Mitsubishi Petrochemical Company Limited

Mitsubishi Chemical Corporation

Research Association for Utilization of Light Oil

Petroleum Energy Center

Hideaki Yukawa

Masato Terasawa

Terukazu Nara

Yasurou Kurusu

Makoto Goto

Hisashi Yamagata

Yoshihiro Takayama

Mitsunobu Shimazu

Makiko Fukushima

Yukie Satoo

Keiko Kohama

Thomas Joel Zupancic

Kazuhisa Hatakeyama

Yasukazu Uchida

Izuru Tokumaru

Nobutake Fugono

Kazuoki Osumi

Fuzio Endo

Youichi Niimura

Michio Kozaki

Miki Ikuta

Makiko Kukushima

Masato Terasausa

Miki Kobayashi

Terukatsu Nara

Fumaric acid is produced by reacting a culture of a microorganism which produces maleate isomerase that exhibits a maximum activity at not less than 50.degree. C. or a treated product thereof with maleic acid in an aqueous solution, and isomerizing maleic acid to produce fumaric acid. L-aspartic acid is produced by reacting both of a culture of a microorganism which produces maleate isomerase or a treated product thereof and a culture of a microorganism which produces aspartase or a treated product thereof with maleic acid and ammonia in an aqueous solution, producing L-aspartic acid from maleic acid and ammonia by enzyme reactions, and recovering L-aspartic acid from the reaction mixture.

Method of producing fumaric acid

The present invention relates to a process for producing L-tryptophan in a single-stage reaction, comprising carrying out an L-tryptophan producing reaction with glycine, formaldehyde and indole as raw materials in an aqueous solution in the presence of microbial cells having serine transhydroxymethylase or a treated products thereof and microbial cells having tryptophan synthase or tryptophanase, or a treated products thereof; and collecting produced L-tryptophan from the reaction solution.

Process for the preparation of L-tryptophan

An oligonucleotide comprising a specific region and one or two non-specific regions which are bound to at least one of the two termini of the specific region, wherein the specific region has a specific base sequence which is substantially complementary to a target sequence of a sample nucleic acid with which the oligonucleotide is to be hybridized, the non-specific region is composed of at least one nucleotide having other base capable of forming a base pair with each of bases constituting a common nucleic acid, or its oligomer, is used as a probe for the hybridization.

Oligonucleotide and method for analyzing base sequence of nucleic acid

Coryneform bacteria promoter DNA fragments are disclosed. The promoter function of the promoter DNA fragments is controllable by replacing at least one of the culture-medium ingredients with at least one other ingredient. The sizes and nucleotide sequences of such promoter DNA fragments are also disclosed.

Promoter DNA fragment from coryneform bacteria

Coryneform bacteria promoter DNA fragments are disclosed having greater promoter strength in Coryneform bacteria cells than the tac promoter obtained by fusing Escherichia coli trp promoter and lac promoter. The promoter function of some of the promoter DNA fragments is controllable by replacing at least one of the culture-medium ingredients with at least one other ingredient. The sizes and nucleotide sequences of such promoter DNA fragments are also disclosed.

Fumaric acid is produced by reacting maleic acid in an aqueous solution with a microorganism which has maleate isomerase activity or with a preparation from the microorganism having the maleate isomerase activity, and producing fumaric acid in a reaction solution by enzymatic isomerization of maleic acid carried out under the condition that the dissolved oxygen concentration in the reaction solution is substantially maintained at 4 ppm or less, for example, by sealing the reaction solution with one or more gases selected from N.sub.2, Ar, and He.

Method for producing fumaric acid

The present invention relates to a DNA fragment which contains a gene responsible for the function of autonomous replication of plasmid in a Coryneform bacterium, said gene being obtained from plasmid pBY503 held in Brevibacterium stationis, and in which at least one point mutation capable of increasing the copy number of plasmid exists on said gene region. By using a Coryneform transformed with the vector constructed using said DNA fragment and an industrially useful gene such as aspartase gene or tryptophan synthase gene, production of the useful product occurs with higher efficiency than conventional methods.

Coryneform bacteria DNA fragment containing a gene responsible for the

There is disclosed a process for preparing indigo which comprises bringing an indigo-producing bacteria belonging to genus Acinetobacter or treated products thereof into contact with an aqueous solution containing at least indole to have indigo formed and accumulated in the aqueous solution; and collecting indigo from the aqueous solution.

Process for preparing indigo

A process for producing an NADH oxidase which comprises culturing alkalophilic, facultative anaerobic NADH oxidase-producing bacteria having the optimum growth pH in an alkaline region in medium and collecting said NADH oxidase from the culture; and a plasmid used for the said process.

Process for producing NADH oxidase

A process for cultivating a microorganism transformed with a recombinant plasmid at least containing (a) a DNA fragment containing a promoter and a regulator gene tnaC located downstream of the promoter in a tryptophanase operon (tna), and (b) a DNA fragment containing a desired structural gene which can be expressed by the promoter, which comprises cultivating the transformed microorganism in a culture medium while adding glucose as a carbon source continuously or intermittently so that the concentration of glucose is maintained within the range of 0.01 to 0.3%, and thereby allowing the desired structural gene to be expressed in the microorganism.

Cultivation of transformed microorganisms

A method of selectively inhibiting pyruvic acid decomposition activity in microorganism cells containing tryptophanase or a treated product thereof, which comprises heat-treating said cells or the treated product thereof in the presence of an ammonium ion.

Method of treating microorganism cells containing tryptophanase or

A DNA fragment derived from plasmid pBY503 obtained from Brevibacterium stationis IF012144, said DNA fragment containing a gene which encodes for the function of maintaining a plasmid, capable of replicating and proliferating at least in a Coryneform bacteria of the genus Brevibacterium, stably in said bacteria; and a vector DNA capable of replicating and proliferating in a Coryneform bacteria into which the above DNA fragment is introduced.

DNA fragment containing gene which encodes the function of stabilizing

A method for producing L-threonine, which comprises subjecting at least L- or DL-aspartic acid or a salt thereof to enzymatic reaction according to the reaction system not accompanied with growth of microorganism cells in an aqueous solution in the presence of a microorganism and collecting L-threonine formed, wherein the microorganism is a biotin-requiring microorganism for the growth belonging to coryneform bacterium; a plasmid comprising a DNA fragment containing at least a gene encoding biosynthesis of threonine which can be expressed within a biotin-requiring microorganism cell for the growth belonging to coryneform bacterium and a DNA fragment containing a gene encoding autonomous replication within coryneform bacterium cell; and a biotin-requiring microorganism for the growth belonging to coryneform bacterium which has been transformed with the plasmid described above, both of which are employed in the present method. According to the present invention, L-threonine can be produced with good yield, and further since production management becomes extremely easy without requiring cumbersome operation such as sterilization of the medium, etc. as in the fermentation method, L-threonine can be produced inexpensively in industry.

Method for producing L-threonine, and plasmid and microorganism employed

Microorganisms of the genus Pseudomonas containing aspartate beta-decarboxylase at a high level are produced by culturing said microorganism in a medium supplemented with a chelating agent. L-Alanine can be efficiently produced from L-aspartic acid said microorganisms or the treated product thereof.

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Inashiki-gun, Japan

Hideaki Yukawa