公开(公告)号：US07235712B1

公开(公告)日：2007-06-26

申请号：US10203963

申请日：2000-02-15

申请人： Lian Hui Zhang ,  Xianzhen Li ,  Daohai Zhang

发明人： Lian Hui Zhang ,  Xianzhen Li ,  Daohai Zhang

IPC分类号： C12N15/31 ,  C12N15/82 ,  C12N15/61 ,  C12N15/62 ,  A01H5/00

CPC分类号： C12N9/90 ,  C12P19/12 ,  C12P19/24 ,  C12R1/22 ,  C12Y504/99011

摘要： Two strains of a novel bacterial species, Klebsiella singaporensis LX3 and LX21, are claimed. A nucleotide sequence (kis) encoding a novel form of isomaltulose synthase, KIS, is also claimed. Also claimed is a method for production of isomaltulose in a plant, which method comprises introducing into a cell of such plant a nucleic acid sequence which encodes an enzyme which converts sucrose into isomaltulose in a manner which allows said cell to express said nucleic acid sequence, as well as a functional cloning method of isolating nucleotide sequence encoding the KIS protein comprising the steps of (a) preparing a gene bank from a donor organism that contains a DNA sequence coding for an isomaltulose biosynthesis activity in a suitable host organism, (b) screening the clones of interest from the gene bank by their enhanced reducing sugar content, and (c) isolating the clones which contain a DNA coding for a protein with isomaltulose biosynthesis activity.

摘要翻译： 要求两种新型细菌物种，克莱伯氏菌（Klebsiella singaporensis）LX3和LX21。 还要求编码编码新型异麦芽酮糖合酶KIS的核苷酸序列（kis）。 还要求保护的是在植物中生产异麦芽酮糖的方法，该方法包括将这种核酸序列导入该植物的细胞中，所述核酸序列编码以允许所述细胞表达所述核酸序列的方式将蔗糖转化成异麦芽酮糖的酶， 以及分离编码KIS蛋白的核苷酸序列的功能性克隆方法，其包括以下步骤：（a）从供体生物体制备含有编码在合适宿主生物体中的异麦芽酮糖生物合成活性的DNA序列的基因库，（b） 通过其增加的还原糖含量筛选基因库中感兴趣的克隆，和（c）分离含有编码具有异麦芽酮糖生物合成活性的蛋白质的DNA的克隆。