Dsm IP Assets B.v.

Roche Vitamins Inc.

Institut National De La Recherche Agronomique

F. Hoffmann-la Roche Ag

Hans-Peter Hohmann

Adolphus Van Loon

Martin Lehmann

Luis Pasamontes

Michel Tessier

Nigel John Mouncey

Michael Hans

Dietmar Laudert

Uwe Sauer

Jeffrey W Stebbins

Jonathan Alan Medlock

Denis Pompon

Markus Wyss

Heinrich Winfred Schlieker

Laurent Lefort

Harald Pichler

Muriel Merkamm

Guenter Pappenberger

Sybille Ebert

Mae Joanne Aguila

Nicola Zamboni

Regina Leber

Corinna Odar

Barbara Petschacher

Stephane Aymerich

Birgit Ploier

Markus H&uuml;mbelin

Walter Schurter

Stéphane Aymerich

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 novel and inventive process for the production of sorbitol from D-sucrose.

Production of D-sorbitol by hydrogenation

The present invention provides an improved biotechnological production of riboflavin (also referred herein as vitamin B2) through modification in the operon containing the riboflavin biosynthetic genes (rib operon), in particular modifications of/in the leader sequences (rib leader) upstream of the corresponding riboflavin biosynthetic genes (rib operon). Furthermore, the present invention relates to genetically engineered microorganisms carrying said modified sequences, processes to generate said modified sequences/microorganisms and the use thereof for production of riboflavin.

Production of riboflavin

This invention relates to the production of 7-dehydrocholesterol, 25-hydroxy-7-dehydrocholesterol, and 25-hydroxy ergosterol in yeast such as. It also relates to various enzymes catalyzing the reduction of the double bond at position 24 of lanosterol, dimethyl zymosterol, methyl zymosterol, zymosterol, cholesta-7, 24-dienol, or cholesta-5,7,24-trienol; or the hydroxylation at position 25 of ergosterol, 7-dehydrocholesterol, cholesta-8-enol, and cholesta-7-enol. It also relates to various nucleic acids encoding cholesterol C25-hydroxylases and sterol &#x394;24-reductases and their use to produce and hydroxylate 7-dehydrocholesterol or ergosterol. It also relates to the yeast strains so produced, and methods of making these sterols that include the steps of cultivaton a transformed yeast cell, and harvesting the resulting sterol(s).

Production of non-yeast sterols by yeast

The present invention relates to a improved process for the biotechnological production of compounds for which ribose-5-phosphate, ribulose-5-phosphate or xylulose-5-phosphate is biosynthetic precursor like riboflavin (vitamin B), FAD, FMN, pyridoxal phosphate (vitamin B), guanosine, GMP, adenosine, AMP. The invention further pertains to the generation of the organism producing those compounds. It furthermore relates to the generation of mutated transketolases that allow normal growth on glucose but reduced growth on gluconate when introduced into the production strains and to polynucleotides encoding them.

Modified transketolase and use thereof

The present invention is directed to novel genes mediating the carbon catabolite repression (CCR) of gluconeogenic genes. Furthermore, the polypeptides encoded by said genes as well as the use of said genes in a process for the production of a target fermentation product is provided. Processes for generating such microorganisms are also provided by the present invention. The invention is also related to a genetically engineered microorganism and its use for the production of a target fermentation product, wherein the gluconeogenic genes are relieved from CCR within said microorganism.

Method for producing a fermentation product using a genetically modified microorganism with a mutation to relieve carbon catabolite repression

The present invention provides a process for producing a target fermentation product. This process includes providing a fermentation medium containing a recombinantly-produced microorganism that over-produces a fermentation product and contains a mutation which causes auxotrophic growth of the microorganism wherein the auxotrophy within the microorganism does not compromise the ability of the microorganism to produce the fermentation product. The medium is then supplied in excess with all substrates required for production of the fermentation product and in growth limiting amounts with a substrate complementing the auxotrophy. Host cells, vectors, and polynucleotide sequences used in the process are also provided. The polynucleotide sequences of the present invention include sequences derived from the biotin operon ofand in particular the bioFDB gene cassette.

Process for producing a target fermentation product

A process for the preparation of a target fermentation product comprising cultivation of an aerobic microorganism to produce such product, wherein in the microorganism the efficiency of the respiratory chain is increased, transformed and novel nucleotide sequences.

Fermentation process

The present invention provides a process for producing a target fermentation product. This process includes providing a fermentation medium containing a recombinantly-produced microorganism that over-produces a fermentation product and contains a mutation which causes auxotrophic growth of the microorganism wherein the auxotrophy within the microorganism does not compromise the ability of the microorganism to produce the fermentation product. The medium is then supplied in excess with all substrates required for production of the fermentation product and in growth limiting amounts with a substrate complementing the auxotrophy. Host cells, vectors, and polynucleotide sequences used in the process are also provided. The polynucleotide sequences of the present invention include sequences derived from the biotin operon of , and in particular the bioFDB gene cassette.

Polynucleotide portions of the biotin operon from B. subtilis for use in enhanced fermentation

Novel proteins of Flavobacterium sp. R1534 and the DNA sequences which encode these proteins are disclosed which provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoid precursors and carotenoids, especially &bgr;-carotene, lycopene, zeaxanthin and cantaxanthin.

Fermentative carotenoid production

The present invention provides a recombinant bacterium for the over-production of riboflavin. The recombinant bacterium has has been transformed by three or four vectors, two of which each comprise either a DNA sequence coding for the riboflavin synthesizing enzymatic activities of,or a DNA sequence which is substantially homologous and one or more transcription elements and a third and/or fourth vector comprising either a DNA sequence coding for the ribA gene product of,or a DNA sequence which is substantially homologous and optionally a transcription element whereby one or a plurality of copies of each of these vectors has/have been integrated at three or four different sites within the bacterium's chromosome.

Riboflavin production

Novel proteins of Flavobacrerium sp. R1534 and the DNA sequences which encode these proteins are disclosed which provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoid precursors and carotenoids, especially .beta.-carotene, lycopene, zeaxanthin and cantaxanthin. Processes are also provided for preparing zeaxanthin by culturing a transformed host cell containing an expression cassette that includes a polynucleotide having a DNA sequence which encodes the GGPP synthase of Flavobacterium sp. R1534 (crtE), the prephytoene synthase of Flavobacterium sp. R1534 (crtB), the phytoene desaturase of Flavobacterium sp. R1534 (crtI), the lycopene cyclase of Flavobacterium sp. R1534 (crtY), or the .beta.-carotene hydroxylase of Flavobacterium sp. R1534 (crtZ). The polynucleotide is substantially free of other polynucleotides of Flavobacterium sp. R1534. The process further includes isolating the zeaxanthin from such cells or the culture medium.

Novel proteins of Flavobacterium sp. R1534 and the DNA sequences which encode these proteins are disclosed which provide an improved biosynthetic pathway from farnesyl pyrophosphate and isopentyl pyrophosphate to various carotenoid precursors and carotenoids, especially .beta.-carotene, lycopene, zeaxanthin and cantaxanthin.

Growing community of inventors

Loerrach, Germany

Hans-Peter Hohmann