Abstract: A method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a desired replacement sequence is disclosed.

Abstract: The present invention provides a method for the fermentative production of compactin, lovastatin, pravastatin or simvastatin comprising culturing a host, preferably a filamentous fungus, comprising the polynucleotide of the lovE transcription regulator gene from Aspergillus terreus. Furthermore, the invention provides a host for the production of above mentioned statines comprising the polynucleotide of the lovE transcription regulator gene from Aspergillus terreus.

Abstract: The present invention relates to polynucleotide sequences which enable a polynucleotide control sequence, such as a promoter, to direct expression in a wide range of industrially relevant species, both prokaryotes and eukaryotes.

Abstract: The present invention discloses a polypeptide selected from the group consisting of: a polypeptide having an amino acid sequence according to SEQ ID NO 3, a polypeptide having an amino acid sequence according to SEQ ID NO 6, a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 3 and a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 6, the polypeptide displaying acetamidase activity and providing a reverse selection on fluoroacetamide with an efficiency of at least 95%, preferably at least 96%, more preferably at least 97%, more preferably at least 98%, more preferably at least 99%, most preferably 100%. The gene encoding the polypeptide of the invention is used as an efficient bi-directional selection marker in the construction of selection marker free strains, in particular for processes for the production of a compound of interest.

Abstract: The present invention relates to a method for preparing an adipate ester or thioester. The invention further relates to a method for preparing adipic acid from said ester or thioester. Further the invention provides a number of methods for preparing an intermediate for said ester or thioester. Further the invention relates to a method for preparing 6-amino caproic acid (6-ACA), a method for preparing 5-formyl valeric acid (5-FVA), and a method for preparing caprolactam. Further, the invention relates to a host cell for use in a method according to the invention.

Abstract: The present invention relates to a polypeptide with HMG-CoA reductase activity, to its polynucleotide congener and to a method for the production of a statin comprising over expression of said polypeptide.

Abstract: The present invention provides a polypeptide having an amino acid sequence according to SEQ ID NO 3, SEQ ID NO 6 or SEQ ID NO 43-59. The present invention also provides a polynucleotide comprising a DNA sequence encoding these polypeptides and a method for isolating polynucleotides encoding polypeptides capable of improving the compactin into pravastatin conversion. Furthermore, the present invention provides a method for producing pravastatin and a pharmaceutical composition comprising pravastatin.

Abstract: The present invention provides genetically engineered microbial strains, in particular genetically engineered yeast strains, that produce at least 0.5 mg per g CDW of a sphingoid base according to Formula I or a salt or ester thereof. The present invention provides a method to obtain genetically engineered microbial strains producing at least 0.5 mg per g CDW of a sphingoid base according to Formula I or a salt or ester thereof.

Abstract: The present invention provides a microorganism containing a compactin biosynthesis gene and a gene for conversion of compactin into pravastatin. In a preferred example, said compactin biosynthesis gene is mIcA and/or mIcB and/or mIcC and/or mIcD and/or mIcE and/or mIcF and/or mIcG and/or mIcH and/or mIcR and said gene for conversion of compactin into pravastatin is a hydroxylase gene. Furthermore, the present invention provides a method for producing a compound of interest such as a statin. In a preferred example said statin is pravastatin.

Abstract: The present invention relates to a transformed eukaryotic cell comprising one or more nucleotide sequence(s) encoding acetyl-CoA acetyltransferase, 3-hydroxybutyryl-CoA dehydrogenase, 3-hydroxybutyryl-CoA dehydratase, butyryl-CoA dehydrogenase, alcohol dehydrogenase or acetaldehyde dehydrogenase and/or NAD(P)H-dependent butanol dehydrogenase, whereby the nucleotide sequence(s) upon transformation of the cell confer(s) the cell the ability to produce butanol. The invention also relates to a process for the production of butanol.

Abstract: The present invention provides a method for the transfection of filamentous fungal cells, comprising providing a multitude of containers, filling into each container an amount of polymer needed for the transfection, filling the cells to be transfected as well as an aqueous solution of transfection reagent into each of the containers, incubating the resulting mixture, removing the transfection reagent from the incubated mixture; and selecting the cells which have been transformed, characterized in that the total volume of the incubating mixture is less than 1 ml per container. Furthermore, the present invention provides the use of transformed filamentous fungal cells for the production of proteins or metabolites.

Abstract: The present invention discloses a polypeptide selected from the group consisting of: a polypeptide having an amino acid sequence according to SEQ ID NO 3, a polypeptide having an amino acid sequence according to SEQ ID NO 6, a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 3 and a polypeptide having an amino acid that is substantially homologous to the sequence of SEQ ID NO 6, the polypeptide displaying acetamidase activity and providing a reverse selection on fluoroacetamide with an efficiency of at least 95%, preferably at least 96%, more preferably at least 97%, more preferably at least 98%, more preferably at least 99%, most preferably 100%. The gene encoding the polypeptide of the invention is used as an efficient bi-directional selection marker in the construction of selection marker free strains, in particular for processes for the production of a compound of interest.

Abstract: The present invention provides a recombinant Penicillium chrysogenum strain characterized in that a gene selected from the list consisting of penDE, pcbAB and pcbC is inactivated and the use of such a strain for the preparation of a compound of interest.

Abstract: The present invention provides a fermentative process for the synthesis of simvastatin by providing a host capable of incorporating the 2,2-dimethylbutyrate side chain into simvastatin, i.e. by customizing a polyketide synthase gene optimized for synthesis and/or incorporation of 2,2-dimethylbutyrate; optionally feeding said host with the appropriate substrate for 2,2-dimethylbutyrate synthesis; fermenting said host to obtain simvastatin or analogues or derivatives thereof, i.e. by producing simvastatin on an industrial scale by a fed-batch process.

Abstract: We disclose a method to construct eukaryotic cells having a target sequence in a chromosomal DNA sequence replaced by a desired replacement sequence, comprising: modifying a parent eukaryotic cell with a preference for NHR to provide a eukaryotic cell having an increased HR/NHR ratio as compared to the parent cell, providing a DNA molecule comprising a first DNA fragment comprising a desired replacement sequence flanked at its 5? and 3? sides by DNA sequences substantially homologous to sequences of the chromosomal DNA flanking the target sequence and a second DNA fragment comprising an expression cassette comprising a gene encoding a selection marker operably linked to regulatory sequences functional in the eukaryotic cell, transforming the modified eukaryotic cells with the DNA molecule, selecting transformed progeny cells having the DNA molecule inserted into the chromosome, deselecting transformed progeny cells having the DNA molecule inserted into the chromosome via NHR by expression of the selection mar

Abstract: The present invention discloses a method to construct fungal cells having a target sequence in a chromosomal DNA sequence replaced by a desired replacement sequence, comprising: providing a DNA molecule comprising a first DNA fragment comprising a desired replacement sequence flanked at its 5? and 3? sides by DNA sequences substantially homologous to sequences of the chromosomal DNA flanking the target sequence and a second DNA fragment comprising an expression cassette comprising a gene encoding diphtheria toxin A and regulatory sequences functional in the fungal cell operably linked thereto; transforming the fungal cells with the resulting DNA molecule; growing the cells to obtain transformed progeny cells having the DNA molecule inserted into the chromosome, wherein cells in which the DNA molecule is inserted in the chromosome via a non-homologous recombination event are selectively killed by expression of diphtheria toxin A; and obtaining cells wherein the target sequence in the chromosomal DNA sequence i

Abstract: The present invention provides microbial strains, in particular yeast strains, that produce at least 0.1 mg per g biomass dry weight of a sphingoid base. The present invention further provides a method to obtain sphingoid base-producing microbial strains comprising incubating a population of microbial cells in the presence of a suitable concentration of a toxin, selecting cells that are resistant against said toxin, and isolating cells out of the toxin-resistant cell population that produce at least 0.1 mg per g biomass dry weight of the sphingoid base of Formula I. Optionally, the method further comprises subjecting a population of toxin-resistant microbial cells that produce at least 0.1 mg per g biomass dry weight of the sphingoid base of Formula I to DNA-mediated transformation with a polynucleotide encoding an enzyme of the sphingolipid metabolic pathway. The present invention further provides a polypeptide having dihydroceramide desaturase activity obtainable form Pichia ciferrii.

Abstract: The present invention relates to a method for increasing the efficiency of targeted integration of a polynucleotide to a pre-determined site into the genome of a filamentous fungal cell with a preference for NHR, wherein said polynucleotide has a region of homology with said pre-determined site, comprising steering an integration pathway towards HR. The present invention also relates to a mutant filamentous fungus originating from a parent cell, said mutant having an HR pathway with elevated efficiency and/or an NHR pathway with a lowered efficiency and/or a NHR/HR ratio with decreased efficiency as compared to said HR and/or NHR efficiency and/or NHR/HR ratio of said parent cell under the same conditions.

Abstract: The present invention describes the transformation of a microorganism that does not naturally produce a ?-lactam compound with polynucleotides involved in the biosynthesis of ?-lactam compounds and the use of such transformed microorganisms in the production of ?-lactam compounds or in the identification of genes or factors involved in the synthesis of a ?-lactam compound.