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    • 4. 发明申请
    • KETO GROUP INTRODUCING ENZYME, DNA CODING FOR THE SAME, AND PROCESS FOR PRODUCING KETOCAROTENOID
    • KETO集团介绍ENZYME,DNA编码及其生产方法KETOCAROTENOID
    • WO1996006172A1
    • 1996-02-29
    • PCT/JP1995001640
    • 1995-08-18
    • KIRIN BEER KABUSHIKI KAISHAKAJIWARA, SusumuMISAWA, NorihikoKONDO, Keiji
    • KIRIN BEER KABUSHIKI KAISHA
    • C12N15/53
    • C12N9/0004C12P7/26C12P23/00
    • A polypeptide having the enzymatic activity of converting the 4-methylene group of a beta -ionone compound into a keto group; a DNA containing the base sequence coding for the above polypeptide; another DNA which hybridizes with the above DNA and contains the base sequence coding for the above polypeptide; still another DNA which has been inserted into plasmid pHP51 and contains the base sequence coding for the above polypeptide; a recombinant vector containing the above DNA(s); a microbe having the above DNA(s) introduced thereinto; and a process for producing a ketocarotenoid which comprises culturing the above microbe in a medium and separating the formed ketocarotenoid from the product of culture. The introduction of the above DNAs as foreign genes into microbes, such as E. coli, followed by expression thereof makes it possible to impart to the microbes the capability of biosynthesis of astaxanthin, 4-ketozeaxanthin, canthaxanthin, echinenone and other ketocarotenoids. The use of such microbes makes it possible to mass-produce ketocarotenoids at reduced cost and labor.
    • 具有将β-戊酮化合物的4-亚甲基转化为酮基的酶活性的多肽; 含有编码上述多肽的碱基序列的DNA; 与上述DNA杂交并含有编码上述多肽的碱基序列的另一种DNA; 仍然是已经插入质粒pHP51并且含有编码上述多肽的碱基序列的另一个DNA; 含有上述DNA的重组载体; 将上述DNA引入其中的微生物; 以及生产酮类胡萝卜素的方法,其包括在培养基中培养上述微生物,并将所形成的酮类胡萝卜素与培养产物分离。 将上述DNA作为外源基因导入微生物,如大肠杆菌,随后进行表达,可以使微生物赋予虾青素,4-酮基黄豆素,角黄素,海参酮等酮类胡萝卜素的生物合成能力。 使用这样的微生物使得可以以降低的成本和劳动量大量生产酮类胡萝卜素。
    • 5. 发明申请
    • YEAST VECTORS AND PROCESS FOR PRODUCING PROTEINS WITH THE USE OF THE SAME
    • YEAST VECTORS和使用该蛋白的蛋白质生产过程
    • WO1998007873A1
    • 1998-02-26
    • PCT/JP1997002924
    • 1997-08-22
    • KIRIN BEER KABUSHIKI KAISHAKONDO, KeijiMIURA, Yutaka
    • KIRIN BEER KABUSHIKI KAISHA
    • C12N15/68
    • C12N9/2411C07K14/43C12N9/2408C12N15/815C12N15/905C12Y302/01001
    • Vectors which are to be integrated onto yeast chromosomes by multi-copying; and vectors which are to be integrated onto yeast chromosomes by multi-copying and show improved stability of expression units on the chromosomes. These vectors are ones each containing a marker gene for selecting a transformant, a shortened promoter sequence linked to the marker gene to thereby serve as an operator, and a sequence homologous with the chromosomal DNA of Candida utilis, optionally together with a heterogenous gene or a gene originating in C. utilis. It is linearized by cleaving with restriction enzymes within the homologous DNA sequence or at both ends of the same and, thus, enables the integration of the heterogenous gene or the gene originating in C. utilis onto the chromosomal DNA of C. utilis via homologous recombination.
    • 通过多重复制整合到酵母染色体上的载体; 以及通过多次复制整合到酵母染色体上的载体,并且显示改善染色体上表达单元的稳定性。 这些载体是含有用于选择转化体的标记基因,与标记基因连接的缩短的启动子序列,从而用作操纵子的序列,以及与产朊假丝酵母的染色体DNA同源的序列,任选与异源基因或 源自产朊酵母的基因。 通过用同源DNA序列内的限制性内切酶或同源DNA两端的限制性内切酶线性化,从而通过同源重组使异源基因或源自产朊酵母的基因与产朊假单胞菌的染色体DNA整合 。