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公开(公告)号：US5840570A
公开(公告)日：1998-11-24
申请号：US345018
申请日：1994-11-23
CPC分类号： C12N9/6481 , A23C19/04 , C12N15/80 , C12N9/62 , C12R1/665
摘要： This invention relates to novel mutant filamentous fungi which are deficient in the gene for the corresponding aspartic proteinase. These organisms are useful production hosts in the production of heterologous polypeptides such as chymosin.
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公开(公告)号：US4935369A
公开(公告)日：1990-06-19
申请号：US26422
申请日：1987-03-16
申请人： Kirk J. Hayenga , Virgil B. Lawlis
发明人： Kirk J. Hayenga , Virgil B. Lawlis
CPC分类号： C12N9/6481 , A23C19/0326 , C07K1/1133 , C12N1/06 , C12N15/70 , C12N9/52 , Y10S435/816
摘要： This invention relates to bovine rennin and prorennin in the form of refractile bodies.
摘要翻译： 本发明涉及以折射体形式的牛肾素和肾上腺素。
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公开(公告)号：US5801034A
公开(公告)日：1998-09-01
申请号：US356042
申请日：1994-12-14
CPC分类号： C12N1/005 , Y10S435/814 , Y10S435/911 , Y10S435/917 , Y10S435/942 , Y10S530/823 , Y10S530/824
摘要： This invention provides a method for killing cells in fermentation mixtures in order to prepare the fermentation mixture for processing to recover or extract a desired product from the fermentation mixture. A preferred method of this invention comprises in either order, adjusting the pH of the fermentation mixture to a value equal to or less than about two pH units below the pK.sub.a of the compatible organic acid using a mineral acid, and adding a sufficient amount of a compatible organic acid and/or organic acid salt to the mixture to effect a substantially complete cell kill. The method of this invention is useful for killing microorganisms such as yeast, bacteria or fungi in any culture or fermentation mixture and is particularly useful in systems where it is desired to kill the cells without lysing them.
摘要翻译： 本发明提供了一种杀死发酵混合物中的细胞的方法，以便制备用于从发酵混合物中回收或提取所需产物的加工的发酵混合物。 本发明的优选方法包括以下任一顺序：使用无机酸将发酵混合物的pH调节至等于或小于相容有机酸的pKa的约两个pH单位的值，并加入足量的 相容的有机酸和/或有机酸盐，以实现基本上完全的细胞杀灭。 本发明的方法可用于在任何培养物或发酵混合物中杀死微生物如酵母，细菌或真菌，并且特别适用于希望在不裂解细胞的情况下杀死细胞的系统。
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公开(公告)号：US5130255A
公开(公告)日：1992-07-14
申请号：US627287
申请日：1990-12-13
CPC分类号： G01N33/68 , Y10T436/12 , Y10T436/200833
摘要： Container systems for stable storage of pharmaceutical compounds are disclosed. The container systems prevent the formation of adducts between pharmaceutical compounds and aldehydes during storage of these compounds in containers sealed with elastomeric stoppers that leach aldehydes. Methods for determining the presence of these adducts are also disclosed.
摘要翻译： 公开了用于药物化合物稳定储存的容器系统。 在将这些化合物储存在用浸出醛的弹性塞子密封的容器中时，容器系统防止在药物化合物和醛之间形成加合物。 还公开了确定这些加合物的存在的方法。
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公开(公告)号：US5041377A
公开(公告)日：1991-08-20
申请号：US611967
申请日：1990-11-13
申请人： Todd Becker , Virgil B. Lawlis, Jr.
发明人： Todd Becker , Virgil B. Lawlis, Jr.
IPC分类号： C12N9/56
CPC分类号： C12N9/54 , Y10S435/803 , Y10S435/814 , Y10S435/816 , Y10S435/832 , Y10S435/839
摘要： Crystalline subtilisin is produced by adding a halide salt, such as sodium chloride or calcium chloride, to a concentrated subtilisin solution (at least about 40 g/1). This process does not produce amorphous subtilisin even at high salt concentrations in the solution. Optionally, subtilisin seed crystals also may be added to the concentrate to speed up the crystallization process.
摘要翻译： 通过向浓缩的枯草杆菌蛋白酶溶液（至少约40g / l）中加入卤化物盐如氯化钠或氯化钙来制备结晶枯草杆菌蛋白酶。 该方法即使在溶液中的高盐浓度下也不会产生无定形枯草杆菌蛋白酶。 任选地，也可以将枯草杆菌蛋白酶晶种加入到浓缩物中以加速结晶过程。
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公开(公告)号：US5578463A
公开(公告)日：1996-11-26
申请号：US284942
申请日：1994-08-02
IPC分类号： C12N1/15 , C12N1/19 , C12N9/34 , C12N9/58 , C12N9/64 , C12N15/62 , C12N15/80 , C12N15/09 , C12N15/56
CPC分类号： C12N9/2428 , C12N15/62 , C12N15/80 , C12N9/58 , C12N9/6481 , C07K2319/00 , C07K2319/02 , C07K2319/50 , C07K2319/61
摘要： Novel vectors are disclosed for expressing and secreting heterologous polypeptides from filamentous fungi. Such vectors are used in novel processes to express and secrete such heterologous polypeptides. The vectors used for transforming a filamentous fungus to express and secrete a heterologous polypeptide include a DNA sequence encoding a heterologous polypeptide and a DNA sequence encoding a signal sequence which is functional in a secretory system in a given filamentous fungus and which is operably linked to the sequence encoding the heterologous polypeptide. Such signal sequences may be the signal sequence normally associated with the heterologous polypeptides or may be derived from other sources. The vector may also contain DNA sequences encoding a promoter sequence which is functionally recognized by the filamentous fungus and which is operably linked to the DNA sequence encoding the signal sequence. Preferably functional polyadenylation sequences are operably linked to the 3' terminus of the DNA sequence encoding the heterologous polypeptides. Each of the above described vectors are used in novel processes to transform a filamentous fungus wherein the DNA sequences encoding the signal sequence and heterologous polypeptide are expressed. The thus synthesized polypeptide is thereafter secreted from the filamentous fungus.
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公开(公告)号：US4935354A
公开(公告)日：1990-06-19
申请号：US927778
申请日：1986-11-05
申请人： Kirk J. Hayenga , Virgil B. Lawlis
发明人： Kirk J. Hayenga , Virgil B. Lawlis
CPC分类号： C12N9/6481 , A23C19/0326 , C07K1/1133 , C12N1/06 , C12N15/70 , C12N9/52
摘要： Rennin for making cheese is obtained from insoluble refractile bodies of a recombinant microbial host cell. The rennin is obtained by rupturing the recombinant host cell, isolating and solubilizing the insoluble refractile bodies, and recovering active rennin. Recombinant techniques involve preparing cDNA corresponding to the coding sequence for calf rennin, introducing into an expression vector and expressing in a host cell. As much as 200 mg rennin per liter of culture may be recovered. Prorennin or preprorennin may be produced and rennin derived therefrom.
摘要翻译： 用于制作奶酪的Rennin由重组微生物宿主细胞的不溶性折射体获得。 通过破坏重组宿主细胞，分离和溶解不溶性折射体，并回收活性肾素获得肾素。 重组技术涉及制备对应于小牛肾素编码序列的cDNA，引入表达载体并在宿主细胞中表达。 每升培养物可以回收多达200毫克的肾素。 可以产生丙氨宁或前肾上腺素，并从中衍生肾素。
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公开(公告)号：US06509171B1
公开(公告)日：2003-01-21
申请号：US09048473
申请日：1998-03-26
IPC分类号： C12P2102
CPC分类号： C12N9/6481 , A23C19/04 , C12N9/62 , C12N15/80 , C12R1/665
摘要： This invention relates to novel mutant filamentous fungi which are deficient in the gene for the corresponding aspartic proteinase. These organisms are useful production hosts in the production of heterologous polypeptides such as chymosin.
摘要翻译： 本发明涉及缺乏相应天冬氨酸蛋白酶基因的新型突变丝状真菌。 这些生物体是生产异源多肽如凝乳酶的有用生产宿主。
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9.发明授权Killing cells without lysis in a method for enzyme recovery from a fermentation broth 失效在从发酵液中回收酶的方法中杀死细胞而不溶解
公开(公告)号：US5378621A
公开(公告)日：1995-01-03
申请号：US57851
申请日：1993-05-07
CPC分类号： C12N1/005 , Y10S435/814 , Y10S435/911 , Y10S435/917 , Y10S435/942 , Y10S530/823 , Y10S530/824
摘要： This invention provides a method for killing fungal cells without lysing in fermentation processes in order to prepare the fermentation mixture for processing to recover or extract an extracellularly expressed enzyme from the fermentation mixture. A preferred method of this invention comprises adjusting the pH of the fermentation mixture to less than 2.79 using a mineral acid, then adding sufficient acetic acid to the mixture to affect a substantially complete cell kill in mixture. A salt of the acetic acid can be used. The organic acid or salt can be added, then the pH adjusted to the desired level. Other organic acids can be used, in which case the pH of the mixture is adjusted to the pK.sub.a of the selected organic acid before the organic acid is added to the mixture. The method of this invention is useful for stopping the growth and killing the cells in any micro-organism, culture or fermentation such as those containing yeast, bacteria or fungi and is particularly useful in systems where it is desired to kill the cells without lysing them.
摘要翻译： 本发明提供了一种杀死真菌细胞而不在发酵过程中裂解的方法，以便制备用于从发酵混合物中回收或提取细胞外表达的酶的发酵混合物。 本发明的优选方法包括使用无机酸将发酵混合物的pH调节至小于2.79，然后向混合物中加入足够的乙酸以影响混合物中基本上完全的细胞杀灭。 可以使用乙酸的盐。 可以加入有机酸或盐，然后将pH调节至所需水平。 可以使用其它有机酸，在这种情况下，在将有机酸加入到混合物中之前，将混合物的pH调节至所选有机酸的pKa。 本发明的方法可用于阻止生长和杀死任何微生物，培养或发酵中的细胞，例如含有酵母，细菌或真菌的细胞，并且特别适用于希望在不裂解细胞的情况下杀死细胞的系统 。
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公开(公告)号：US5364770A
公开(公告)日：1994-11-15
申请号：US413010
申请日：1989-09-25
IPC分类号： C12N1/15 , C12N1/19 , C12N9/34 , C12N9/58 , C12N9/64 , C12N15/62 , C12N15/80 , C12N15/09 , C12N15/56
CPC分类号： C12N9/2428 , C12N15/62 , C12N15/80 , C12N9/58 , C12N9/6481 , C07K2319/00 , C07K2319/02 , C07K2319/50 , C07K2319/61
摘要： Novel vectors are disclosed for expressing and secreting heterologous polypeptides from filamentous fungi. Such vectors are used in novel processes to express and secrete such heterologous polypeptides. The vectors used for transforming a filamentous fungus to express and secrete a heterologous polypeptide include a DNA sequence encoding a heterologous polypeptide and a DNA sequence encoding a signal sequence which is functional in a secretory system in a given filamentous fungus and which is operably linked to the sequence encoding the heterologous polypeptide. Such signal sequences may be the signal sequence normally associated with the heterologous polypeptides or may be derived from other sources. The vector may also contain DNA sequences encoding a promoter sequence which is functionally recognized by the filamentous fungus and which is operably linked to the DNA sequence encoding the signal sequence. Preferably functional polyadenylation sequences are operably linked to the 3' terminus of the DNA sequence encoding the heterologous polypeptides. Each of the above described vectors are used in novel processes to transform a filamentous fungus wherein the DNA sequences encoding the signal sequence and heterologous polypeptide are expressed. The thus synthesized polypeptide is thereafter secreted from the filamentous fungus.
摘要翻译： 公开了用于从丝状真菌表达和分泌异源多肽的新型载体。 这些载体用于表达和分泌这些异源多肽的新方法中。 用于转化丝状真菌以表达和分泌异源多肽的载体包括编码异源多肽的DNA序列和编码在给定丝状真菌中在分泌系统中有功能的信号序列的DNA序列，并且可操作地连接到 编码异源多肽的序列。 这样的信号序列可以是通常与异源多肽相关的信号序列，或者可以来自其他来源。 载体还可以含有编码启动子序列的DNA序列，其由丝状真菌功能识别，并且可操作地连接于编码信号序列的DNA序列。 优选地，功能性多腺苷酸化序列可操作地连接到编码异源多肽的DNA序列的3'末端。 上述载体中的每一种用于转化丝状真菌的新方法，其中表达编码信号序列和异源多肽的DNA序列。 由此合成的多肽此后由丝状真菌分泌。
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