Dsm IP Assets B.v.

Dsm Fine Chemicals Austria Nfg Gmbh & Co. Kg

Dpx Holdings B.v.

Harald Pichler

Helmut Schwab

Wolfgang Skranc

Gerhard Steinbauer

Marcel G Wubbolts

Peter Pojarliev

Joannes Kierkels

Manuela Hermann

Christoph Zenzmaier

Martin Kietzmann

Micheal Stanek

Hans-Peter Hohmann

Martin Lehmann

Oliver May

Michael Stanek

Otto Martin Lehmann

Rudolf Gijsbertus Marie Luiten

Regina Leber

Amin El-Heliebi

Corinna Odar

Barbara Petschacher

Christine Winkler

Birgit Ploier

Andreas Braun

Mirela Ivancic

Holly Stolterfoht

Marcel Wubbolts

The present invention is related to production of a sterol mix in a modified yeast cell, wherein the amount of zymosterol present in said mix is dramatically reduced or abolished via modification of sterol acyltransferase activity within said yeast. The modified yeast cell can be used for production of vitamin D3 or derivatives and/or metabolites thereof.

Production of sterols in modified yeast

The present invention relates to a recombinantstrain characterized in that the organisms contains a gene coding for a protein having esterase activity. Suchis suitable for the preparation of a protein with esterase activity, whereby the expression takes place without coexpression of GroEL and/or GroES from a plasmid. The expression of glutathione reductase and/or thioredoxin reductase may be abolished by mutation. Furthermore, the ability of the organism to grow under oxygen-rich conditions has been restored by mutation. The expression of the gene of the protein with esterase activity is unaccompanied by the expression of an additional gene encoding a heat shock chaperone protein.

Preparation of an esterase

The invention relates to an isolated polypeptide having esterase activity comprising an amino acid sequence shown in any one of SEQ ID NO's 2, 4, 6, 8, 10, 12 or 14 or a homologue thereof, comprising an amino acid substitution or deletion of one or more amino acids as shown in said SEQ ID NO's and resulting in a mutant polypeptide having an increased concentration of the fraction of the mutant polypeptide being present as an active and soluble protein in cleared lysate of the mutant polypeptide expressed inrelative to the concentration of the fraction of the polypeptide without the mutation being present as an active and soluble protein in cleared lysate of the polypeptide without the one or more deletion or substitution expressed inunder the same conditions. The invention also relates to nucleic acid encoding the polypeptides according to the invention, and the use of the polypeptides.

Pig liver esterases

Polypeptide and recombinant protein having esterase activity which exhibit the amino acid sequence SEQ. ID. No. 1 and the use thereof.

Polypeptide having esterase activity and recombinant esterase and use thereof

Method of resolving stereoselectively racemic 2-alkyl-5-halopent-4-ene-carboxylic esters of the formula (I) 

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Deutschlandsberg, Austria

Harald Pichler