公开(公告)号：US06518054B1

公开(公告)日：2003-02-11

申请号：US09381982

申请日：1999-12-07

申请人： Lambertus Van den Burg ,  Oene Robert Veltman ,  Gerard Venema

发明人： Lambertus Van den Burg ,  Oene Robert Veltman ,  Gerard Venema

IPC分类号： C12N900

CPC分类号： A23J3/34 ,  C11D3/386 ,  C11D3/38663 ,  C12N9/54 ,  C12P21/06 ,  C14C1/065

摘要： The present invention provides genes encoding variants of metallo-endopeptidases that have been engineered to be resistant to prolonged boiling while having maintained their enzymatic performance at much lower temperatures. In addition, thermal stability of the metallo-endopeptidases is highly dependent on calcium at concentrations in the mM range. The invention further provides active metallo-endopeptidases variants whose stability depending on calcium concentration can be changed so as to provide metallo-endopeptidases that are calcium dependent or independent. The invention also provides genes that encode boiling-resistant metallo-endopetidases whose stability depending on calcium concentration can be changed. The invention also provides vectors and cells comprising these genes and proteases produced through these genes, vectors and/or cells. In particular variants with the above described properties are provided of thermolysin-like proteases such as produced by Bacillus stearothermophilus (TLP-ste) and Bacillus thermoproteolyticus (thermolysine). Boiling-resistant and calcium independent or dependent metallo-endopeptidases can be applied in several industrial processes, for instance in the preparation of the artificial sweetener aspartame, but also in the leather industry, in breweries and in the production of protein hydrolysates.

摘要翻译： 本发明提供编码金属内肽酶变体的基因，其已被工程化以对延长的煮沸有抵抗力，同时保持其在较低温度下的酶性能。 此外，金属内肽酶的热稳定性高度依赖于在mM范围内的浓度下的钙。 本发明还提供活性金属内肽酶变体，其可以改变其依赖于钙浓度的稳定性，以提供钙依赖性或独立性的金属内肽酶。 本发明还提供编码耐沸腾金属内肽酶的基因，其可以改变其依赖于钙浓度的稳定性。 本发明还提供了通过这些基因，载体和/或细胞产生的包含这些基因和蛋白酶的载体和细胞。 特别是具有上述性质的变体提供了嗜热脂肪芽孢杆菌（Bacillus stearothermophilus）（TLP-ste）和热嗜热芽孢杆菌（Thermolycolyticus）产生的嗜热菌蛋白酶样蛋白酶。 抗沸腾和钙依赖性或依赖性金属内肽酶可以应用于几个工业过程中，例如制备人造甜味剂阿斯巴甜，还可用于皮革工业，酿酒厂和生产蛋白质水解产物。

公开(公告)号：US5683909A

公开(公告)日：1997-11-04

申请号：US135133

申请日：1993-10-12

申请人： Jan Kok ,  Jan Maat ,  Josephus Mauritius van der Vossen ,  Gerard Venema

发明人： Jan Kok ,  Jan Maat ,  Josephus Mauritius van der Vossen ,  Gerard Venema

IPC分类号： C12N15/74 ,  C12N15/75 ,  C12N15/69 ,  C12N15/70

CPC分类号： C12N15/746 ,  C12N15/75

摘要： The claimed invention is drawn to a recombinant plasmid which can replicate in Bacillus subtilis, Escherichia coli, and lactic acid Streptococcus bacteria comprising the replication of origin from Streptococcus cremoris plasmid pWV01 as its origin of replication, in addition to coding marker genes and genes of interest which code for improved fermenting properties.

摘要翻译： 所要求保护的发明涉及一种重组质粒，除了编码标记基因和感兴趣的基因之外，还可以在枯草芽孢杆菌，大肠杆菌和乳酸链球菌中复制，其包含起始于来自链球菌链球菌质粒pWV01的复制物作为其复制起点 该代码用于改善发酵性能。