重组布氏乳杆菌漆酶表达的研究  

Study on Expression of Recombinant Laccase Lactobacillus Buchneri

作　　者：刘晓庆[1,2] 那日[2] 郭九峰[2] 薛丹[1,2] Liu Xiaoqing Na Ri Guo Jiufeng Xue Dan(School of Physical Science and Technology, Inner Mongolia University, Hohhot, 010021 Key Laboratory of Biotechnology, Inner Mongolia Au tonomous University Ion Beam, Hohhot, 010021)

机构地区：[1]内蒙古大学,物理科学与技术学院,呼和浩特010021 [2]内蒙古大学自治区离子束生物工程重点实验室,呼和浩特010021

出　　处：《基因组学与应用生物学》2016年第9期2404-2410,共7页Genomics and Applied Biology

基　　金：国家自然科学基金(51267014)资助

摘　　要：本研究将杏鲍菇漆酶基因接入食品级宿主细胞乳酸杆菌中,成功构建重组工程乳酸菌。为筛选出单个转化菌株,将重组工程乳酸菌液涂布到含有红霉素的固体MRS培养基中,挑取白色菌落直接进行PCR,筛选阳性重组子。并提取筛选到的阳性重组子质粒,进一步进行PCR验证。将验证过确实存在漆酶基因的阳性重组子涂布到含有愈创木酚的固体培养基中进行培养,菌落颜色变成红棕色表明具有分泌漆酶的能力,挑取颜色变化较深的菌落,测定其酶活。成功筛选到具有产漆酶能力的重组工程乳酸菌,对其适宜温度、酸碱度进行了研究。具有分泌漆酶能力的乳酸菌若能作为青贮饲料的添加剂,将大大提高青贮饲料的利用率及品质。In the research, we put the mushroom laccase gene into the prokaryotic cell lactic acid bacteria, and constructed the recombinant engineering lactic acid bacteria successfully. In order to screen out the single transformed plant. We culture the recombinant Lactobacillus liquid into solid MRS culture medium containing erythromycin, then pick the white colony direcltly to PCR for screening positive restructuring. Extract and select the positive recombinant plasmid, then further verified by the PCR. Then cuture the positive recombinant with has been tested containing laccase gene in the solid medium containing guaiacol. If the color of colony change into red brown suggests that it has the ability to secrete laccase, picking colonies which color is deeper and measureing it, senzyme activity. The research has been get the lactic acid bacteria restructuring project witch has the ability to produce laccse successful, and the optimum temperature and pH were studied. If the lactic acid bacteria restructuring project witch has the ability to produce laccse can be as silage additives, will improve the efficiency and the quality of silage greatly.

关 键 词：漆酶 乳酸菌 阳性重组子 酶活性 

分 类 号：Q936[生物学—微生物学] Q78