偶氮染料废水的降解提供优良的微生物资源。经2,6-二甲氧基苯酚(DMP)法测定酶活并通过选取不同培养时间、培养温度、pH、碳源、氮源、碳氮源的浓度等因素对酶活影响进行测定,并以海藻酸钠将游离酶进行固定化。锚定成功的转化子PM-2体外诱导产锰过氧化物酶的最适培养时间为60h、诱导产酶最适温度为40℃;最适pH值为4.5,最适碳源为半乳糖,最适氮源为YNB,碳源浓度为0.02g/mL,氮源浓度为0.0072g/mL;经海藻酸钠固定化后对甲基橙模拟废水进行吸附性能研究,吸附动力学表明,吸附过程符合准二级方程;与Freundlich等温吸附方程拟合效果更佳。锚定成功的转化子PM-2具有产锰过氧化物酶活性的能力,可在偶氮染料的生物降解过程中发挥作用。关键词表面展示;锰过氧化物酶;免疫荧光;固定化;甲基橙中图分类号Q554+.6文献标识码A文章编号2095-1736(2023)05-0069-08ImmobilizedsurfacesdemonstratetheadsorptionpropertiesofmanganeseperoxidaseonmethylorangeWANGYanjun12HUANGJiayao1CHENShaohuang1ZHAOPengfei11.SchoolofFoodandBioengineeringFujianPolytechnicNormalUniversityFuqing350300China2.FujianUniversitiesandCollegesEngineeringResearchCenterofSoftPlasticPackagingTechnologyforFoodFuqing350300ChinaAbstractTheyeastcellsurfacedisplayvectorpyd1MnPwastransformedintoEBY100byLiACmethod.TheMnPproteinwassuc-cessfullyanchoredonthesurfaceofyeastcellsbySDS-PAGEandimmunofluorescenceandthetransformantwasinducedtoproducemanganeseperoxidaseactivityinvitro.Itcanprovideexcellentmicrobialresourcesforthedegradationofazodyewastewater.Theen-zymeactivitywasdeterminedby26-dimethoxyphenolDMPmethodandtheeffectsofdifferentculturetimetemperaturepHcar-bonsourcenitrogensourceandconcentrationofcarbonandnitrogensourceonenzymeactivityweredetermined.Theenzymewasim-mobilizedwithsodiumalginate....
