Meiji Seika Kaisha, Ltd.

Koichi Miyamoto

Takeshi Murakami

Naoki Midoh

Manabu Watanabe

Koji Yanai

Kaoru Okakura

Horst Kleinkauf

Naomi Sumida

Rainer Zocher

Shohei Yasuda

Sato Aihara

Tetsuya Yasutake

Takafumi Futamura

The present invention provides an enzyme that synthesizes a cyclic depsipeptide, particularly the substance 1022, and a gene thereof. A cyclic depsipeptide synthetase according to the present invention comprises (a) an amino acid sequence of SEQ ID NO: 2 or (b) a modified amino acid sequence of the amino acid sequence of SEQ ID NO: 2 that have one or more modifications selected from a substitution, a deletion, an addition and an insertion and has cyclic depsipeptide synthetase activity. A cyclic depsipeptide synthetase gene according to the present invention comprises a nucleotide sequence encoding a cyclic depsipeptide synthetase. The present invention also provides a recombinant vector and a transformant for expressing the cyclic depsipeptide synthetase, and a mass production system for the cyclic depsipeptide. The present invention further provides a method for producing the cyclic depsipeptide synthetase.

Cyclic depsipeptide synthetase and method for recombinant production

An objective of the present invention is to provide a transformant altered so as to produce a secondary metabolite in which a benzene ring of the secondary product is modified at the para-position with a functional group containing a nitrogen atom. A transformant according to the present invention is a transformant of an organism producing a secondary metabolite having a benzene ring skeleton without substitution at the para-position with a functional group containing a nitrogen atom, said transformant being transformed by introducing genes involved in a biosynthetic pathway from chorismic acid to p-aminophenylpyruvic acid, including a gene encoding an amino acid sequence (SEQ ID NO: 2) having 4-amino-4-deoxychorismic acid synthase activity, a gene encoding an amino acid sequence (SEQ ID NO: 4) having 4-amino-4-deoxychorismic acid mutase activity and a gene encoding an amino acid sequence (SEQ ID NO: 6) having 4-amino-4-deoxyprephenic acid dehydrogenase activity, so as to produce a secondary metabolite having a benzene ring skeleton substituted at the para-position with a functional group containing a nitrogen atom. Another objective of the present invention is to provide a novel gene involved in the biosynthetic pathway from chorismic acid to p-aminophenylpyruvic acid. A novel gene according to the present invention comprises genes encoding the amino acid sequences of SEQ ID NOs: 2, 4 and 6 or modified sequences thereof.

Transformant producing secondary metabolite modified with functional group and novel biosynthesis genes

An objective of the present invention is to provide a D-phenyllactic acid dehydrogenase. Another objective of the present invention is to provide a gene which encodes the D-phenyllactic acid dehydrogenase. A D-phenyllactic acid dehydrogenase according to the present invention is a protein comprising an amino acid sequence of the amino acid sequence of SEQ ID NO: 2 or a modified amino acid sequence of the amino acid sequence of SEQ ID NO: 2 that has one or more modifications selected from a substitution, a deletion, an addition and an insertion and has D-phenyllactic acid dehydrogenase activity. A D-phenyllactic acid dehydrogenase gene according to the present invention comprises a nucleotide sequence which encodes the D-phenyllactic acid dehydrogenase, for example the nucleotide sequence of SEQ ID NO: 1.

(R)-2-hydroxy-3-phenylpropionate (d-phenyllactate) dehydrogenase and gene encoding the same

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Odawara, Japan

Koichi Miyamoto