Dsm IP Assets B.v.

Roche Vitamins Inc.

Nigel John Mouncey

Masako Shinjoh

Hans-Peter Hohmann

Bastien Chevreux

Corina Hauk

Anne F Mayer

Anja Meury

Bertus Pieter Koekman

Dick Schipper

Connie Lee

Markus G Goese

Christine Toepfer

Adrianus W H Vollebregt

Marina Van Gelder

Bastian Chevreux

Heinrich Winfred Schlieker

Jeffrey W Stebbins

Marie-Gabrielle Beuzelin-Ollivier

Manuela Dalluege

Martin Lehmann

Markus Wyss

Uwe Sauer

Sybille Ebert

Andrea Muffler

Hans-Peter Hohnmann

Nicola Zamboni

The present invention relates to newly identified microorganisms capable of direct production of L-ascorbic acid (hereinafter also referred to as Vitamin C). The invention also relates to polynucleotide sequences comprising genes that encode proteins which are involved in the synthesis of Vitamin C. The invention also features polynucleotides comprising the full length polynucleotide sequences of the novel genes and fragments thereof, the novel polypeptides encoded by the polynucleotides and fragments thereof, as well as their functional equivalents. The present invention also relates to the use of said polynucleotides and polypeptides as biotechnological tools in the production of Vitamin C from microorganisms, whereby a modification of said polynucleotides and/or encoded polypeptides has a direct or indirect impact on yield, production, and/or efficiency of production of the fermentation product in said microorganism. Also included are methods/processes of using the polynucleotides and modified polynucleotide sequences to transform host microorganisms. The invention also relates to genetically engineered microorganisms and their use for the direct production of Vitamin C.

Fermentative Vitamin C production

Microorganism selected fromor, wherein a gene encoding a polypeptide with the activity of a repressor of L-sorbosone dehydrogenase (SNDH) and L-sorbose dehydrogenase (SDH) is disrupted. The gene has a polynucleotide selected from polynucleotides encoding a polypeptide comprising the amino acid sequence of which is represented by SEQ ID NO:2, polynucleotide comprising the nucleotide sequence according to SEQ ID NO:1, polynucleotides the complementary strand of which hybridizes under stringent conditions to a polynucleotide as defined above and which encodes a polypeptide with the activity of a repressor of SNDH and SDH. The microorganism produces at least 10% more Vitamin C and/or 2-KGA from L-sorbose compared to a microorganism wherein the repressor is intact.

Gene SMS 27

The present invention relates to newly identified genes that encode proteins that are involved in the synthesis of L-ascorbic acid (hereinafter also referred to as Vitamin C). The invention also features polynucleotides comprising the full-length polynucleotide sequences of the novel genes and fragments thereof, the novel polypeptides encoded by the polynucleotides and fragments thereof, as well as their functional equivalents. The present invention also relates to the use of the polynucleotides and polypeptides as biotechnological tools in the production of Vitamin C from microorganisms, whereby a modification of the polynucleotides and/or encoded polypeptides has a direct or indirect impact on yield, production, and/or efficiency of production of the fermentation product in the microorganism. Also included are methods/processes of using the polynucleotides and modified polynucleotide sequences to transform host microorganisms. The invention also relates to genetically engineered microorganisms and their use for the direct production of Vitamin C.

Method for production of vitamin C

Fermentative vitamin C production

Gene STS 18

The present invention relates to modified GTP cyclohydrolase II enzymes that display increased specific activity, and to polynucleotides encoding them. The invention further pertains to vectors comprising these polynucleotides and host cells containing such vectors. The invention provides a method for producing the modified enzyme and a method for producing riboflavin, a riboflavin precursor, FMN, FAD, or a derivative thereof.

Mutant GTP cyclohydrolase II enzymes

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

Growing community of inventors

Binningen, Switzerland

Nigel John Mouncey