Genes/genetic elements associated with mating impairment in Trichoderma reesei QM6a and its derivatives and process for their identification

    公开(公告)号:US09920384B2

    公开(公告)日:2018-03-20

    申请号:US14758238

    申请日:2013-12-23

    申请人: AB Enzymes GmbH

    摘要: The invention relates to a process for identifying gene(s)/genetic element(s) associated with mating impairment in strains of Trichoderma reesei QM6a or strains derived thereof comprising the steps of a) providing a first strain being a Trichoderma reesei QM6a strain having a MAT1-2 locus or a strain derived thereof, b) sexually crossing said strain with a second strain being a mating competent strain of a Trichoderma reesei (Hypocrea jecorina) strain having a complementary locus, i.e. the MAT1-1 locus, c) repeatedly back-crossing the MAT1-1 progenies from the crossing of b) or the back-crossing thereof with the first strain of a), until a strain is obtained that is substantially identical to the first Trichoderma reesei QM6a strain or a strain derived thereof, but carries the MAT1-1 locus and is mating competent for crossing with Trichoderma reesei QM6a or any of its MAT1-2 progeny, d) selecting the progeny from step c) that is mating competent for crossing with a Trichoderma reesei (Hypocrea jecorina) having a MAT1-2 locus, and e) identifying the gene(s)/genetic element(s) associated with mating impairment by comparing the genome of the progenies selected in step d) with the genome sequences of the first strain of a) whereby said gene(s)/genetic element(s) may be fully or partially missing or existing in a mutated form or in a form having deletions or insertions in the first strain thus being a gene or a genetic element directly or indirectly associated with mating impairment in strains of Trichoderma reesei QM6a or a strain derived thereof as well as to a process for correcting the mating impairment of a Trichoderma reesei QM6a strain or a strain derived thereof having a MAT1-1 locus and that is not competent to mate with a Trichoderma reesei QM6a strain having a MAT1-2 locus or a strain derived thereof, wherein one or more mutated or fully or partially missing gene(s) and/or genetic element(s) identified as above is/are replaced by or complemented with the corresponding functional gene(s) and/or genetic element(s). Moreover, the invention relates to the use of a thus obtained fungal strain of the genus Trichoderma in industrial breeding and production of a product of interest. Moreover, the invention relates to the genes associated with mating impairment of Trichoderma reesei QM6a and strains derived therefrom and to genes essential for mating of Trichoderma reesei QM6a and strains derived therefrom.

    Cloning, expression and use of acid phospholipases

    公开(公告)号:US09322003B2

    公开(公告)日:2016-04-26

    申请号:US14692230

    申请日:2015-04-21

    申请人: AB Enzymes GMBH

    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).

    CLONING, EXPRESSION AND USE OF ACID PHOSPHOLIPASES

    公开(公告)号:US20150299680A1

    公开(公告)日:2015-10-22

    申请号:US14692230

    申请日:2015-04-21

    申请人: AB Enzymes GMBH

    IPC分类号: C12N9/18 A23D9/02 A23D9/00

    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).

    GENES/GENETIC ELEMENTS ASSOCIATED WITH MATING IMPAIRMENT IN TRICHODERMA REESEI QM6A AND ITS DERIVATIVES AND PROCESS FOR THEIR IDENTIFICATION

    公开(公告)号:US20150361510A1

    公开(公告)日:2015-12-17

    申请号:US14758238

    申请日:2013-12-23

    申请人: AB ENZYMES GMBH

    摘要: The invention relates to a process for identifying gene(s)/genetic element(s) associated with mating impairment in strains of Trichoderma reesei QM6a or strains derived thereof comprising the steps of a) providing a first strain being a Trichoderma reesei QM6a strain having a MAT1-2 locus or a strain derived thereof, b) sexually crossing said strain with a second strain being a mating competent strain of a Trichoderma reesei (Hypocrea jecorina) strain having a complementary locus, i.e. the MAT1-1 locus, c) repeatedly back-crossing the MAT1-1 progenies from the crossing of b) or the back-crossing thereof with the first strain of a), until a strain is obtained that is substantially identical to the first Trichoderma reesei QM6a strain or a strain derived thereof, but carries the MAT1-1 locus and is mating competent for crossing with Trichoderma reesei QM6a or any of its MAT1-2 progeny, d) selecting the progeny from step c) that is mating competent for crossing with a Trichoderma reesei (Hypocrea jecorina) having a MAT1-2 locus, and e) identifying the gene(s)/genetic element(s) associated with mating impairment by comparing the genome of the progenies selected in step d) with the genome sequences of the first strain of a) whereby said gene(s)/genetic element(s) may be fully or partially missing or existing in a mutated form or in a form having deletions or insertions in the first strain thus being a gene or a genetic element directly or indirectly associated with mating impairment in strains of Trichoderma reesei QM6a or a strain derived thereof as well as to a process for correcting the mating impairment of a Trichoderma reesei QM6a strain or a strain derived thereof having a MAT1-1 locus and that is not competent to mate with a Trichoderma reesei QM6a strain having a MAT1-2 locus or a strain derived thereof, wherein one or more mutated or fully or partially missing gene(s) and/or genetic element(s) identified as above is/are replaced by or complemented with the corresponding functional gene(s) and/or genetic element(s). Moreover, the invention relates to the use of a thus obtained fungal strain of the genus Trichoderma in industrial breeding and production of a product of interest. Moreover, the invention relates to the genes associated with mating impairment of Trichoderma reesei QM6a and strains derived therefrom and to genes essential for mating of Trichoderma reesei QM6a and strains derived therefrom.