Gene encoding endo-.beta.-N-acetylglucosaminidase A

• US 5,985,637 A
• Filed: 11/19/1997
• Issued: 11/16/1999
• Est. Priority Date: 10/27/1995
• Status: Expired due to Fees

• Current Assignee: Takara Shuzo Company Limited (Ningbo Junsheng Investment Group Co., Ltd.)
• Original Assignee: Takara Shuzo Company Limited (Ningbo Junsheng Investment Group Co., Ltd.)
• Inventors: Takegawa, Kaoru, Iwahara, Shojiro
• Primary Examiner(s): Wax, Robert A.
• Assistant Examiner(s): Saidha, Tekchand
• Application Number:
  US08/969,714
• Time in Patent Office: 727 Days
• Field of Search: 435/200, 435/201, 435/205, 435/69.1, 435/252.3, 435/320.1, 435/830, 530/350, 536/23.2, 536/23.7
• US Class Current: 435/200
• CPC Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12Y 302/01096 Mannosyl-glycoprotein endo-beta-N-acetylglucosaminidase (3.2.1.96)

Abstract

An isolated DNA having a sequence encoding a polypeptide possessing endo-β-N-acetylglucosaminidase A activity or functionally equivalent variants thereof; and a method for producing a polypeptide possessing endo-β-N-acetylglucosaminidase A activity or functionally equivalent variants thereof using the isolated DNA by recombinant DNA technology.

Claims

1. An isolated DNA encoding a polypeptide possessing endo-β

-N-acetylglucosaminidase A activity, the isolated DNA being selected from the group consisting of;

(a) a DNA having the nucleotide sequence of SEQ ID NO;

2;

(b) a DNA having a nucleotide sequence encoding the amino acid sequence of SEQ ID NO;

1; and

(c) a DNA sequence hybridizing to (a) or (b) under conditions of using a solution containing 6×

SSC, 0.5% SDS, 5×

Denhardt'"'"'s solution and 100 μ

g/ml of salmon sperm DNA, and incubating overnight at 65°

C.

2. The isolated DNA according to claim 1, wherein the polypeptide is derived from a strain of the genus Arthrobacter.

3. The isolated DNA according to claim 2, wherein the polypeptide is derived from Arthrobacter protoformiae AKU 0647 strain (FERM BP-4948).

4. A recombinant DNA which comprises a DNA sequence of any one of claims 1 to 3.

5. A vector which comprises the recombinant DNA of claim 4.

6. The vector according to claim 5, wherein said recombinant DNA is operably linked to a promoter.

7. A cell of a prokaryote or eucaryote transformed with the vector of claim 5.

8. A method for producing a polypeptide possessing endo-β

-N-acetylglucosaminidase A activity, comprising the steps of;

(a) culturing the cell of claim 7; and

(b) recovering the polypeptide possessing endo-β

-N-acetylglucosaminidase A activity from the culture obtained in Step (a).