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1. 发明授权

US5155040A Process for reducing the salt concentration in a biomass suspension 失效
标题翻译：降低生物质悬浮液中盐浓度的方法
公开(公告)号：US5155040A
公开(公告)日：1992-10-13
申请号：US493122
申请日：1990-03-15
申请人： Maria-Regina Kula , Arend Greve
发明人： Maria-Regina Kula , Arend Greve
IPC分类号： B01D11/04 , C07K1/14 , C12N1/00 , C12N9/00 , C12P21/00
CPC分类号： C12N1/005
摘要： Alcohol is suitable for the extraction of salt from biomass-containing suspensions, in particular from the salt-containing lower phase of an aqueous 2-phase extraction of intracellular proteins after cell disruption, with the formation of a salt-containing alcoholic upper phase. It is possible to use ethanol, propanol, isopropanol or tert.-butanol, but especially ethanol, as the alcohol. The upper phase is separated from the lower phase by disk separators or decanters. The extraction is carried out, in particular, as a countercurrent extraction in at least three stages, and uses 10 to 30% by weight salt-containing suspension or liquid with 30 to 50% by weight alcohol, in particular ethanol, remainder water. The alcohol is removed from the resulting salt-rich upper phase by evaporation, and the salt solution is recycled where appropriate after further concentration to obtain proteins.
摘要翻译：醇适用于从含有生物质的悬浮液中提取盐，特别是从细胞破碎后的细胞内蛋白水相2相提取的含盐下层中提取盐，形成含盐醇的上层相。可以使用乙醇，丙醇，异丙醇或叔丁醇，特别是乙醇作为醇。上层通过盘式分离器或倾析器与下层相分离。提取特别是作为至少三个阶段的逆流萃取，并且使用10至30重量％的含盐悬浮液或液体，其中含有30至50重量％的醇，特别是乙醇，剩余的水。通过蒸发从所得的富含盐的上层中除去醇，并且在进一步浓缩后适当地再循环盐溶液以获得蛋白质。

2. 发明申请

US20100009434A1 Method for the Selective Enrichment of Double-Stranded Dna from Nucleic Acid Mixtures 审中-公开
标题翻译：从核酸混合物选择性富集双链Dna的方法
公开(公告)号：US20100009434A1
公开(公告)日：2010-01-14
申请号：US11991947
申请日：2006-09-13
申请人： Markus Müller , Maria-Regina Kula , Jürgen Hubbuch , Andreas Frerix
发明人： Markus Müller , Maria-Regina Kula , Jürgen Hubbuch , Andreas Frerix
IPC分类号： C12N15/00 , C07H1/00
CPC分类号： C12N15/1003
摘要： The invention relates to a method for stripping undesired nucleic acid components from double-stranded DNA, in particular, super-coiled plasmid DNA. The method according to the invention is characterised by the steps: (a) provision of a mixture containing completely and/or partly double-stranded nucleic acids and optionally single-stranded nucleic acids; (b) resuspension of the mixture from step (a) in an aqueous, low-molarity buffer system with low ionic strength and low buffer effect; (c) adjusting conditions in the mixture from step (b), under which the completely and/or partly double-stranded nucleic acids are denatured; (d) further addition of buffer and a polymer component to the mixture from step (c); (e) incubation of the mixture from step (d) for a time which is sufficient for the formation of an aqueous two-phase system with an upper and lower phase; and (f) removal of the upper phase containing the single-strand nucleic acid and collection of the double-strand nucleic acid from the lower phase.
摘要翻译：本发明涉及一种从双链DNA，特别是超螺旋质粒DNA中去除不想要的核酸成分的方法。根据本发明的方法的特征在于以下步骤：（a）提供含有完全和/或部分双链核酸和任选的单链核酸的混合物; （b）将混合物从步骤（a）重悬于具有低离子强度和低缓冲效应的低摩尔浓度缓冲体系中; （c）调节来自步骤（b）的混合物中的完全和/或部分双链核酸变性的条件; （d）进一步向步骤（c）的混合物中加入缓冲剂和聚合物组分; （e）将来自步骤（d）的混合物温育足以形成具有上部和下部相的水相两相体系的时间; 和（f）除去含有单链核酸的上层和从下相收集双链核酸。

3. 发明授权

US06767725B2 Acetyl amino acid racemase from amycolatopsis orientalis for racemizing carbamoyl amino acids 失效
标题翻译：用于外消旋化氨基甲酰氨基酸的乙酰氨基酸消旋酶
公开(公告)号：US06767725B2
公开(公告)日：2004-07-27
申请号：US09973712
申请日：2001-10-11
申请人： Andreas Bommarius , Karlheinz Drauz , Stefan Verseck , Maria-Regina Kula
发明人： Andreas Bommarius , Karlheinz Drauz , Stefan Verseck , Maria-Regina Kula
IPC分类号： C12P1304
CPC分类号： C12N9/90 , C12P41/009
摘要： The invention relates to the use of the N-acetyl amino acid racemase from Amycolatopsis orientalis subspecies lurida for the racemiszation of N-carbamoyl amino acids. This use permits the 100% preparation of optically pure amino acids starting from racemic hydantoins in an enzymatic overall process.
摘要翻译：本发明涉及来自山莨菪属亚种的亚乙酰氨基酸消旋酶用于N-氨基甲酰氨基酸的外消旋化的用途。这种使用允许在酶的总体方法中从外消旋乙内酰脲开始100％制备光学纯氨基酸。

4. 发明授权

US5854035A Enzyme with leuDH activity, nucleotide sequence coding therefor and process for the prepartion of the enzyme 失效
标题翻译：具有leuDH活性的酶，其编码的核苷酸序列和酶的制备过程
公开(公告)号：US5854035A
公开(公告)日：1998-12-29
申请号：US804699
申请日：1997-02-21
申请人： Tanja Stoyan , Maria-Regina Kula
发明人： Tanja Stoyan , Maria-Regina Kula
IPC分类号： C12N15/09 , C07H21/04 , C12N1/21 , C12N9/04 , C12N9/06 , C12N15/00 , C12N15/53 , C12P13/04 , C12P13/06 , C12R1/085 , C12R1/19 , C12N1/00
CPC分类号： C12N9/0016 , Y10S435/834
摘要： The invention relates to an enzyme with LeuDH activity, to the B. cereus nucleotide sequence coding therefor, to the transformation of microorganisms of the genus E. coli with a plasmid containing this sequence, and to a process for the preparation of the enzyme.
摘要翻译：本发明涉及具有LeuDH活性的酶，其编码的蜡状芽孢杆菌核苷酸序列，用含有该序列的质粒转化大肠杆菌属的微生物，以及制备该酶的方法。

5. 发明授权

US5523223A Ketoester reductase for conversion of keto acid esters to optically active hydroxy acid esters 失效
公开(公告)号：US5523223A
公开(公告)日：1996-06-04
申请号：US284600
申请日：1994-08-11
申请人： Maria-Regina Kula , Jorg Peters
发明人： Maria-Regina Kula , Jorg Peters
IPC分类号： C12N9/02 , C12N9/04 , C12P7/00 , C12P7/62 , C12P41/00 , C12R1/01 , C12R1/38 , C12R1/72 , C12P7/26
CPC分类号： C12N9/0006 , C12P7/62 , Y10S435/921
摘要： A keto ester reductase capable of being used in an NADH-dependent enzymatic reaction for converting .beta., .gamma. and .delta. ketonic acid esters into the corresponding optically active .beta., .gamma. and .delta. hydroxycarboxylic acid esters can be isolated from strains of Candida parapsilosis, Yarrowinia cellobiosa, Rhodococcus erythropolis or Pseudomonas acidovorans, preferably cultivated on a long -chain alkane and/or alkane acid-containing culture medium, approximately in the presence of an inductor. The microorganism is preferrably, Candida parapsilosis DSM 70125. A usable enzyme preparation can be recovered by fractionated PEG-precipitation from the cell raw extract: high specific activities (for example 1855 U/mg) may then be obtained by chromatographic purification. The keto ester reductase is characterized as having a molecular weight of 136 kDa+11 kDa as determined by gel permeation chromatography on Sephadex G-200, a pH optimum for conversion of the keto ester to the hydroxy acid esters between pH 7.8 and 8.0 and for the reverse reaction of converting the acid esters to the keto esters at a pH optimum of 9.5, and a temperature optimum between 36.degree. and 40.degree. C. for conversion of the keto esters to the hydroxy acid esters and from 50.degree. to 56.degree. C. for the reverse reaction of converting the acid esters to the keto esters. Not only (possibly substituted) so-called ketonic esters are accepted, but also number of other oxo-compounds among which diketones, (possibly substituted, in particular halogenated) aliphatic alicyclic and aromatic ketones, as well as ketoacetals and aldehydes. The S-enantiomer-forming reduction is supplemented by the possibility to recover R-enantiomers from racemates by oxidizing the S-enantiomer and separating the oxo-compound.

6. 发明授权

US5238838A L-carnitine amidase produced by a microorganism 失效
标题翻译：由微生物生产的L-酪氨酸酶
公开(公告)号：US5238838A
公开(公告)日：1993-08-24
申请号：US835860
申请日：1992-02-18
申请人： Maria-Regina Kula , Ulrich Joeres
发明人： Maria-Regina Kula , Ulrich Joeres
IPC分类号： C12N1/20 , C12N9/80 , C12P41/00 , C12R1/01
CPC分类号： C12N9/80
摘要： The microorganism DSM 6230 produces an enzyme, L-carnitine amidase. This microorganism and/or the enzyme which it produces can selectively hydrolyze L-carnitine amide and/or the L-component of DL-carnitine amide to L-carnitine.
摘要翻译：微生物DSM 6230产生酶L-肉碱酰胺酶。该微生物和/或其产生的酶可以选择性地将左旋肉碱酰胺和/或左旋肉碱酰胺的L-组分水解成L-肉毒碱。

7. 发明授权

US6001974A Method of separating albumin from serum by ion-exchange chromatography with membrane adsorbers 失效
标题翻译：通过离子交换色谱法用膜吸附器从血清中分离白蛋白的方法
公开(公告)号：US6001974A
公开(公告)日：1999-12-14
申请号：US41082
申请日：1998-03-10
申请人： Wolfgang Demmer , Heinrich Klaus Gebauer , Maria-Regina Kula , Dietmar Nussbaumer , Jorg Thoemmes
发明人： Wolfgang Demmer , Heinrich Klaus Gebauer , Maria-Regina Kula , Dietmar Nussbaumer , Jorg Thoemmes
IPC分类号： C07K1/18 , C07K14/765
CPC分类号： C07K14/765
摘要： A method of separating albumin from serum by ion exchange chromatography with membrane adsorbers characterized by high productivity and yields of high purity albumin. The separation of the albumin is carried out on highly basic anion exchange membranes and on highly acidic cation exchange membranes. The albumin fraction can be eluted from the anion exchange membrane such that it can be fed directly to the cation exchange membrane without any special conditioning and the albumin can be extracted therefrom as an end product.
摘要翻译：通过离子交换色谱法分离白蛋白的方法，其中膜吸附剂的特征在于高生产率和高纯度白蛋白的产率。白蛋白的分离是在高度碱性的阴离子交换膜和高度酸性的阳离子交换膜上进行的。白蛋白部分可以从阴离子交换膜中洗脱出来，使得其可以直接进料至阳离子交换膜而无需任何特殊的调理，并且可以从其中提取白蛋白作为终产物。

8. 发明授权

US5416019A Rhodococcus microorganisms that produce phenylalanine dehydroganase 失效
标题翻译：产生苯丙氨酸脱氢酶的红球菌微生物
公开(公告)号：US5416019A
公开(公告)日：1995-05-16
申请号：US186501
申请日：1994-01-26
申请人： Wolfgang Leuchtenberger , Maria-Regina Kula , Werner Hummel , Horst Schutte
发明人： Wolfgang Leuchtenberger , Maria-Regina Kula , Werner Hummel , Horst Schutte
IPC分类号： C12N1/20 , C12N9/06 , C12P13/04 , C12P13/22 , C12R1/01
CPC分类号： C12N9/0018 , C12N9/0028 , C12P13/04 , C12P13/222 , C12R1/01
摘要： The invention is directed to obtaining phenylalanine-dehydrogenase containing microorganisms by a special selection process comprising obtaining microorganisms, especially strains of Rhodococcus and the phenylalanine-dehydrogenase obtained in them. With the help of this enzyme there are obtained L-.alpha.-aminoacids from the corresponding ketocarboxylic acids by reductive amination.
摘要翻译：本发明涉及通过特殊的选择方法获得含苯丙氨酸脱氢酶的微生物，该方法包括获得微生物，尤其是获得的红球菌菌株和苯丙氨酸脱氢酶。在这种酶的帮助下，通过还原胺化得到相应的酮羧酸的L-α-氨基酸。

9. 发明授权

US5369016A Peptide amidase and the use thereof 失效
标题翻译：肽酰胺酶及其用途
公开(公告)号：US5369016A
公开(公告)日：1994-11-29
申请号：US005819
申请日：1993-01-19
申请人： Doerte Steinke , Maria-Regina Kula , Alexander Schwarz , Christian Wandrey
发明人： Doerte Steinke , Maria-Regina Kula , Alexander Schwarz , Christian Wandrey
IPC分类号： C07K1/107 , C12N9/78 , C12N9/80 , C12P13/04 , C12P21/00 , C12P21/02
CPC分类号： C07K1/107 , C12N9/80
摘要： A peptide amidase isolated from the flavedo of citrus fruits, preferably oranges, which is capable of catalyzing the selective hydrolytic elimination of the free amino group on the C-terminal end of peptide amides but which does not cleave peptide bonds. The enzyme accepts D-amino acid residues in the C-terminal position, although the hydrolysis rate is much slower than with L-amino acid residues. The enzyme is weakly inhibited by serine protease inhibitors; has an optimal pH of 7.5.+-.1.5, an optimum temperature of 30.degree. C. at pH 7.5 and has an isoelectric point of pH 9.5. The peptide amidase is stable at pH 6.0-9.0. The molecular weight of the purified enzyme is 32,000.+-.3000 daltons. A peptide amidase according to the present invention is particularly useful in the production of peptides by continuous enzymatic reaction of N-protected amino acid or peptide alkyl esters with amides of amino acids. In the continuous reaction, the synthesized peptide amide is hydrolyzed by the peptide amidase and separated by anion exchange from the amide of the amino acid which can be recycled.
摘要翻译：从柑橘类水果（优选橙子）中分离的肽酰胺酶，其能够催化肽酰胺的C末端上的游离氨基的选择性水解消除，但不切割肽键。该酶在C-末端位置接受D-氨基酸残基，尽管水解速度比L-氨基酸残基慢得多。酶被丝氨酸蛋白酶抑制剂弱抑制; 最佳pH为7.5 +/- 1.5，最适温度为30℃，pH 7.5，等电点pH值为9.5。肽酰胺酶在pH 6.0-9.0下是稳定的。纯化酶的分子量为32,000 +/- 3000道尔顿。根据本发明的肽酰胺酶特别可用于通过N-保护的氨基酸或肽烷基酯与氨基酸酰胺的连续酶反应来生产肽。在连续反应中，合成的肽酰胺被肽酰胺酶水解，并通过阴离子交换从可再循环的氨基酸的酰胺中分离。

10. 发明授权

US5190875A Peptide amidase and the use thereof 失效
标题翻译：肽类药物及其用途
公开(公告)号：US5190875A
公开(公告)日：1993-03-02
申请号：US694981
申请日：1991-05-06
申请人： Doerte Steinke , Maria-Regina Kula , Alexander Schwarz , Christian Wandrey
发明人： Doerte Steinke , Maria-Regina Kula , Alexander Schwarz , Christian Wandrey
IPC分类号： C07K1/107 , C12N9/78 , C12N9/80 , C12P13/04 , C12P21/00 , C12P21/02
CPC分类号： C07K1/107 , C12N9/80
摘要： A peptide amidase isolated from the flavedo of citrus fruits, preferably oranges, which is capable of catalyzing the selective hydrolytic elimination of the free amino group on the C-terminal end of peptide amides but which does not cleave peptide bonds. The enzyme accepts D-amino acid residues in the C-terminal position, although the hydrolysis rate is much slower than with L-amino acid residues. The enzyme is weakly inhibited by serine protease inhibitors; has an optimal pH of 7.5.+-.1.5, an optimum temperature of 30.degree. C. at pH 7.5 and has an isoelectric point of pH 9.5. The peptide amidase is stable at pH 6.0-9.0. The molecular weight of the purified enzyme is 23,000 +/- 3000 daltons. A peptide amidase according to the present invention is particular useful in the production of peptides by continuous enzymatic reaction of N-protected amino acid or peptide alkyl esters with amides of amino acids. In the continuous reaction, the synthesized peptide amide is hydrolyzed by the peptide amidase and separated by anion exchange from the amide of the amino acid which can be recycled.
摘要翻译：从柑橘类水果（优选橙子）中分离的肽酰胺酶，其能够催化肽酰胺的C末端上的游离氨基的选择性水解消除，但不切割肽键。该酶在C-末端位置接受D-氨基酸残基，尽管水解速度比L-氨基酸残基慢得多。酶被丝氨酸蛋白酶抑制剂弱抑制; 最佳pH为7.5 +/- 1.5，最适温度为30℃，pH 7.5，等电点pH值为9.5。肽酰胺酶在pH 6.0-9.0下是稳定的。纯化酶的分子量为23000 +/- 3000道尔顿。根据本发明的肽酰胺酶特别可用于通过N-保护的氨基酸或肽烷基酯与氨基酸的酰胺的连续酶反应来生产肽。在连续反应中，合成的肽酰胺被肽酰胺酶水解，并通过阴离子交换从可再循环的氨基酸的酰胺中分离。

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