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CLONING, EXPRESSION AND USE OF A PHOSPHOLIPASE FROM ASPERGILLUS FUMIGATUS

机译:烟曲霉中磷脂酶的克隆,表达及用途

摘要

The invention relates to a DNA sequence, which codes for a polypeptide having phospholipase activity essentially without lipase activity, characterized in that the DNA sequence is selected from a) DNA sequences that comprise a nucleotide sequence according to SEQ ID NO: 1, b) DNA sequences that comprise the coding sequence according to SEQ ID NO: 1, c) DNA sequences that code for the protein sequence according to SEQ ID NO: 2, d) DNA sequences that are coded for by the plasmid pPL3940-Topo2.5 with the restriction map according to FIG. 7, which is deposited under accession number DSM 22741, e) DNA sequences that hybridize under stringent conditions with one of the DNA sequences according to a), b), c) or d), f) DNA sequences that are related to the DNA sequences according to a), b), c), d) or e) due to the degeneration of the genetic code, and g) complementary strands to the sequences according to a) to f), wherein the DNA sequence is preferably derived from Aspergillus, and more preferably from Aspergillus fumigatus, and a polypeptide having phospholipase activity essentially without lipase activity selected from a) a polypeptide which is coded for by the coding part of a DNA sequence as defined above, b) a polypeptide having the sequence according to SEQ ID NO: 2 or a sequence derived therefrom, which may be obtained by substitution, addition, deletion of one or more amino acid(s), c) a polypeptide having a sequence that has at least 83% identity with the amino acids 1 to 299 of SEQ ID NO: 2, d) a polypeptide which is coded for by a nucleic acid sequence which hybridizes under stringent conditions with (i) nucleotides 55 to 1106 of SEQ ID NO: 1, (ii) the cDNA sequence contained in nucleotides 55 to 1106 of SEQ ID NO: 1, (iii) a partial sequence of (i) or (ii) composed of at least 100 nucleotides, or (iv) a complementary strand of (i), (ii) or (iii), e) a variant of the polypeptide having SEQ ID NO: 2, comprising a substitution, deletion and/or insertion of one or more amino acid(s), f) allelic variants to amino acid sequences a) to e).
机译:本发明涉及一种DNA序列,其编码具有基本上没有脂肪酶活性的具有磷脂酶活性的多肽,其特征在于该DNA序列选自:a)包含根据SEQ ID NO:1的核苷酸序列的DNA序列。序列,其包含根据SEQ ID NO:1的编码序列,c)编码根据SEQ ID NO:2的蛋白质序列的DNA序列,d)由质粒pPL3940-Topo2.5编码的DNA序列,具有根据图2的限制图。参见图7,其以登录号DSM 22741保藏,e)在严格条件下与根据a),b),c)或d)的DNA序列之一杂交的DNA序列,f)与该DNA相关的DNA序列由于遗传密码的退化而导致的根据a),b),c),d)或e)的序列,以及g)与根据a)至f)的序列互补的链,其中DNA序列优选衍生自具有曲霉菌,更优选来自烟曲霉的多肽,以及具有磷脂酶活性的多肽,该多肽基本上不具有脂肪酶活性,该多肽选自a)通过上述DNA序列的编码部分编码的多肽,b)具有根据SEQ ID NO:2或由其衍生的序列,其可以通过取代,添加,缺失一个或多个氨基酸获得,c)具有与氨基酸1具有至少83%同一性的序列的多肽到SEQ ID NO:2的299至d)的多肽,其中通过在严格条件下与(i)SEQ ID NO:1的核苷酸55-1106杂交的核酸序列编码(ii)SEQ ID NO:1的核苷酸55-1106所包含的cDNA序列(i)或(ii)的至少100个核苷酸组成的部分序列,或(iv)(i),(ii)或(iii)的互补链,e)具有SEQ ID NO: 2,包括一个,多个氨基酸,f)等位基因变体对氨基酸序列a)至e)的取代,缺失和/或插入。

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