发酵对鲜湿米粉品质影响研究进展

米粉发酵后可获得更好的口感和品质。了解发酵对米粉理化性质的影响,不仅可以为传统米粉发酵工艺的改良提供理论依据,还可促进现代生物技术的介入和应用。本文综述了发酵米粉的微生物及工艺,以及发酵对大米的淀粉、蛋白质、脂肪和最终米粉产品品质影响方面的研究进展,对于选育出优良发酵菌种,改善米粉发酵品质进而可以进行大规模的工业化生产起到重要的作用。

Expression and Purification of E.coli NrfA Protein and Preparation of Polyclonal Antibody against NrfA

[Objective] This study aimed to clone the E. coil NrfA gene and construct the pET-28a （＋）-NrfA prokaryotic expression vector for preparation of polyclonal anti- body against E. coil NrfA. [Method] E. coil NrfA gene was cloned from the E. coli genome DNA by PCR and inserted into the vector pET-28a（＋） to construct prokary- otic expression vector pET-28a （＋）-NrfA. E. coil NrfA protein was expressed by IPTG induction and purified. Polyclonal antibody against NrfA protein was prepared by im- munizing rabbit with routine method. The specificity and titer of polyclonal antibody was confirmed by ELISA and Western Blotting. [Result] The constructed prokaryotic expression vector pET-28a（＋）-NrfA was induced by IPTG, the recombinant NrfA pro- tein could be expressed effectively. The titer of rabbit anti-NrfA polyclonal antibody obtained by immunization and purification was about 1：204 900. Western Blotting anal- ysis indicated that the obtained polyclonal antibody against E. coil NrfA protein had high titer and high specificity. [Conclusion] E. coil NrfA gene was cloned and the prokaryotic expression vector pET-28a （＋）-NrfA was constructed successfully, poly- clonal antibody with high titer and high specificity was prepared, which laid the foun- dation for the study of NrfA in different strains of bacteria.

大肠杆菌NrfA蛋白表达、纯化及多克隆抗体的制备

[目的]克隆大肠杆菌NrfA基因,构建pET-28a(+)-NrfA表达载体,制备相应的多克隆抗体并对其进行鉴定。[方法]以大肠杆菌基因组DNA为模板,PCR扩增得到NrfA基因编码区,构建pET-28a(+)-NrfA表达载体;经IPTG诱导表达并纯化重组蛋白;再免疫新西兰雄兔,制备多克隆抗体;用ELISA方法检测抗体的效价,Western Blotting检测抗体的特异性。[结果]构建的表达载体pET-28a(+)-NrfA在大肠杆菌中诱导后可高效表达NrfA蛋白;免疫获得的多克隆抗体用ELISA检测,其效价为1∶204 900;经Western Blotting分析,抗体的特异性较好。[结论]成功克隆大肠杆菌的NrfA基因,并构建了其表达载体,制备的NrfA多克隆抗体具有较高的效价和良好的特异性,为研究细菌有关NrfA奠定了基础。

11α-羟基坎利酮生物转化中副产物的影响因素

基于质谱数据推测11α-羟基坎利酮生物转化中的副产物为双羟坎利酮。考察了投料浓度、转化时间、装液量、转速、投料时间和后期转化温度对副产物生成的影响。得到副产物生成量较小的工艺条件为:坎利酮浓度6 g/L,转化时间60 h,转速160 r/min,装液量100 ml/500 ml,后期转化温度28℃,投料时间20 h,11α-羟基坎利酮的转化率为94%。