摘要:
The present invention relates to improved variants of soluble pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenases (s-GDH), to genes encoding mutated s-GDH, to mutant proteins of s-GDH with improved substrate specificity for glucose, and to different applications of these s-GDH variants, particularly for determining concentrations of sugar, especially of glucose in a sample.
摘要:
A simple and efficient method for random in vitro mutagenesis and recombination of polynucleotide sequences based on walk-through with chain-elongating molecules and chain terminating molecules e.g. dNTPs/ddNTPs followed by reassembly and amplification is described. The utility of this method for improving protein structure and/or function is demonstrated by creating novel sarcosine oxidase variants and by recombining human placental alkaline phosphates and calf intestinal alkaline phosphatase for better mutants with improved properties.
摘要:
The present invention relates to a mutant protein of PQQ-dependent s-GDH characterized in that in at least one of the positions 122 and 124 the amino acid lysine is present, wherein these positions correspond to the amino acid positions known from the A. calcoaceticus s-GDH wild-type sequence (SEQ ID NO: 2), it also discloses genes encoding such mutant s-GDH, and different applications of these sGDH mutants, particularly for determining the concentration of glucose in a sample.
摘要:
The invention relates to recombinant proteinase K and to a method for producing recombinant proteinase K, which is characterised by the following steps: a) transformation of a host cell containing a recombinant nucleic acid that codes for the zymogenic precursor of the proteinase K, b) cultivation of the host cell in such a way that the zymogenic precursor of proteinase K occurs in the form of inclusion bodies in said host cell, c) isolation of the inclusion bodies and re-naturing under conditions, from which the protease part of the zymogenic precursor emerges in its natural conformation, d) activation and purification of the re-natured proteinase K.
摘要:
The invention relates to recombinant proteinase K and to a method for producing recombinant proteinase K, which is characterised by the following steps: a) transformation of a host cell containing a recombinant nucleic acid that codes for the zymogenic precursor of the proteinase K, b) cultivation of the host cell in such a way that the zymogenic precursor of proteinase K occurs in the form of inclusion bodies in said host cell, c) isolation of the inclusion bodies and re-naturing under conditions, from which the protease part of the zymogenic precursor emerges in its natural conformation, d) activation and purification of the re-natured proteinase K.
摘要:
The present invention discloses a mutant of PQQ-dependent soluble glucose dehydrogenase (s-GDH; EC 1.1.5.2) with improved specificity for glucose as compared to maltose, having a substitution of threonine at position 348 by either glycine, alanine or serine, wherein said mutant additionally comprises, at least one mutation for improving the stability of the mutant and one or more mutation(s) for improving the affinity of the mutant to glucose, and/or one or more mutation(s) for further improving the specificity of the mutant for glucose as compared to maltose, and wherein position 348 correspond to the amino acid positions known from the A. calcoaceticus s-GDH wild-type sequence. Also disclosed are genes encoding such mutant s-GDH, and different applications of these s-GDH mutants, particularly for determining the concentration of glucose in a sample.
摘要:
The present invention relates to improved variants of soluble pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenases (s-GDH), to genes encoding mutated s-GDH, to mutant proteins of s-GDH with improved substrate specificity for glucose, and to different applications of these s-GDH variants, particularly for determining concentrations of sugar, especially of glucose in a sample.