Yamasa Corporation

Hokkaido University

Shizuoka University

Yamasa Shoyu Kabushiki Kaisha

Shizuoka Prefectural Universities Corporation

Toshitada Noguchi

Tomoki Hamamoto

Kazuya Ishige

Kiyoshi Okuyama

Toshikazu Shiba

Kenji Takenouchi

Takashi Suzuki

Shin-Ichiro Nishimura

Yuichiro Midorikawa

Yusuke Ohba

Ilpal Jwa

Taiichi Usui

Takeomi Murata

Kuniaki Nagaoka

Noriko Nagahori

Takashi Kawakami

Hideo Takahashi

The invention provides a method for producing 3'-phosphoadenosine 5&#x2032;-phosphosulfate (PAPS), the method including subjecting ATP to sulfation and phosphorylation by use of adenosine 5&#x2032;-triphosphate sulfurylase (ATPS) and adenosine 5&#x2032;-phosphosulfate kinase (APSK), wherein an adenosine 5&#x2032;-triphosphate (ATP) supply/regeneration system including adenosine 5&#x2032;-monophosphate (AMP), polyphosphate, polyphosphate-driven nucleoside 5&#x2032;-diphosphate kinase (PNDK), and polyphosphate:AMP phosphotransferase (PAP), or an adenosine 5&#x2032;-triphosphate (ATP) supply/regeneration system including adenosine 5&#x2032;-monophosphate (AMP), polyphosphate, polyphosphate-driven nucleoside 5&#x2032;-diphosphate kinase (PNDK), and adenylate kinase (ADK) is employed instead of ATP.

Method of enzymatically synthesizing 3'-phosphoadenosine-5&#x2032;-phosphosulfate

DNA fragment encoding a polyphosphate-driven nucleoside 5'-diphosphate kinase polypeptide

A method by which high-purity CMP-N-acetylneuraminic acid (HPLC purity, 95% or higher), which has been difficult to obtain with any technique other than chromatography, can be easily obtained in satisfactory yield by a simple operation without the need of chromatography. The process, which is for producing high-purity CMP-N-acetylneuraminic acid (CMP-NeuAc), is characterized by conducting a suitable combination of the following steps (1) to (4). Step 1: a step in which divalent cations are added to a solution containing CMP-NeuAc to thereby precipitate the phosphoric acid, pyrophosphoric acid, and nucleotide which coexist; Step 2: a step in which a phosphatase is added to a solution containing CMP-NeuAc to thereby convert the coexistent nucleotide into nucleoside; Step 3: a step in which an organic solvent is added to precipitate the CMP-NeuAc; and Step 4: a step in which the CMP-NeuAc precipitated is recovered.

Process for producing CMP-N-acetylneuraminic acid

A polymer containing an N-linked sialo-glycan wherein a sialo-glycan is condensed to a &#x3b3;-polyglutamic acid using a chemical compound having an amino group on one end and a carboxyl group on another end and represented by the structural formula (I). Formula (I) (In the formula, Z means a hydroxy group or a residue represented by the formula (II), and n represents an integer of 10 or more, with the proviso that any one or more of the Z's is represented by the formula in (II).) Formula (II) (In the formula, X means a hydroxy group or an acetylamino group, Yand Ymean a hydroxyl group or an N-acetylneuraminic acid residue, L means a hydrocarbon, an m represents 0 or an integer of 1 or 2, with the proviso that Yand Yare not the same.) 

Synthetic N-linked sialo-glycan-containing polymer and method for producing the same

The present invention is directed to, for example, an oligosaccharide having at an end thereof a 4-position halogenated galactose residue represented by formula (I): (wherein X represents a halogen atom, and R represents a monosaccharide, an oligosaccharide, or a carrier), a transferase inhibitor containing the oligosaccharide, and a method for inhibiting sugar chain elongation reaction in the presence of glycosyltransferase, the method including employing the inhibitor. The invention also provides a method for producing a 4-position halogenated galactose sugar nucleotide represented by formula (II): (wherein each of Rto Rrepresents a hydroxyl group, an acetyl group, a halogen atom, or a hydrogen atom; X represents a halogen atom; and M represents a hydrogen ion or a metal ion), wherein the method employs bacterium-derived galactokinase and bacterium-derived hexose-1-phosphate uridylyltransferase. The invention is also directed to a sugar chain containing 4-position halogenated galactose envisaged to be employed as drugs and other materials, and to applications of the compound.

Sugar chain containing 4-position halogenated galactose and application thereof

The present invention is directed to a process for producing CMP-N-acetylneuraminic acid (CMP-NeuAc), comprising adding to the culturedcells which has been transformed with both the DNA encoding N-acetylglucosamine-6-phosphate 2-epimerase (GlcNAc-6P 2-epimerase) and the DNA encoding N-acetylneuraminic acid synthase (NeuAc synthase) and exhibit activities of N-acetylglucosamine-6-phosphate 2-epimerase and N-acetylneuraminic acid synthase, a phosphate buffer containing baker's yeast cells, CMP, N-acetylglucosamine (GlcNAc), magnesium, xylene, glucose, and CMP-N-acetylneuraminic acid synthase (CMP-NeuAc synthase) to provide a reaction mixture, and allowing the reaction to proceed and produce CMP-N-acetylneuraminic acid (CMP-NeuAc), and wherein the process does not require adding ATP.

The present invention provides a method for enzymatically producing uridine 5'-diphospho-N-acetylgalactosamine (UDP-GalNAc) (which is an important substrate for oligosaccharide synthesis) from uridine 5&#x2032;-triphosphate (UTP) and N-acetylgalactosamine 1-phosphate (GalNAc 1-P), the method including using, as an enzyme, uridine 5&#x2032;-diphospho-N-acetylglucosamine pyrophosphorylase (UDP-GlcNAc pyrophosphorylase) derived from a microorganism (exclusive of a pathogenic microorganism). The GalNAc 1-P employed can be prepared from N-acetylgalactosamine and a phosphate donor in a reaction system by use of N-acetylgalactosamine kinase. According to the present invention, uridine 5&#x2032;-diphospho-N-acetylgalactosamine can be efficiently produced by use of a relatively inexpensive substrate.

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The present invention provides a stable salt of 3'-phosphoadenosine 5&#x2032;-phosphosulfate (PAPS) and a production method therefor. The present invention is directed to a stable salt of PAPS (amine salt), which is formed between PAPS and an amine compound, and to a method for producing a stable salt of PAPS, which includes adding an amine compound to an aqueous PAPS solution in an amount by mole equal to or greater than that of PAPS, and lyophilizing the resultant solution. The present invention has first realized production of a solid-form PAPS salt having considerably improved stability through a very simple technique. Since the thus-produced amine salt of PAPS is very stable, the salt can be stored or employed without taking much care about decomposition thereof at ambient temperature.

Stable salt of 3'-phosphoadenosine 5&#x2032;-phosphosulfate

This invention relates to a novel polyphosphate: AMP phosphotransferase (PAP), a gene coding this PAP, and their use. The PAP has the following properties: (A) action: catalyzing of the following two reactions:NMP+PolyP&#x2192;NDP+PolyPdNMP+POlyP&#x2192;dNDP+PolyP(wherein NMP represents nucleoside monophosphate, NDP represents nucleoside diphosphate, dNMP represents deoxynucleoside monophosphate, dNDP represents deoxynucleoside diphosphate, n represents degree of polymerization of the polyphosphate which is an integer of up to 100); (B) substrate specificity: specific to AMP, GMP, IMP, dAMP, and dGMP, also acting with CMP, UMP, dCMP, and TMP; (C) molecular weight: about 55 to 56 Kd (kilodalton); and (D) specific activity: at least 70 units per 1 mg of enzyme protein.

Polyphosphate:AMP phosphotransferase

The present invention relates to a new use of uridine diphosphate glucose 4-epimerase (also called uridine diphosphate galactose 4-epimerase), and a method of converting uridine diphosphate N-acetylglucosamine (UDP-GlcNAc) to uridine diphosphate N-acetylgalactosamine (UDP-GalNAc) by using the said enzyme. The process for producing UDP-GalNAc by using the uridine diphosphate glucose 4-epimerase and the UDP-GalNAc supply system according to the present invention are practical and efficient, and greatly beneficial to the industries.

Use of uridine diphosphate glucose 4-epimerase

A process for producing uridine diphosphate-N-acetylglucosamine (UDPAG) from uridylic acid (UMP) and N-acetylglucosamine by use of microorganism cells, characterized by adding N-acetylglucosamine kinase thereto. According to the present invention, UDPAG can be efficiently produced even when N-acetylglucosamine is used as a substrate.

Process for producing uridine diphosphate-n-acetylglucosamine

Process for preparing sugar nucleotide

Process for producing nucleoside 5'-triphosphates and application of the

A process of producing a foreign protein is provided which comprises transforming E. coli with a plasmid carrying a fused DNA having a DNA fragment encoding a foreign protein located downstream of a transcription initiation signal and translation initiation signal both derived from an appropriate T4 phage gene, infecting the transformant with a T4 phase denB and/or alc mutant at a low multiplicity of infection, and then culturing the infected transformant. The desired foreign protein can be produced in an extremely high yield, as compared to the prior art technique.

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Choshi, Japan

Toshitada Noguchi