Dsm IP Assets B.v.

Roche Vitamins Inc.

Hoffmann-la Roche Inc.

Dsm Nutritional Products Inc.

Dsm Assets B.v.

Masako Shinjoh

Tatsuo Hoshino

Akira Asakura

Noribumi Tomiyama

Nigel John Mouncey

Setsuko Ojima

Connie Lee

Bastien Chevreux

Taro Miyazaki

Anja Meury

Anne F Mayer

Corina Hauk

Christine Toepfer

Akiko Fujiwara

Alan Berry

Bertus Pieter Koekman

Tatsuya Kiyasu

Dick Schipper

Anne Françoise Mayer

Adrianus W H Vollebregt

Markus G Goese

Bastian Chevreux

Marina Van Gelder

Manuela Dalluege

Marie-Gabrielle Beuzelin-Ollivier

Teruhide Sugisawa

Masaaki Tazoe

Yutaka Setoguchi

Andrea Muffler

Takahide Kon

Akio Fujiwara

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.

Vitamin C production in a microorganism, gluconobacter

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

Method for production of vitamin C

The present invention relates to microorganisms genetically engineered to increase yield and/or efficiency of biomass production from a carbon source, such as e.g. glucose. Processes for generating such microorganisms are also provided by the present invention. The invention also relates to polynucleotide sequences comprising genes that encode proteins that are involved in the bioconversion of a carbon source such as e.g. glucose into biomass. 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. Also included are processes of using the polynucleotides and modified polynucleotide sequences to transform host microorganisms leading to a microorganism with reduced carbon source diversion, i.e. higher yield and/or efficiency of biomass production from a carbon source such as e.g. glucose.

Gene GMS 01

The present invention relates to microorganisms genetically engineered to increase yield and/or efficiency of biomass production from a carbon source, such as e.g. glucose. Also included are processes of using the polynucleotides and modified polynucleotide sequences to transform host microorganisms leading to a microorganism with reduced carbon source diversion, i.e. higher yield and/or efficiency of biomass production from a carbon source such as e.g. glucose.

Process for production of biomass from a carbon source

The present invention discloses an isolated polynucleotide molecule derived from a polynucleotide encoding a polypeptide having L-sorbosone dehydrogenase activity comprising a partial nucleotide sequence of at least 20 consecutive nucleotides of SEQ ID NO:1. The present invention further relates to a process for the production of L-ascorbic acid in high yield, in particular a process using resting cells of a microorganism able to convert given carbon sources into vitamin C. The thus obtained vitamin C may be further processed by purification and/or separation steps.

Microbial production of L-ascorbic acid

Fermentative vitamin C production

Method for production of biomass using a gluconobacter oxydans comprising an inactivated NADP dependent glucose dehydrogenase gene

Gene STS 18

The present invention is directed to a novel cytochrome c oxidase complex, genetic materials useful for the preparation of the said complex, such as recombinant polypeptides involved in cytochrome c oxidase complex, recombinant DNA fragments, expression vectors, recombinant organisms and the like. Those novel cytochrome c oxidase complex and genetic materials may be originated from a microorganism having the identifying characteristics ofDSM 4025. The present invention also provides a method for the preparation of the said novel recombinant cytochrome c oxidase complex and a process for the production of 2-keto-L-gulonic acid (2KGA).

Cytochrome c oxidase enzyme complex

Polypeptides and encoding polynucleotides for microbial production of L-ascorbic acid and associated methods

A process for producing L-ascorbic acid from 2-keto-L-gulonic acid or D-erythorbic acid from 2-keto-D-gluconic acid by contacting 2-keto-L-gulonic acid or 2-keto-D-gluconic acid, respectively, with an enzym having &#x3b1;-amylase activity in solution. The solvent for this reaction can be water, an aqueous alcohol, an organic solvent or a mixture thereof. In each case, the starting material can be in the form of the free acid, the sodium salt, or the calcium salt.

Enzymatic process for the manufacture of L-ascorbic acid and D-erythorbic acid

The present invention is directed to a novel cytochrome c oxidase complex, genetic materials useful for the preparation of the said complex, such as recombinant polypeptides involved in cytochrome c oxidase complex, recombinant DNA fragments, expression vectors, recombinant organisms and the like. Those novel cytochrome c oxidase complex and genetic materials may be originated from a microorganism having the identifying characteristics of Gluconobacter oxydans DSM 4025. The present invention also provides a method for the preparation of the said novel recombinant cytochrome c oxidase complex and a process for the production of 2-keto-L-gulonic acid (2KGA).

A process for producing L-ascorbic acid or D-erythorbic acid, or in each case its sodium, potassium or calcium salt, from 2-keto-L-gulonic acid or 2-keto-D-gluconic acid, or in each case its sodium, potassium or calcium salt, that involves incubating 2-keto-L-gulonic acid or 2-keto-D-gluconic acid, each as the free acid or as its sodium, potassium or calcium salt, and the cells of a thermoacidophilic microorganism at temperatures from about 30&deg; C. to about 100&deg; C., and at a pH from about 1 to about 6, in a solution, to form L-ascorbic acid or D-erythorbic acid or an appropriate salt thereof, and isolating said product from the solution.

Microbial process for producing L-ascorbic acid, D-erythorbic acid, and salts thereof

Growing community of inventors

Kanagawa, Japan

Masako Shinjoh