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    Secretory signal selection vectors for extracellular protein synthesis in Bacilli
    3.
    发明授权
    Secretory signal selection vectors for extracellular protein synthesis in Bacilli 失效
    分泌细胞外蛋白质合成信号选择载体

    公开(公告)号：US5212070A

    公开(公告)日：1993-05-18

    申请号：US627028

    申请日：1990-12-13

    申请人： Hilde E. Smith ,  Jan H. Van Ee ,  Ben P. H. Peeters ,  Sierd Bron ,  Gerard Venema

    发明人： Hilde E. Smith ,  Jan H. Van Ee ,  Ben P. H. Peeters ,  Sierd Bron ,  Gerard Venema

    IPC分类号： C12N1/21 ,  C12N15/62 ,  C12N15/75

    CPC分类号： C12N15/75 ,  C12N15/625 ,  C07K2319/036 ,  C07K2319/61

    摘要： Novel vectors are provided for identifying secretory signal sequences from DNA fragments of unicellular microorganisms. The plasmids comprise a multiple cloning site with restriction sites in reading frame with a structural gene which permits rapid screening of the secreted expression product. Optionally, the vectors may include a promoter region upstream from the multiple cloning site. The invention is exemplified with Bacillus. Specific secretory signal sequences have been isolated with those vectors, allowing for efficient secretion into the supernatant, and not just to the periplasmic space to provide proteins in economically high yields. Secretory sequences are provided superior to other previously known sequences.

    摘要翻译： 提供了用于从单细胞微生物的DNA片段鉴定分泌信号序列的新载体。 质粒包含具有限制性位点的多克隆位点，其具有能够快速筛选分泌的表达产物的结构基因的阅读框。 任选地，载体可以包括来自多克隆位点上游的启动子区。 本发明以芽孢杆菌为例。 已经用这些载体分离了特异性分泌信号序列，允许有效分泌到上清液中，而不仅仅是在周质空间中以经济上高的产率提供蛋白质。 分泌序列被提供优于其他先前已知的序列。

    Attaching substances to micro-organisms
    4.
    发明授权
    Attaching substances to micro-organisms 有权
    将物质附着于微生物

    公开(公告)号：US07169383B1

    公开(公告)日：2007-01-30

    申请号：US09554354

    申请日：1998-11-12

    申请人： Girbe Buist ,  Cornelis Johannes Leenhouts ,  Gerard Venema ,  Jan Kok

    发明人： Girbe Buist ,  Cornelis Johannes Leenhouts ,  Gerard Venema ,  Jan Kok

    IPC分类号： A01N63/00 ,  A01N65/00

    CPC分类号： C12N9/2414 ,  A01N63/02 ,  A61K38/00 ,  A61K39/00 ,  C07K14/195 ,  C07K14/315 ,  C07K14/37 ,  C07K2319/00 ,  C12N9/2408 ,  C12N9/2462 ,  C12N9/86

    摘要： The invention relates to surface display of proteins on micro-organisms via the targeting and anchoring of heterologous proteins to the outer surface of cells such as yeast, fungi, mammalian and plant cells, and bacteria. The invention provides a proteinaceous substance comprising a reactive group and at least one attaching peptide which comprises a stretch of amino acids having a sequence corresponding to at least a part of the consensus amino acid sequence listed in FIG. 10 and comprises a method for attaching a proteinaceous substance to the cell wall of a micro-organism comprising the use of said attaching peptide.

    摘要翻译： 本发明涉及通过将异源蛋白质靶向和锚定到诸如酵母，真菌，哺乳动物和植物细胞以及细菌的细胞的外表面上的微生物上蛋白质的表面显示。 本发明提供了包含反应性基团和至少一种附着肽的蛋白质物质，其包含一段氨基酸，其氨基酸序列对应于图1中列出的共有氨基酸序列的至少一部分。 并且包括将蛋白质物质附着到包含使用所述附着肽的微生物的细胞壁的方法。

    Complex inducible promoter system derivable from a phage of a lactic acid bacterium (LAB), and its use in a LAB for production of a desired protein
    7.
    发明授权
    Complex inducible promoter system derivable from a phage of a lactic acid bacterium (LAB), and its use in a LAB for production of a desired protein 失效
    可从乳酸菌（LAB）的噬菌体衍生的复合诱导型启动子系统及其在LAB中用于产生所需蛋白质的用途

    公开(公告)号：US6143525A

    公开(公告)日：2000-11-07

    申请号：US737226

    申请日：1997-04-03

    申请人： Arjan Nauta ,  Gerard Venema ,  Jan Kok ,  Adrianus Marinus Ledeboer

    发明人： Arjan Nauta ,  Gerard Venema ,  Jan Kok ,  Adrianus Marinus Ledeboer

    IPC分类号： C12N15/09 ,  C12N1/21 ,  C12N9/64 ,  C12N15/74 ,  C12P21/02 ,  C12R1/01 ,  C12P21/04

    CPC分类号： C12N15/746

    摘要： The invention provides a complex inducible promoter system from a phage of a lactic acid bacterium, especially one having the DNA sequence of SEQ. ID. No: 3 given in FIG. 2, or a DNA sequence essentially corresponding to those sequences, and a modification of (an essential part of) such promoter system in which the mitomycin C induction system is replaced by a good-grade system, e.g. a temperature-initiated induction system or a salt-initiated induction system. Also is provided a recombinant vector and a transformed lactic acid bacterium comprising (an essential part of) such promoter system. Further a process for producing a desired protein by such transformed bacterium is provided, comprising expressing a gene encoding said desired protein or a precursor thereof under control of such promoter system or an essential part thereof. Preferably, the transformed lactic acid bacterium is made food-grade due to using food-grade DNA sequences and/or removing non-food-grade DNA sequences. When required, the desired protein can be secreted by the lactic acid bacterium if a DNA sequence fused to the gene encoding the desired protein is present which effects secretion of the desired protein or its precursor. The process can be used in a fermentation process, in which the desired protein causes lysis of the bacterial cells so that the contents of the cells can be released, or in which the desired protein is an enzyme involved in flavour formation, or in which the desired protein has a function in a cheese production process, such as chymosin or a precursor thereof, or an enzyme involved in flavour formation.

    摘要翻译： PCT No.PCT / NL95 / 00172 Sec。 371日期1997年4月3日 102（e）日期1997年4月3日PCT提交1995年5月12日PCT公布。 公开号WO95 / 31563 日期：1995年11月23日本发明提供了来自乳酸菌噬菌体的复合诱导型启动子系统，特别是具有SEQ ID NO： ID。 图3中给出。 2或基本上对应于这些序列的DNA序列，以及其中丝裂霉素C诱导系统被优良系统替代的（启动子系统的（其主要部分）的修饰）。 温度启动的诱导系统或盐引发的诱导系统。 还提供了重组载体和包含这种启动子系统的（必要部分）的转化的乳酸菌。 此外，提供了通过这种转化细菌产生所需蛋白质的方法，包括在该启动子系统或其主要部分的控制下表达编码所述期望蛋白质或其前体的基因。 优选地，由于使用食品级DNA序列和/或去除非食品级DNA序列，所述转化的乳酸菌被制成食品级。 当需要时，如果存在与编码所需蛋白质的基因融合的DNA序列，则所需的蛋白质可以由乳酸菌分泌，其影响所需蛋白质或其前体的分泌。 该方法可用于发酵过程中，其中期望的蛋白质引起细菌细胞的裂解，使得细胞的内容物可以释放，或其中所需的蛋白质是涉及风味形成的酶，或其中 所需蛋白质在奶酪生产过程中具有功能，例如凝乳酶或其前体，或涉及风味形成的酶。

    Salt-inducible promoter derivable from a lactic acid bacterium, and its use in a lactic acid bacterium for production of a desired protein
    8.
    发明授权
    Salt-inducible promoter derivable from a lactic acid bacterium, and its use in a lactic acid bacterium for production of a desired protein 失效
    衍生自乳酸菌的盐诱导型促进剂，以及其在乳酸菌中用于产生所需蛋白质的用途

    公开(公告)号：US6140078A

    公开(公告)日：2000-10-31

    申请号：US68195

    申请日：1998-07-29

    申请人： Jan W. Sanders ,  Jan Kok ,  Gerard Venema ,  Adrianus M Ledeboer

    发明人： Jan W. Sanders ,  Jan Kok ,  Gerard Venema ,  Adrianus M Ledeboer

    IPC分类号： C12N15/09 ,  C07K14/335 ,  C12N1/21 ,  C12N15/74 ,  C12R1/01 ,  C12P21/06

    CPC分类号： C12N15/746 ,  C07K14/335

    摘要： The invention provides a salt-inducible promoter present in SEQ ID NO: 10 and derivable from a lactic acid bacterium in isolation from the coding sequence normally controlled by said promoter in a wild-type lactic acid bacterium, with modifications and important parts thereof. Also provided are a recombinant vector and a transformed lactic acid bacterium comprising such promoter, and the production of a desired protein by such transformed bacterium, whereby the gene encoding said desired protein or a precursor thereof is expressed under control of such promoter. The desired protein can be secreted by the bacterium due to the presence of a signal sequence. The action of the salt-inducible promoter is enhanced at a pH of about 4-4.5 and/or by the presence of glutamic acid. Such process can be used in a fermentation process, in which the desired protein is a lytic protein causing lysis of the cells and release of the cell content. Or the desired protein can be an enzyme involved in flavour formation, e.g. in producing dressings, spreads, sausages and sour dough, or a protein functioning in a cheese production process, such as chymosin or a precursor thereof, or an enzyme involved in cheese flavour formation.

    摘要翻译： PCT No.PCT / EP97 / 04755 Sec。 371日期1998年7月29日第 102（e）日期1998年7月29日PCT 1997年8月20日PCT PCT。 公开号WO98 / 10080 PCT 日期1998年3月12日本发明提供了存在于SEQ ID NO：10中的衍生自乳酸菌的盐诱导型启动子，其与野生型乳酸菌通常由所述启动子控制的编码序列分离， 其重要部分。 还提供了包含这种启动子的重组载体和转化的乳酸菌，并且通过这种转化细菌产生所需的蛋白质，由此编码所述所需蛋白质或其前体的基因在这种启动子的控制下表达。 由于信号序列的存在，所需的蛋白质可被细菌分泌。 在约4-4.5的pH和/或谷氨酸的存在下，盐诱导型启动子的作用增强。 这种方法可用于发酵过程中，其中所需蛋白质是引起细胞裂解和释放细胞内容物的裂解蛋白。 或者所需的蛋白质可以是参与风味形成的酶，例如 在制作敷料，酱油，香肠和酸面团中，或在酪乳生产过程中起作用的蛋白质，例如凝乳酶或其前体，或涉及奶酪香味形成的酶。

    Process for the lysis of a culture of lactic acid bacteria by means of a lysin, and uses of the resulting lysed culture
    9.
    发明授权
    Process for the lysis of a culture of lactic acid bacteria by means of a lysin, and uses of the resulting lysed culture 失效
    通过溶素裂解乳酸菌的培养的方法，以及所得裂解培养物的用途

    公开(公告)号：US5955258A

    公开(公告)日：1999-09-21

    申请号：US737716

    申请日：1997-04-22

    申请人： Girbe Buist ,  Gerard Venema ,  Jan Kok ,  Adrianus Marinus Ledeboer

    发明人： Girbe Buist ,  Gerard Venema ,  Jan Kok ,  Adrianus Marinus Ledeboer

    IPC分类号： A23C19/032 ,  A61K8/00 ,  A61K8/60 ,  A61K8/64 ,  A61K8/66 ,  A61K8/99 ,  A61Q19/00 ,  A61Q19/10 ,  C07H21/04 ,  C12N1/21 ,  C12N9/14 ,  C12N15/09 ,  C12N15/55 ,  C12N15/74 ,  C12P21/06 ,  C12R1/01 ,  C12R1/19 ,  C12Q1/00 ,  A01N63/00 ,  C12N1/00

    CPC分类号： C12N9/14 ,  A23C19/0323 ,  C12N15/746 ,  C12P21/06

    摘要： The invention provides a process for the lysis of a culture of lactic acid bacteria, or a product containing such culture e.g. cheese, by means of a lysin through the in situ production of a homologous autolysin, or a heterologous autolysin obtainable from Gram-positive bacteria esp. from lactic acid bacteria. The gene encoding said autolysin is controlled by a promoter, preferably regulated by food-grade ingredients or parameters, to achieve an enhanced lysis after induction resulting in an enhanced production of total autolysin compared with the natural production lever of the homologous autolysin during fermentation or shortly thereafter. Other uses of the invention include preparing a mixture of peptides which are modified by peptidases freed after the lysis, using the autolysin as a bactericidal agent against spoiling bacteria or pathogenic bacteria for improving the shelf life of a product containing the lysed culture.

    摘要翻译： PCT No.PCT / NL95 / 00170 Sec。 371日期1997年04月22日 102（e）日期1997年4月22日PCT提交1995年5月12日PCT公布。 WO95 / 31561 PCT出版物 日期：1995年11月23日本发明提供了乳酸菌培养物的裂解方法，或含有这种培养物的产物。 干酪，通过原位产生同源自溶素的溶素，或可从革兰氏阳性菌esp获得的异源自溶素。 来自乳酸菌。 编码所述自溶素的基因由优选由食品级成分或参数调节的启动子控制，以在诱导后实现增强的裂解，从而与发酵期间的同源自溶素的天然生产杠杆相比较，或者短期内 之后 本发明的其它用途包括制备由裂解后释放的肽酶修饰的肽的混合物，使用自溶素作为防止破坏细菌或病原菌的杀菌剂，以改善含有裂解培养物的产品的保质期。

    Attaching substances to microorganisms
    10.
    发明申请
    Attaching substances to microorganisms 有权
    将物质附着于微生物

    公开(公告)号：US20070104724A1

    公开(公告)日：2007-05-10

    申请号：US11590738

    申请日：2006-10-30

    申请人： Girbe Buist ,  Cornelis Leenhouts ,  Gerard Venema ,  Jan Kok

    发明人： Girbe Buist ,  Cornelis Leenhouts ,  Gerard Venema ,  Jan Kok

    IPC分类号： A61K39/395 ,  C07K16/46 ,  C07K14/195

    CPC分类号： C12N9/2414 ,  A01N63/02 ,  A61K38/00 ,  A61K39/00 ,  C07K14/195 ,  C07K14/315 ,  C07K14/37 ,  C07K2319/00 ,  C12N9/2408 ,  C12N9/2462 ,  C12N9/86

    摘要： The invention relates to surface display of proteins on microorganisms via the targeting and anchoring of heterologous proteins to the outer surface of cells such as yeast, fungi, mammalian, plant cells, and bacteria. The invention provides a proteinaceous substance comprising a reactive group and at least one attaching peptide including a stretch of amino acids having a sequence corresponding to at least a part of the consensus amino acid sequence listed in FIG. 10 and further includes a method for attaching a proteinaceous substance to the cell wall of a microorganism comprising the use of the attaching peptide.

    摘要翻译： 本发明涉及通过将异源蛋白质靶向和锚定到细胞如酵母，真菌，哺乳动物，植物细胞和细菌的外表面上来微生物上蛋白质的表面显示。 本发明提供一种蛋白质物质，其包含反应性基团和至少一种附着肽，其包括具有对应于图1中列出的共有氨基酸序列的至少一部分的序列的一段氨基酸。 并且还包括将蛋白质物质附着到包含使用附着肽的微生物的细胞壁中的方法。