Amidase gene

Details Amidase gene Amidase gene

2001-10-16

2003-09-09

Slobodyansky, Elizabeth (Department: 1652)

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing alpha or beta amino acid or substituted amino acid...

C435S116000, C435S136000, C435S139000, C435S228000, C435S252300, C435S252330, C435S280000, C435S320100, C536S023200

Reexamination Certificate

active

06617139

ABSTRACT:

CROSS REFERENCE TO RELATED APPLICATION
The present application is a 371 application of PCT/JP00/02492 filed Apr. 17, 2000.
FIELD OF THE INVENTION
The present invention relates to a novel protein having amidase activity that stereoselectively hydrolyzes &agr;-amino acid amides and &agr;-hydroxy acid amides, and a gene encoding said protein.
BACKGROUND OF THE PRESENT INVENTION
It is known that optically active &agr;-amino acids and &agr;-hydroxy acids can be produced using microorganism or enzyme threof as a catalyst to hydrolyze &agr;-amino acid amides and &agr;-hydroxy acid amides. Examples of optically active &agr;-amino acids are described in Japanese Patent Application Laying-Open (kokai) Nos. 61-293394 and 62-55097; optically active &agr;-hydroxy acids are described in Japanese Patent Application Laying-Open (kokai) No. 2-84198.
SUMMARY OF THE INVENTION
Amidase genes cloned via gene recombination techniques can greatly enhance the catalytic capability of microorganisms compared with the conventional methods, because it enables the existence of multiple copies of amidase genes in a cell.
In order to prepare a microbial catalyst having higher catalytic activity, we have completed the present invention by cloning an amidase gene which stereoselectively hydrolyzes &agr;-amino acid amides and &agr;-hydroxy acid amides.
The present invention provides the followings:
(1) A protein defined in the following (a) or (b):
(a) a protein having an amino acid sequence of SEQ ID NO: 2;
(b) a protein having an amino acid sequence that differs from the amino acid sequence of SEQ ID NO: 2 by deletion, replacement or addition of one or more amino acids, and having amidase activity that stereoselectively hydrolyzes &agr;-amino acid amides and &agr;-hydroxy acid amides.
(2) A gene which encodes the protein of (1)
(3) The gene of (2) which having a nucleotide sequence of nucleotides 361 to 1305 of SEQ ID NO: 1.
(4) A gene capable of hybridizing to the nucleotide sequence of nucleotides 365 to 1305 of SEQ ID NO: 1 or fragments thereof, and encodes a protein having amidase activity that stereoselectively hydrolyzes &agr;-amino acid amides and &agr;-hydroxy acid amides.
(5) A recombinant vector to which the gene of any one of (2) to (4) is linked.
(6) A transformant which is prepared by introducing the recombinant vector of (5) into a host microorganism.
(7) A method for producing a protein having amidase activity which comprises a culturing process of the transformant of (6)
When a protein having amidase activity, which has been obtained by culturing the above transformant, acts on &agr;-amino acid amides, optically active &agr;-amino acids corresponding thereto can be produced; and when the same protein acts on &agr;-hydroxy acid amides, optically active &agr;-hydroxy acids corresponding thereto can be produced.
This specification includes part or all of the contents disclosed in the specification and/or drawings of Japanese Patent Application No. 11-109328, which is a priority document of the present application.


REFERENCES:
patent: 5215897 (1993-06-01), Sakashita et al.
patent: 5248608 (1993-09-01), Van Dooren et al.
patent: 5766918 (1998-06-01), Petre et al.
patent: 5888785 (1999-03-01), Fallon et al.
patent: WO 97/12964 (1997-04-01), None
patent: WO 98/01568 (1998-01-01), None
E. Mahenthiralingam, et al., Journal of General Microbiology, vol. 139, No. 3, pp. 575-583, “Cloning and Sequencing of the Gene Which Encodes the Highly Inducible Acetamidase of Mycobacterium Smegmatis”, 1993.

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