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    Production of biotin
    12.
    发明授权
    Production of biotin 失效
    生物素生产

    公开(公告)号：US06361978B1

    公开(公告)日：2002-03-26

    申请号：US08840059

    申请日：1997-04-24

    申请人： Tatsuo Hoshino ,  Akira Asakura ,  Tatsuya Kiyasu ,  Yoshie Nagahashi

    发明人： Tatsuo Hoshino ,  Akira Asakura ,  Tatsuya Kiyasu ,  Yoshie Nagahashi

    IPC分类号： C12P1718

    CPC分类号： C12N9/93 ,  C12N9/13 ,  C12P17/186

    摘要： The present invention provides processes for making biotin from desthiobiotin by either contacting desthiobiotin with an enzyme reaction mixture containing bioB gene product and nifU gene product and/or nifS gene product and isolating the biotin or cultivating a microorganism transformed with DNA encoding the bioB gene product, nifU gene product and nifS gene product and isolating the biotin.

    摘要翻译： 本发明提供通过将脱硫生物素与含有bioB基因产物和nifU基因产物和/或nifS基因产物的酶反应混合物接触来生产脱氧生物素的生物素的方法，并分离生物素或培养用编码bioB基因产物的DNA转化的微生物， nifU基因产物和nifS基因产物，并分离生物素。

    Process of producing antibiotic SM-173B with Streptomyces chromofuscus
    16.
    发明授权
    Process of producing antibiotic SM-173B with Streptomyces chromofuscus 失效
    生产抗生素SM-173B与链霉菌（Streptomyces chromofuscus）的方法

    公开(公告)号：US4304861A

    公开(公告)日：1981-12-08

    申请号：US92402

    申请日：1979-11-08

    申请人： Akiko Fujiwara ,  Mitsuhiko Fujiwara ,  Tatsuo Hoshino ,  Yuzuru Sekine ,  Masaaki Tazoe

    发明人： Akiko Fujiwara ,  Mitsuhiko Fujiwara ,  Tatsuo Hoshino ,  Yuzuru Sekine ,  Masaaki Tazoe

    IPC分类号： C12P29/00 ,  A61K31/65 ,  A61P31/04 ,  C07G11/00 ,  C12P15/00

    CPC分类号： C07G11/00 ,  Y10S435/886

    摘要： A novel antibiotic having the ascribed formula ##STR1## is provided. They physical characteristics are as follows: (a) Elementary analysis: Calculated (for C.sub.20 H.sub.16 O.sub.7): C,65.22; H,4.35%. Found: C,64.60; H,4.34%.(b) Molecular weight (mass-spectrometric): 368.(c) Melting point: 225.degree.-226.degree. C.(d) Specific rotation: [.alpha.].sub.D .sup.20 =+123.5.degree. (c=0.2 in chloroform).(e) Ultraviolet absorption spectrum:.lambda..sub.max .sup.methanol =235 nm (.epsilon.=28600); 262 nm (.epsilon.=21900); 434-435 nm (.epsilon.=11300);.lambda..sub.max .sup.0.1-N NaOH/methanol =246 nm (.epsilon.=22400); 256 nm (.epsilon.=23200); 523-524 nm (.epsilon.=9600).(f) Infrared absorption spectrum: 3500, 1710, 1672, 1628, 1600, 1575, 1472, 1452, 1420, 1390, 1295, 1255, 1218, 1200, 1182, 1105, 1025, 1005, 962, 938, 848, 765, 752, 730 cm.sup.-1.(g) Solubility: Soluble in methanol, ethanol, butanol, acetone, chloroform, ethyl acetate and in water under alkaline conditions; poorly soluble in n-hexane; insoluble in petroleum ether and water.(h) Color reactions: Positive with sulfuric acid and iodine; negative with ninhydrin and anisaldehyde.(i) NMR spectrum in DC.Cl.sub.3 : 1.42 ppm (3H, singlet), 3.02 ppm (1H, quartet), 3.43 ppm (3H, singlet), 3.62 ppm (1H, triplet), 3.64 ppm (1H, singlet), 3.93 ppm (1H, triplet), 7.31-3.41 ppm (4H, olefinic protons), 12.0 ppm (1H, singlet), 12.5 ppm (1H, singlet).Also provided is a process for the antibiotic's production.

    摘要翻译： 提供具有归属公式的新型抗生素。 它们的物理特性如下：（a）元素分析：计算值（C 20 H 16 O 7）：C，65.22; H，4.35％。 实测值：C，64.60; H，4.34％。 （b）分子量（质谱）：368.（c）熔点：225°-226℃。（d）比旋光度：αD20 = + 123.5°（氯仿中c = 0.2）。 （e）紫外吸收光谱：λmax甲醇= 235nm（ε= 28600）; 262nm（ε= 21900）; 434-435nm（ε= 11300）; λmax 0.1-N NaOH /甲醇= 246nm（ε= 22400）; 256nm（ε= 23200）; 523-524nm（ε= 9600）。 （f）红外吸收光谱：3500,1710,1672,1628,1600,1575,1472,1452,1420,1390,1295,1255,1218,1200,1122,1105,1025,1005，962，938,848,765 ，752,730cm-1。 （g）溶解性：在碱性条件下，溶于甲醇，乙醇，丁醇，丙酮，氯仿，乙酸乙酯和水; 难溶于正己烷; 不溶于石油醚和水。 （h）显色反应：用硫酸和碘阳性; 用茚三酮和茴香醛阴性。 （i）DC中的NMR光谱：3.43ppm（3H，单峰），3.02ppm（1H，四重峰），3.43ppm（3H，单峰），3.62ppm（1H，三重峰） 3.93ppm（1H，三重峰），7.31-3.41ppm（4H，烯属质子），12.0ppm（1H，单峰），12.5ppm（1H，单峰）。 还提供了抗生素生产的过程。

    Genetically engineered L-sorbose reductase-deficient mutants
    17.
    发明授权
    Genetically engineered L-sorbose reductase-deficient mutants 有权
    遗传工程L-山梨糖还原酶缺陷突变体

    公开(公告)号：US06664082B1

    公开(公告)日：2003-12-16

    申请号：US09266320

    申请日：1999-03-11

    申请人： Tatsuo Hoshino ,  Takahide Kon ,  Masako Shinjoh ,  Masaaki Tazoe

    发明人： Tatsuo Hoshino ,  Takahide Kon ,  Masako Shinjoh ,  Masaaki Tazoe

    IPC分类号： C12P1902

    CPC分类号： C12N9/0004 ,  C12N15/102

    摘要： The present invention provides a genetically engineered microorganism belonging to the genus Gluconobacter or Acetobacter, which has an engineered gene for the biological activity of reducing L-sorbose which is more than 90% non-functional in developing the said biological activity. The engineered microorganism is derived from a microorganism belonging to the genus Gluconobacter or Acetobacter, and has the biological activity for reducing L-sorbose that is less than 10% of the amount of the activity of the wild type organism. The present invention further provides a method for producing the genetically engineered microorganism by mutating the gene of a microorganism of the genus Gluconobacter or Acetobacter which encodes a protein having the L-sorbose reductase activity, determining the L-sorbose reductase activity of the resulting microorganism with the mutated gene and selecting the genetically engineered microorganism having L-sorbose reductase activity that is less than 10% of the amount of the activity of the wild type microorganism.

    摘要翻译： 本发明提供属于葡萄糖酸杆菌属或醋杆菌属的遗传工程微生物，其具有用于降低L-山梨糖的生物活性的工程化基因，其在发展所述生物活性时超过90％不起作用。 工程微生物来自属于葡糖杆菌属或醋杆菌属的微生物，具有小于野生型生物活性量的10％的L-山梨糖的生物活性。 本发明还提供了通过使编码具有L-山梨糖还原酶活性的蛋白质的葡糖酸杆菌属或醋酸杆菌属的微生物的基因突变来生产遗传工程微生物的方法，用所得的微生物测定所得微生物的L-山梨糖还原酶活性 突变基因并选择具有小于野生型微生物活性量的10％的L-山梨糖还原酶活性的遗传工程微生物。

    Production of vitamin B.sub.6 with an enzyme-containing cell extract
    18.
    发明授权
    Production of vitamin B.sub.6 with an enzyme-containing cell extract 失效
    用含酶细胞提取物生产维生素B6

    公开(公告)号：US6060267A

    公开(公告)日：2000-05-09

    申请号：US291718

    申请日：1999-04-14

    申请人： Tatsuo Hoshino ,  Masaaki Tazoe

    发明人： Tatsuo Hoshino ,  Masaaki Tazoe

    IPC分类号： C12P17/12 ,  A23L1/30 ,  A61K51/00 ,  C12N1/00 ,  C12P1/00 ,  C12P1/04

    CPC分类号： C12P17/12 ,  Y10S435/822

    摘要： A process for the enzymatic production of vitamin B.sub.6 which includes incubating 1-deoxy-D-threo-pentulose and 4-hydroxy-L-threonine with an enzyme system that is cell free or essentially cell free and prepared from the cells of a microorganism belonging to the genus Rhizobium, Sinorhizobium, Flavobacterium, Chryseobacterium, Lactobacillus, Arthrobacter, Bacillus, Klebsiella, Escherichia, Pseudomonas, Stenotrophomonas, Enterobacter, Serratia, Corynebacterium, Brevibacterium, Exiguobacterium, Saccharomyces, Yamadazyma, Pichia or Candida, in the presence of NADP.sup.+, NAD.sup.+, ATP. Manganese and magnesium ions stimulate the above reaction. This process affords high yields of vitamin B.sub.6, a vitamin essential for the nutrition of animals, plants and microorganisms, and which is also useful as a medicine or food additive. In addition, an enzyme reaction system for producing vitamin B.sub.6 and a process for making the enzyme reaction system are also provided.

    摘要翻译： 用于酶促生产维生素B6的方法，其包括用无细胞或基本上不含细胞的酶系统温育1-脱氧-D-苏糖 - 戊酮糖和4-羟基-L-苏氨酸并由所属微生物的细胞制备 在NADP +，NAD +的存在下，根瘤菌属，根瘤菌属，中华根瘤菌属，黄杆菌属，巧克力杆菌属，乳杆菌属，节杆菌属，芽孢杆菌属，克雷伯菌属，埃希氏杆菌属，假单胞菌属，嗜麦芽寡养单胞菌属，肠杆菌属，沙雷氏菌属，棒状杆菌属，短杆菌属，精氨酸杆菌属，酵母属，Yamadazyma，毕赤酵母或假丝酵母属， ，ATP。 锰和镁离子刺激上述反应。 该方法提供高产量的维生素B6，维生素B6是动物，植物和微生物营养必需的维生素，也可用作药物或食品添加剂。 此外，还提供了用于生产维生素B6的酶反应体系和制备酶反应体系的方法。

    Process for the production of d-biotin
    19.
    发明授权
    Process for the production of d-biotin 失效
    生产生物素的方法

    公开(公告)号：US5922581A

    公开(公告)日：1999-07-13

    申请号：US833411

    申请日：1997-04-04

    申请人： Tatsuo Hoshino ,  Akifumi Noro ,  Masaaki Tazoe

    发明人： Tatsuo Hoshino ,  Akifumi Noro ,  Masaaki Tazoe

    IPC分类号： C12N1/20 ,  C12P17/18 ,  C12R1/01 ,  C12N1/00 ,  C12P17/10

    CPC分类号： C12P17/186 ,  Y10S435/822

    摘要： A process for producing d-biotin comprises cultivating a microorganism of the genus Kurthia and which is resistant to biotin antimetabolites and capable of producing d-biotin in a medium under aerobic conditions, and separating the resulting d-biotin from the fermentation broth. The cultivation medium suitably contains an assimilable carbon source, a digestible nitrogen source, inorganic salts and other nutrients necessary for the growth of the microorganism at a pH of about 5-9, temperature of about 10-40.degree. C. and for a duration of about 1-10 days. The preferred microorganism are Kurthia sp. 538-KA26, 538-17H4, 538-51F9 and 538-2A 13 (DSM No. 10609, 10608, 10610 and 10607, respectively), which are also new, and as such represent a further aspect of the present invention. The so-produced d-biotin is one of the essential vitamins for the nutrition of animals, plants and microorganisms, and is important as a medicine or food additive. It is produced in particularly high yield by the process of the present invention.

    摘要翻译： 一种生产d-生物素的方法包括培养Kurthia属的微生物，其对生物素抗代谢物具有抗性，并且能够在需氧条件下在培养基中产生d-生物素，并将所得的d-生物素与发酵液分离。 栽培培养基适当地含有可吸收的碳源，可消化的氮源，无机盐和微生物生长所需的其它营养物，pH约为5-9，温度约为10-40℃，持续时间为 约1-10天。 优选的微生物是Kurthia sp。 538-KA26,538-17H4,538-51F9和538-2A 13（分别为DSM10609,10608,101010和10607），它们也是新的，因此代表本发明的另一方面。 如此生产的d生物素是动物，植物和微生物营养的必需维生素之一，并且作为药物或食品添加剂是重要的。 通过本发明的方法产生的产率特别高。