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    Process for producing lactonase and utilization thereof
    32.
    发明申请
    Process for producing lactonase and utilization thereof 审中-公开
    生产内酯酶的方法及其利用

    公开(公告)号：US20070134775A1

    公开(公告)日：2007-06-14

    申请号：US10547424

    申请日：2004-03-03

    申请人： Sakayu Shimizu ,  Michihiko Kataoka ,  Keiji Sakamoto

    发明人： Sakayu Shimizu ,  Michihiko Kataoka ,  Keiji Sakamoto

    IPC分类号： C12P17/06 ,  C12N9/16 ,  C12N1/18

    CPC分类号： C12P41/003 ,  C12N9/18 ,  C12P17/04

    摘要： It has been required to economically produce a large amount of optically active γ-lactone derivatives (for example, pantolactone), which are useful as intermediates in synthesizing useful substances such as pharmaceutical drugs. To achieve this object, it is advantageous to employ an enzymatic technique of asymmetric hydrolysis with a hydrolyzing enzyme lactonase. However, it remains troublesome to prepare the enzyme or utilize a microorganism capable of producing the enzyme. Also, it is difficult to obtain a sufficient and stable enzymatic activity in the case of using recombination techniques. In producing lactonase, which asymmetrically hydrolyzes a γ-lactone derivative such as racemic pantolactone selectively, by a recombination technique, both mature lactonase DNA and signal peptide region DNA are transferred into a host. Thus, a stable lactonase activity comparable to naturally-occurring one can be achieved and a transformant sustaining a high enzyme activity in a stable manner can be acquired, thereby allowing the efficient and industrially advantageous asymmetric synthesis of γ-lactone derivatives.

    摘要翻译： 需要经济地生产大量的用作合成有用物质如药物的中间体的光学活性γ-内酯衍生物（例如泛酸内酯）。 为了达到这个目的，使用水解酶内酯酶的不对称水解酶技术是有利的。 然而，制备酶或利用能够产生酶的微生物仍然是麻烦的。 此外，在使用重组技术的情况下，难以获得足够稳定的酶活性。 在制备内含子选择性地水解γ-内酯衍生物如外消旋泛酸内酯的内切酶中，通过重组技术，将成熟内含酶DNA和信号肽区DNA两者转移到宿主中。 因此，可以获得与天然存在的活性相当的稳定的内含酶活性，并且可以获得以稳定的方式维持高酶活性的转化体，从而允许有效和工业上有利的γ-内酯衍生物的不对称合成。

    D-pantolactone hydrolase and gene encoding the same
    35.
    发明授权
    D-pantolactone hydrolase and gene encoding the same 失效
    D-泛酸内酯水解酶和编码基因的基因

    公开(公告)号：US06794171B2

    公开(公告)日：2004-09-21

    申请号：US10133372

    申请日：2002-04-29

    申请人： Keiji Sakamoto ,  Hideaki Yamada ,  Sakayu Shimizu ,  Michihiko Kobayashi

    发明人： Keiji Sakamoto ,  Hideaki Yamada ,  Sakayu Shimizu ,  Michihiko Kobayashi

    IPC分类号： C12N914

    CPC分类号： C12N9/18 ,  C12P17/04 ,  C12P41/001 ,  C12P41/005

    摘要： A novel enzyme which is useful in the optical resolution of D,L-pantolactone via D-selective asymmetric hydrolysis and a gene encoding the the same are provided. The invention discloses the gene coding for a natural D-pantolactone hydrolase (for example, one originating in Fusarium oxysporum) or proteins having an activity substantially equivalent thereto; host cells transformed with DNA containing a nucleotide sequence coding for said protein, processes for producing said protein via using said host cells and uses of said proteins and host cells.

    摘要翻译： 提供了可用于通过D-选择性不对称水解的D，L-泛酸内酯的光学拆分和编码其的基因的新型酶。 本发明公开了编码天然D-泛酸内酯水解酶（例如源于尖孢镰刀菌的一种）的基因或具有与其基本相当的活性的蛋白质; 用含有编码所述蛋白质的核苷酸序列的DNA转化的宿主细胞，通过使用所述宿主细胞产生所述蛋白质的过程和所述蛋白质和宿主细胞的用途。

    Media for culturing microorganisms and process for producing unsaturated fatty acids or lipids containing the same
    36.
    发明授权
    Media for culturing microorganisms and process for producing unsaturated fatty acids or lipids containing the same 有权
    用于培养微生物的培养基和用于生产不饱和脂肪酸或含有它们的脂质的方法

    公开(公告)号：US06746857B2

    公开(公告)日：2004-06-08

    申请号：US09331759

    申请日：1999-06-25

    申请人： Kenichi Higashiyama ,  Toshiaki Yaguchi ,  Kengo Akimoto ,  Sakayu Shimizu

    发明人： Kenichi Higashiyama ,  Toshiaki Yaguchi ,  Kengo Akimoto ,  Sakayu Shimizu

    IPC分类号： C12P764

    CPC分类号： C12P7/6427 ,  C12N1/14 ,  C12P7/6472

    摘要： A process for controlling the mycelial morphology of a microorganism belonging to the genus Mortierella during culturing and a process for producing unsaturated fatty acids and a lipid containing them by using a culture medium for culturing a microorganism in which phosphate ions, potassium ions, sodium ions, magnesium ions, and calcium ions in the culture medium are in the range of 5 to 60 mM, 5 to 60 mM, 2 to 50 mM, 0.5 to 9 mM, and 0.5 to 12 mM, respectively, characterized in that the microorganism belonging to the genus Mortierella is cultured in a culture medium containing phosphate ions in the range of 5 to 60 mM, potassium ions in the range of 5 to 60 mM, sodium ions in the range of 2 to 50 mM, magnesium ions in the range of 0.5 to 9 mM, and calcium ions in the range of 0.5 to 12 mM, respectively, to produce unsaturated fatty acids and the lipid containing them, and the culture medium for culturing a microorganism having phosphate ions, potassium ions, sodium ions, magnesium ions, and calcium ions in the range of 5 to 60 mM, 5 to 60 mM, 2 to 50 mM, 0.5 to 9 mM, and 0.5 to 12 mM, respectively.

    摘要翻译： 用于在培养期间控制属于被孢霉属属的微生物的菌丝体形态的方法以及通过使用培养基培养不饱和脂肪酸和含有它们的脂质的方法来培养微生物的方法，其中磷酸根离子，钾离子，钠离子， 培养基中的镁离子和钙离子分别为5〜60mM，5〜60mM，2〜50mM，0.5〜9mM，以及0.5〜12mM的范围，其特征在于，属于 将被孢霉属属菌体培养在含有5-60mM磷酸根离子，5-60mM范围钾离子，2-50mM范围内的钠离子，0.5- 至9mM，钙离子分别为0.5-12mM，以产生不饱和脂肪酸和含有它们的脂质，以及用于培养具有磷酸根离子，钾离子，钠离子，镁离子的微生物的培养基 s和钙离子分别在5至60mM，5至60mM，2至50mM，0.5至9mM和0.5至12mM的范围内。

    Edible oil containing arachidonic acid and foods containing the same
    37.
    发明授权
    Edible oil containing arachidonic acid and foods containing the same 有权
    含有花生四烯酸的食用油和含有它的食品

    公开(公告)号：US6117905A

    公开(公告)日：2000-09-12

    申请号：US284205

    申请日：1999-06-29

    申请人： Kenichi Higashiyama ,  Kengo Akimoto ,  Sakayu Shimizu ,  Nobushige Doisaki ,  Kiyomi Furihata

    发明人： Kenichi Higashiyama ,  Kengo Akimoto ,  Sakayu Shimizu ,  Nobushige Doisaki ,  Kiyomi Furihata

    IPC分类号： A23C9/152 ,  A23D9/00 ,  A23D9/007 ,  A23L1/30 ,  C12P7/64 ,  C12R1/645 ,  A61K31/20

    CPC分类号： A23D9/00 ,  A23L33/12 ,  A23L33/40 ,  A23V2002/00

    摘要： Edible oil containing arachidonic acid obtained from microorganisms belonging to the subgenus Mortierella of the genus Mortierella and being capable of producing arachidonic acid are provided. The oil contain little unsaponifiable matters and, above all, the smallest possible amount of sterol with cyclopropane structure which have not been recognized as food components, and are suitable for the production of foods, in particular, infant formula.Arachidonic acid-containing edible oil originating in microorganisms containing not more than 0.8% by weight, preferably not more than 0.6% by weight of unsaponifiable matters and 20% by weight or more of arachidonic acid. Further, these edible oil contain not more than 0.3% by weight, preferably not more than 0.15% by weight of 24,25-methylenecholest-5-en-3 .beta.-ol. The microorganisms are those belonging to the subgenus Mortierella of the genus Mortierella and being capable of producing arachidonic acid. These microorganisms belong to the species alpine of the genus Mortierella. Foods including the arachidonic acid-containing edible oil. Formula for premature infants, formula for infants, foods for infants, and foods for pregnant women and nursing mothers, including the arachidonic acid-containing edible oil.

    摘要翻译： PCT No.PCT / JP97 / 03631 Sec。 371日期1999年6月29日第 102（e）1999年6月29日PCT PCT 1997年10月9日PCT公布。 公开号WO98 / 16119 日期：1998年04月23日提供了从属于被孢霉属的被孢霉属的属于微生物的微生物获得的能够产生花生四烯酸的花生四烯酸的可食用油。 该油含有少量不皂化物质，最重要的是尽可能少量的具有环丙烷结构的固醇未被认定为食品成分，并且适用于生产食品，特别是婴儿配方奶粉。 来源于不超过0.8重量％，优选不超过0.6重量％的不皂化物和20重量％以上花生四烯酸的微生物的含花生四烯酸的食用油。 此外，这些食用油含有不多于0.3重量％，优选不超过0.15重量％的24,25-亚甲基胆甾-5-烯-3β-醇。 这些微生物属于被孢霉属的被孢霉属亚属属，并且能够产生花生四烯酸。 这些微生物属于被孢霉属的高山属。 食物包括含花生四烯酸的食用油。 婴幼儿配方奶粉，婴儿配方食品，孕妇食品，哺乳母亲，包括含花生四烯酸的食用油。

    D-pantolactone hydrolase and process for the preparation thereof
    38.
    发明授权
    D-pantolactone hydrolase and process for the preparation thereof 失效
    D-泛酸内酯水解酶及其制备方法

    公开(公告)号：US5372940A

    公开(公告)日：1994-12-13

    申请号：US859439

    申请日：1992-06-02

    申请人： Keiji Sakamoto ,  Hideaki Yamada ,  Sakayu Shimizu

    发明人： Keiji Sakamoto ,  Hideaki Yamada ,  Sakayu Shimizu

    IPC分类号： C12N9/18 ,  C12P7/42 ,  C12P17/04 ,  C12P41/00 ,  C12R1/645 ,  C12R1/665 ,  C12R1/77 ,  C12R1/82 ,  C12R1/845 ,  C12P17/02 ,  C12N9/14

    CPC分类号： C12P7/42 ,  C12N9/18 ,  C12P17/04 ,  C12P41/005

    摘要： The invention is drawn to a new enzyme that selectively hydrolyzes D-pantolactone in D,L-pantolactone and has the following characteristics:(a) action: acts on pantolactone to produce the corresponding acid;(b) specificity for substrate: acts specifically on D-pantolactone but not on L-pantolactone;(c) pH stability: stable at pH5-9;(d) optimal pH: 7.0-7.5;(e) optimal temperature: ca. 50.degree. C.; and(f) effect of various metal ions or inhibitors: inhibited by Cd.sup.2+, Hg.sup.2+ or Cu.sup.2+ EDTA, as well as a process for the preparation thereof. The enzyme is preferably produced by cultivating the microorganisms of the genera Fusarium, Cylindrocarpon, and Gibberella, and most preferably the microorganism Fusarium oxysporum IFO 5942.

    摘要翻译： PCT No.PCT / JP91 / 01351 Sec。 371日期：1992年6月2日 102（e）日期1992年6月2日PCT提交1991年10月4日PCT公布。 公开号WO92 / 06182 日期：1992年4月16日。本发明涉及一种在D，L-泛酸内酯中选择性水解D-泛酸内酯的新酶，具有以下特征：（a）作用：作用于泛酸内酯以产生相应的酸; （b）底物的特异性：特异性地对D-泛酸内酯作用，但不对L-泛酸内酯作用; （c）pH稳定性：pH5-9时稳定; （d）最佳pH：7.0-7.5; （e）最佳温度：约 50℃。 和（f）各种金属离子或抑制剂的作用：被Cd2 +，Hg2 +或Cu2 + EDTA抑制，以及其制备方法。 该酶优选通过栽培镰刀菌属，圆锥菌属和赤霉属的微生物，最优选微生物尖孢镰刀菌IFO 5942来产生。

    Process for preparing acyl-CoA synthetase LCF-18
    40.
    发明授权
    Process for preparing acyl-CoA synthetase LCF-18 失效
    制备酰基辅酶A合成酶LCF-18的方法

    公开(公告)号：US4229538A

    公开(公告)日：1980-10-21

    申请号：US28604

    申请日：1979-04-09

    申请人： Hideaki Yamada ,  Sakayu Shimizu ,  Yoshiki Tani

    发明人： Hideaki Yamada ,  Sakayu Shimizu ,  Yoshiki Tani

    IPC分类号： C12N9/00 ,  C12Q1/25 ,  C12N9/10

    CPC分类号： C12N9/93 ,  C12Q1/25 ,  Y10S435/875 ,  Y10S435/911

    摘要： Acyl-CoA synthetase, having a high activity to C.sub.16 -C.sub.18 long chain fatty acids, is obtained by culturing strains belonging to various genera. As is known, acyl-CoA synthetase, having a strong activity to C.sub.16 -C.sub.18 long chain fatty acids, is generally obtained from liver of rat; however, it has now been discovered that acyl-CoA synthetase can be obtained from microorganisms and this enzyme is called acyl-CoA synthetase LCF-18. By use of acyl-CoA synthetase LCF-18 of the present invention which has a high activity to C.sub.16 -C.sub.18 long chain fatty acids, serum non-esterified fatty acid of human beings can be accurately determined, and it is very useful for diagnosis of diabetes and so forth.

    摘要翻译： 通过培养属于各种属的菌株获得对C16-C18长链脂肪酸具有高活性的酰基辅酶A合成酶。 众所周知，对C16-C18长链脂肪酸具有强烈活性的酰基辅酶A合成酶通常由大鼠肝脏获得; 然而，现在已经发现可以从微生物获得酰基辅酶A合成酶，该酶称为酰基辅酶A合成酶LCF-18。 通过使用对C16-C18长链脂肪酸具有高活性的本发明的酰基-CoA合成酶LCF-18，可以准确测定人的血清非酯化脂肪酸，并且其对于诊断 糖尿病等。