Abstract: Lipolytic enzymes are chemically modified by covalently linking one or more (particularly 1-3) hydrophobic groups to the enzyme molecule. The chemical modification improves the performance of the lipolytic enzyme, e.g., in baking or in detergents.

Abstract: The inventors have developed a method using protein engineering to produce lipolytic enzymes having a relatively high activity for one ester bond in an amphiphilic substrate with two lipophilic groups) and a relatively low activity for the ester bond in an amphiphilic substrate with one lipophilic group, e.g. a relatively high phospholipase activity and a relatively low lysophospholipase activity.

Abstract: Lipolytic enzymes are chemically modified by covalently linking one or more (particularly 1–3) hydrophobic groups to the enzyme molecule. The chemical modification improves the performance of the lipolytic enzyme, e.g., in baking or in detergents.

Abstract: Certain variants of Lipolase (wild-type Humicolalanuginosa lipase) have a particularly good first-wash performance in a detergent solution. The variants should comprise one or more substitutions with positive amino acids near the N-terminal in the three-dimensional structure. The variants should further comprise a peptide addition at the C-terminal and/or should meet certain limitations on electrically charged amino acids at positions 90-101 and 210.

Abstract: Methods for screening a polynucleotide library for a polypeptide with a property of interest in a filamentous fungal host cell, in a manner which allows quick and easy subsequent characterization of the polypeptide, using an expression cloning vector comprising at least a polynucleotide encoding a selectable marker in which the translation initiation start site of the marker-encoding sequence comprises a crippled consensus Kozak sequence, a fungal replication initiation sequence, and a promoter with a cloning-site into which the library is cloned, and a transcription terminator.

Abstract: The substrate specificity of a lipolytic enzyme can be modified by making alterations to the amino acid sequence in a defined region of the lipolytic enzyme, so as to increase the level of a desired activity or to decrease the level of an undesired activity. Thus, the inventors have developed lipolytic enzyme variants with a modified amino acid sequence with a substrate specificity which can be tailored for specific uses.

Abstract: A method for making polynucleotide libraries that comprise new recombined polynucleotides by using mismatch repair enzymes or proteins to shuffle at least two variants of the same polynucleotide or at least two homologous polynucleotides through a Recombinatorial Chain Reaction, RCR.

Abstract: Novel lipolytic enzymes are disclosed which are capable of removing substantial amounts of lard from a lard stained swatch in a one cycle wash. Preferred lipolytic enzymes are variants of the Humicola lanuginosa lipase which may be prepared by recombinant DNA techniques. The enzymes are advantageously used in detergent compositions.

Abstract: A method of constructing and screening a library of polynucleotide sequences of interest in filamentous fungal cells by use of an episomal replicating AMA1-based plasmid vector, thus achieving a high frequency of transformation and a stable and standard uniformly high level of gene expression.

Abstract: The inventors have isolated novel genes with a high homology to the T. lanuginosus lipase gene and are thus well suited for use in gene shuffling. Accordingly, the invention provides a method of generating genetic diversity into lipolytic enzymes by family shuffling of two or more homologous genes which encode lipolytic enzymes. The DNA shuffling technique is used to create a library of shuffled genes, and this is expressed in a suitable expression system and the expressed proteins are screened for lipolytic enzyme activity. The expressed proteins may further be screened to identify lipolytic enzymes with improved properties. The invention also provides a polynucleotide comprising a nucleotide sequence encoding a lipolytic enzyme and a lipolytic enzyme (a polypeptide with lipolytic enzyme activity).

Abstract: Attaching a peptide extension to the C-terminal amino acid of a lipase re-duces the tendency to form odor. This may lead to lipase variants with a reduced odor generation when washing textile soiled with fat which includes relatively short-chain fatty acyl groups (e.g. up to C8) such as dairy stains containing butter fat or tropical oils such as coconut oil or palm kernel oil.

Abstract: The present invention relates to a method of heterologous production of a taxane or a related compound by cloning a DNA sequence comprising a taxane synthesis pathway, making a DNA construct wherein said DNA sequence is under control of regulatory elements, introducing said DNA construct into a host cell, growing said host cell under conditions conductive to the production of the taxane in question, and recovering the taxane in question from the culture medium. The invention also relates to the DNA sequence comprising the taxane synthesis pathway, an expression vector comprising the taxane synthesis pathway, and a host cell comprising the expression vector comprising the taxane synthesis pathway being capable of heterologous expression of taxane.

Abstract: The invention relates to a variant of a parent fungal glucoamylase, which exhibits altered properties, in particular improved thermal stability and/or increased specific activity.

Abstract: The present invention is directed to phospholipase from Fusarium solani (MUCL38667) and analogues thereof, nucleic acid constructs, recombinant expression vectors, and recombinant host comprising the nucleic acid sequences encoding the phospholipase from Fusarium solani (MUCL38667) and analogues thereof, methods of making same, and method of using the phosopholipases of the present invention in baking methods, baking compositions and detergent compositions.

Abstract: The present invention relates to a modified enzyme with lipolytic activity, a lipolytic enzyme capable of removing a substantial amount of fatty matter during a one cycle wash, a DNA sequence encoding said enzymes, a vector comprising said DNA sequence, a host cell harbouring said DNA sequence or said vector, and a process for producing said enzymes with lipolytic activity.

Abstract: The present invention relates to lipase variants having at least 90% identity to the wild-type lipase derived from Humicola lanuginosa strain DSM 4109 and having a certain distribution of electrically charged amino acids. The present invention also relates to detergents comprising such lipases.

Abstract: A method for in vitro construction of a library of recombined homologous polynucleotides from a number of different starting DNA templates and primers by induced template shifts during an polynucleotide synthesis is described, whereby A. extended primers are synthesized by a) denaturing the DNA templates b) annealing primers to the templates, c) extending the said primers by use of a polymerase, d) stop the synthesis, and e) separate the extended primers from the templates, B. a template shift is induced by a) isolating the extended primers from the templates and repeating steps A.b) to A.e) using the extended primers as both primers and templates, or b) repeating steps A.b) to A.e), C. this process is terminated after an appropriate number of cycles of process steps A. and B.a), A. and B.b), or combinations thereof. Optionally the polynucleotides are amplified in a standard PCR reaction with specific primers to selectively amplify homologous polynucleotides of interest.

Abstract: The invention relates to a variant of a parent fungal glucoamylase, which exhibits improved thermal stability and/or increased specific activity using saccharide substrates.

Abstract: A process for making DNA libraries in filamentous fungal cells using a novel cloned gene involved in the mismatch repair system of filamentous fungal cells.

Abstract: A method for making polynucleotide libraries that comprise new recombined polynucleotides by using mismatch repair enzymes or proteins to shuffle at least two variants of the same polynucleotide or at least two homologous polynucleotides through a Recombinatorial Chain Reaction, RCR.