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1. 发明授权

US06303349B1 Esterase and methods for the production of optically active chroman compounds 失效
标题翻译：酯酶和用于制备光学活性色满化合物的方法
公开(公告)号：US06303349B1
公开(公告)日：2001-10-16
申请号：US09524760
申请日：2000-03-13
申请人： Toshihiro Oikawa , Nobuhiro Fukuhara , Yasuko Matsuba
发明人： Toshihiro Oikawa , Nobuhiro Fukuhara , Yasuko Matsuba
IPC分类号： C12P1706
CPC分类号： C12N9/18 , C12P17/06 , C12P41/005
摘要： Optically active chroman-3-acetic acids and optically active chroman-3-acetic acid esters can be obtained by treating a mixture of (3R)- and (3S)-chroman-3-acetic esters of formula (I) [wherein R1 is a straight or branched alkyl group having 1-5 carbon atoms and R2 is a hydrogen atom or substituted or unsubstituted amino group] is treated with an esterase which has an optically selective hydrolyzing activity or microorganisms which carry said hydrolase, or a preparation therefrom. A novel esterase derived from bacteria of genus Pseudonocardia can also be used as said esterase.
摘要翻译：通过处理式（I）的（3R） - （3S） - 苯并二氢吡喃-3-乙酸酯的混合物[其中R 1是（R 3））和（3S） - 苯并二氢吡喃-3-乙酸酯的混合物可以获得光学活性色满-3-乙酸和光学活性色满-3-乙酸酯具有1-5个碳原子的直链或支链烷基，R2是氢原子或取代或未取代的氨基]用具有光学选择性水解活性的酯酶或携带所述水解酶的微生物或其制备物进行处理。也可以使用衍生自假诺卡氏菌属细菌的新型酯酶作为所述酯酶。

2. 发明授权

US6060290A Esterase and methods for the production of optically active chroman compounds 失效
标题翻译：酯酶和用于制备光学活性色满化合物的方法
公开(公告)号：US6060290A
公开(公告)日：2000-05-09
申请号：US118275
申请日：1998-07-17
申请人： Toshihiro Oikawa , Nobuhiro Fukuhara , Yasuko Matsuba
发明人： Toshihiro Oikawa , Nobuhiro Fukuhara , Yasuko Matsuba
IPC分类号： C12N9/18 , C12N15/55 , C12P17/06 , C12P41/00
CPC分类号： C12N9/18 , C12P17/06 , C12P41/005
摘要： Optically active chroman-3-acetic acids and optically active chroman-3-acetic acid esters can be obtained by treating a mixture of (3R)- and (3S)-chroman-3-acetic esters of formula (I) ##STR1## [wherein R.sub.1 is a straight or branched alkyl group having 1-5 carbon atoms and R.sub.2 is a hydrogen atom or substituted or unsubstituted amino group]is treated with an esterase which has an optically selective hydrolyzing activity or microorganisms which carry said hydrolase, or a preparation therefrom. A novel esterase derived from bacteria of genus Pseudonocardia can also be used as said esterase.
摘要翻译：通过处理式（I）的（3R） - （3S） - 苯并二氢吡喃-3-乙酸酯的混合物[其中R 1是（R 3））和（3S） - 苯并二氢吡喃-3-乙酸酯的混合物可以获得光学活性色满-3-乙酸和光学活性色满-3-乙酸酯具有1-5个碳原子的直链或支链烷基，R2是氢原子或取代或未取代的氨基]用具有光学选择性水解活性的酯酶或携带所述水解酶的微生物或其制备物进行处理。也可以使用衍生自假诺卡氏菌属细菌的新型酯酶作为所述酯酶。

3. 发明授权

US5382517A Process for the preparation of L-serine by an enzymatic method 失效
标题翻译：通过酶法制备L-丝氨酸的方法
公开(公告)号：US5382517A
公开(公告)日：1995-01-17
申请号：US20446
申请日：1993-02-22
申请人： Daisuke Ura , Tadashi Hashimukai , Toshio Matsumoto , Nobuhiro Fukuhara
发明人： Daisuke Ura , Tadashi Hashimukai , Toshio Matsumoto , Nobuhiro Fukuhara
IPC分类号： C12P13/06 , C12R1/19
CPC分类号： C12P13/06
摘要： The invention relates to a process for preparing L-serine from glycine and formaldehyde in the presence of cells of a microorganism or a cell-treated product having a serine hydroxymethyl transferase activity by an enzymatic method.
摘要翻译：本发明涉及通过酶法在微生物细胞或具有丝氨酸羟甲基转移酶活性的细胞处理产物存在下从甘氨酸和甲醛制备L-丝氨酸的方法。

4. 发明授权

US4946790A Recombinant plasmid for the expression of L-phenylalanine ammonia-lyase and transformed strain carrying same 失效
标题翻译：用于表达L-苯丙氨酸氨裂解酶和携带相同转化菌株的重组质粒
公开(公告)号：US4946790A
公开(公告)日：1990-08-07
申请号：US151234
申请日：1988-02-01
申请人： Nobuhiro Fukuhara , Setsuo Yoshino , Satori Sone , Yoshiyuki Nakajima , Nobuyoshi Makiguchi
发明人： Nobuhiro Fukuhara , Setsuo Yoshino , Satori Sone , Yoshiyuki Nakajima , Nobuyoshi Makiguchi
IPC分类号： C12N1/21 , C12N9/88 , C12N15/70
CPC分类号： C12N9/88 , C12N15/70
摘要： A recombinant plasmid for the expression of phenylalanine ammonia-lyase (PAL) is constructed by incorporating therein a combined promoter comprising (a) the fusion promoter (the tac promoter) composed of the trp promoter minus 35 region and the lac UV-5 promoter minus 10 region and (b) the P.sub.L promoter of the lambda phage, the tac promoter and the P.sub.L promoter being connected so as to have the same directional property. This recombinant plasmid permits more efficient expression of PAL in Escherichia coli.

5. 发明授权

US06333427B1 Process for preparing N-t-butoxycarbonylphenylalanine ester 失效
标题翻译：制备N-叔丁氧基羰基苯丙氨酸酯的方法
公开(公告)号：US06333427B1
公开(公告)日：2001-12-25
申请号：US09507676
申请日：2000-02-22
申请人： Hitoki Miyake , Masami Osabe , Mitsuo Koito , Setsuo Yoshino , Nobuhiro Fukuhara
发明人： Hitoki Miyake , Masami Osabe , Mitsuo Koito , Setsuo Yoshino , Nobuhiro Fukuhara
IPC分类号： C07C22900
CPC分类号： C07C269/08 , C07B2200/07 , C07C269/04 , C07C271/22
摘要： A purified N-t-butoxycarbonylphenylalanine ester preparation having an enhanced optical activity can be obtained by bringing an N-t-butoxycarbonylphenylalanine ester preparation containing an optically active compound into contact with an aliphatic hydrocarbon, extracting the optically active compound with the aliphatic hydrocarbon, and recovering the optically active compound from the resulting extract.
摘要翻译：具有增强的光学活性的纯化的N-叔丁氧基羰基苯丙氨酸酯制剂可以通过使含有光学活性化合物的N-叔丁氧基羰基苯丙氨酸酯制剂与脂族烃接触，用脂族烃萃取旋光活性化合物，并回收光学活性从所得提取物中加入化合物。

6. 发明授权

US5312977A Method for purifying L-phenylalanine 失效
标题翻译： L-苯丙氨酸的纯化方法
公开(公告)号：US5312977A
公开(公告)日：1994-05-17
申请号：US19116
申请日：1993-02-17
申请人： Makoto Yanaka , Tooru Miyahara , Daisuke Ura , Nobuhiro Fukuhara
发明人： Makoto Yanaka , Tooru Miyahara , Daisuke Ura , Nobuhiro Fukuhara
IPC分类号： C07C227/42 , C12P13/22 , C07C229/28
CPC分类号： C07C227/42 , C12P13/222
摘要： L-Phenylalanine is isolated in pure form from an aqueous solution thereof containing cinnamic acid by subjecting the aqueous solution to toluene extraction to extract cinnamic acid therefrom; separating the toluene phase; and then concentrating the aqueous phase, until crystals of L-phenylalanine form therein, while concurrently separating the crystalline L-phenylalanine from the liquid aqueous phase.
摘要翻译：通过使水溶液进行甲苯萃取从其中提取肉桂酸，从含有肉桂酸的水溶液中分离出纯净的L-苯丙氨酸，分离甲苯相; 然后浓缩水相，直到形成L-苯丙氨酸晶体，同时从液体水相中同时分离结晶L-苯丙氨酸。

7. 发明授权

US5322786A Method of regulating expression of a foreign gene by controlling culture temperature and a process of producing a foreign gene product thereby 失效
标题翻译：通过控制培养温度调节外源基因表达的方法和由此产生外源基因产物的方法
公开(公告)号：US5322786A
公开(公告)日：1994-06-21
申请号：US798044
申请日：1991-11-27
申请人： Nobuhiro Fukuhara , Setsuo Yoshino , Kaoru Yamamoto , Satori Sone , Maki Suzuki , Yoshiyuki Nakajima
发明人： Nobuhiro Fukuhara , Setsuo Yoshino , Kaoru Yamamoto , Satori Sone , Maki Suzuki , Yoshiyuki Nakajima
IPC分类号： C12N9/88 , C12N15/60 , C12N15/70 , C12N1/21 , C12N15/00 , C12N15/63
CPC分类号： C12N15/70 , C12N9/88
摘要： Escherichia coil carrying a hybrid plasmid having been constructed by inserting a desired foreign gene into an expression vector so as to permitting expression of said desired foreign gene therein was cultured at a temperature 40.degree. C. or over so that the expression of said desired foreign gene was suppressed. This E. coil (i.e., transformant) was cultured at 40.degree. C. or over in a first process to suppress the expression of the foreign gene and to support sufficient cell growth and thereafter below 40.degree. C. in a second process to release the suppression of the expression so as to permit effective production of the foreign gene product, which resulted in high concentration of the foreign gene product in the final culture.
摘要翻译：携带通过将期望的外源基因插入表达载体以便允许其中表达所需外源基因的杂交质粒的大肠杆菌在40℃或更高的温度下培养，使得所需外源基因的表达被压制在40℃或以上培养该E.线圈（即转化体）的第一过程中，抑制外源基因的表达并支持足够的细胞生长，然后在第二过程中低于40℃释放抑制表达以允许有效产生外源基因产物，这导致外源基因产物在最终培养物中的高浓度。

8. 发明授权

US5139935A Method of regulating expression of a foreign gene by controlling the sugar concentration in a medium and a process of producing a foreign product thereby 失效
标题翻译：通过控制培养基中的糖浓度和由此制造异物的方法来调节外源基因的表达的方法
公开(公告)号：US5139935A
公开(公告)日：1992-08-18
申请号：US659472
申请日：1991-02-25
申请人： Nobuhiro Fukuhara , Setsuo Yoshino , Satori Sone , Yoshiyuki Nakajima , Nobuyoshi Makiguchi
发明人： Nobuhiro Fukuhara , Setsuo Yoshino , Satori Sone , Yoshiyuki Nakajima , Nobuyoshi Makiguchi
IPC分类号： C12N1/21 , C12N9/88 , C12N15/60 , C12N15/70
CPC分类号： C12N15/70 , C12N9/88
摘要： Escherichia coli carrying a hybrid plasmid having been constructed by inserting a desired foreign gene into an expression vector so as to permitting expression of said desired foreign gene therein was cultured in a medium containing a sugar component utilizable by the E. coli as a carbon source at a concentration of 0.3% or more so that the expression of said desired foreign gene was suppressed. This E. coli (i.e., transformant) was cultured in the medium in which the sugar concentration was maintained at 0.3% or more in a first process so as to supress the suppression of the foreign gene and to support sufficient cell growth and thereafter at less than 0.3% in a second process to release the suppression of the expression so as to permit effective production of the foreign gene product, which resulted in high concentration of the foreign gene product in the final culture.
摘要翻译：携带通过将期望的外源基因插入到表达载体中以便允许其中表达所需外源基因的杂交质粒的大肠杆菌在含有作为碳源的大肠杆菌可利用的糖成分的培养基中培养浓度为0.3％以上，能够抑制所希望的外源基因的表达。将该大肠杆菌（即转化体）在第一种方法中将糖浓度保持在0.3％以上的培养基中培养，以抑制外源基因的抑制并支持足够的细胞生长，此后在少于在第二过程中超过0.3％以释放表达的抑制，以便有效地产生外源基因产物，这导致外源基因产物在最终培养物中的高浓度。

9. 发明授权

US06472196B1 Amino acid sequence of L-phenylalanine ammonia-lyase 失效
标题翻译： L-苯丙氨酸氨裂解酶的氨基酸序列
公开(公告)号：US06472196B1
公开(公告)日：2002-10-29
申请号：US08214018
申请日：1994-03-15
申请人： Nobuhiro Fukuhara , Setsuo Yoshino , Kaoru Yamamoto , Tomoyuki Se , Satori Sone , Yoshiyuki Nakajima , Maki Suzuki , Nobuyoshi Makiguchi
发明人： Nobuhiro Fukuhara , Setsuo Yoshino , Kaoru Yamamoto , Tomoyuki Se , Satori Sone , Yoshiyuki Nakajima , Maki Suzuki , Nobuyoshi Makiguchi
IPC分类号： C12N988
CPC分类号： C12P13/222 , C12N9/88
摘要： In order to provide technical information necessary for the production of L-phenylalanine ammonia-lyase (PAL) by utilizing genetic engineering techniques, the structural gene for PAL and the amino acid sequence of PAL have been elucidated in Rhodosporidium toruloides, and novel recombinant DNA plasmids (e.g., pSW101, pYtrp6 and pKY201) have been created by inserting a DNA strand coding for the PAL gene between the 3′-terminus of the promoter region and the 5′-terminus of the terminator region. Moreover, transformants having such a novel recombinant DNA plasmid have been created, and a process for the production of PAL by growing such a novel transformant so as to cause PAL to be produced and accumulated in the culture has been established. Furthermore, there has been established a novel technique for the production of L-phenylalanine by reacting an ammonia donor with cinnamic acid in the presence of the PAL prepared by the aforesaid novel process.
摘要翻译：为了提供利用遗传工程技术生产L-苯丙氨酸氨裂解酶（PAL）所必需的技术信息，已经在Rhodosporidium toruloides中阐明了PAL的结构基因和PAL的氨基酸序列，新的重组DNA质粒（例如，pSW101，pYtrp6和pKY201）是通过将编码PAL基因的DNA链插入到启动子区域的3'端和终止区域的5'末端而产生的。此外，具有这样一种新颖的转化体已经建立了重组DNA质粒，并且通过生长这样一种新的转化体以使培养中产生和积累PAL来生产PAL的方法已被建立。此外，已经建立了一种生产新技术在通过上述新方法制备的PAL存在下，使氨供体与肉桂酸反应，生成L-苯丙氨酸。

10. 发明授权

US5756319A Production process of S-phenyl-L-cysteine 失效
标题翻译： S-苯基-L-半胱氨酸的生产工艺
公开(公告)号：US5756319A
公开(公告)日：1998-05-26
申请号：US676666
申请日：1996-07-10
申请人： Kazuhiro Fukuta , Nobuhiro Fukuhara
发明人： Kazuhiro Fukuta , Nobuhiro Fukuhara
IPC分类号： C12P13/12 , C12P13/04 , C12N9/88
CPC分类号： C12P13/12
摘要： S-phenyl-L-cysteine can be produced in a high yield by reacting thiophenol and L-serine under the action of tryptophan synthase at a pH in a range of from 9.0 to 10.5. Purification of S-phenyl-L-cysteine obtained by this enzyme reaction can be effectively achieved by adjusting the pH of a crystal-containing reaction mixture to a strongly acidic level of 1.5 or lower to dissolve crystals of S-phenyl-L-cysteine, adding activated carbon to the pH-adjusted mixture, maintaining the resultant mixture at a temperature of from 20.degree. to 60.degree. C. under aeration with an oxygen-containing gas, subjecting the thus-obtained mixture to filtration, raising the pH of the resulting filtrate back to a range of from 2.5 to 6.0 to precipitate crystals of S-phenyl-L-cysteine and then recovering the thus-precipitated crystals.
摘要翻译： S-苯基-L-半胱氨酸可以通过在色氨酸合成酶的作用下，在9.0至10.5的pH范围内使苯硫酚和L-丝氨酸反应而以高产率制备。通过将含有结晶的反应混合物的pH调节至1.5以下的强酸性水平，可以有效地实现通过该酶反应得到的S-苯基-L-半胱氨酸的纯化，以溶解S-苯基-L-半胱氨酸的结晶，将活性炭加入到pH调节的混合物中，在含氧气体通气下将所得混合物保持在20℃至60℃的温度，对由此得到的混合物进行过滤，使所得混合物的pH值升高滤液回到2.5至6.0的范围，以沉淀S-苯基-L-半胱氨酸的晶体，然后回收如此沉淀的晶体。

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