Yamasa Corporation

Kazuya Ishige

Toshitada Noguchi

Kenji Takenouchi

Hisaki Tanaka

Kaori Tanabe

Ko Yoshida

Tomoki Hamamoto

Yuichiro Midorikawa

Kiyoshi Okuyama

Marie Matsunaga

Takashi Kawakami

Misaki Takamatsu

A lyophilized product of cyclic-di-AMP requires special production equipment and is thus not suitable for large-scale production. Crystals of cyclic-di-AMP free acid are unstable under severe conditions at 105&#xb0; C. Then, the present invention addresses the problem of providing a cyclic-di-AMP crystal that can be easily acquired in a large amount and is very stable under the severe conditions at 105&#xb0; C. Crystals of c-di-AMP sodium salt according to the present invention are extremely stable even under the severe conditions at 105&#xb0; C. Further, the crystals of c-di-AMP sodium salt according to the present invention can be prepared in a large amount by a simple process including adjusting a c-di-AMP aqueous solution at pH 5.2-12.0 and then adding an organic solvent thereto.

Cyclic-di-AMP sodium salt crystal

Though 3',5&#x2032;-cyclic diadenylic acid was conventionally provided only as a freeze-dried product, a solid material other than a freeze-dried product and a manufacturing method thereof is provided. By a step of adding acid to an aqueous solution of 3&#x2032;,5&#x2032;-cyclic diadenylic acid so as to lower pH to 1 to 3, an inclusion compound of 3&#x2032;,5&#x2032;-cyclic diadenylic acid can be obtained. Said manufacturing method is an extremely simple and easy method and does not need a special machine or the like.

Inclusion compound of 3' ,5&#x2032;-cyclic diadenylic acid and manufacturing method thereof

A crystal of free acid of 3',5&#x2032;-cyclic diguanylic acid containing no metal salt with cobalt, magnesium or the like is provided. A method is sought for obtaining said crystal in a large amount and with ease. By a manufacturing method comprising a step of adding acid to an aqueous solution of 3&#x2032;,5&#x2032;-cyclic diguanylic acid so as to lower pH to 1 to 3, crystals of 3&#x2032;,5&#x2032;-cyclic diguanylic acid can be obtained in a large amount with ease. Said crystals are free acid crystals which do not contain a metal salt with cobalt, magnesium or the like.

Crystalline 3',5&#x2032;-cyclic diguanylic acid

A practical method for enzymatically synthesizing c-di-GMP with excellent productivity is provided. A diguanylate cyclase having physical and chemical characteristics (A) to (F): (A) catalytic action on reaction '2 GTP&#x2192;c-di-GMP'; (B) a molecular weight of 19800&#xb1;2000; (C) an optimum pH of 7.3 to 9.4; (D) an optimum temperature of 35 to 60&#xb0; C.; (E) thermal stability as the remaining activity of 90% or higher after heated for 60 minutes under conditions of 50&#xb0; C. and pH7.8; and (F) the presence of GGDEF (SEQ ID NO:26) domain and the lack of amino acid sequence KXXD (SEQ ID NO:23) in the i-site.

Practical method for enzymatically synthesizing cyclic di-GMP

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

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

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.

Growing community of inventors

Choshi, Japan

Kazuya Ishige