Gene encoding lacto-N-biosidase

• US 5,908,772 A
• Filed: 04/26/1996
• Issued: 06/01/1999
• Est. Priority Date: 04/27/1995
• 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: Mitta, Masanori, Kato, Ikunoshin, Sano, Mutsumi
• Primary Examiner(s): Prouty, Rebecca E.
• Application Number:
  US08/637,899
• Time in Patent Office: 1,131 Days
• Field of Search: 536/23.2, 536/23.7, 536/23.1, 435/71.1, 435/71.2, 435/200, 435/172.3, 435/252.3, 435/240.2, 435/320.1, 435/325, 435/419
• US Class Current: 435/200
• CPC Class Codes: C12N 9/2402 hydrolysing O- and S- glycosyl compounds (3.2.1)
• CPC Class Codes: C12Y 302/0114 Lacto-N-biosidase (3.2.1.140)

Abstract

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

Claims

1. An isolated DNA encoding a polypeptide possessing lacto-N-biosidase activity, the isolated DNA being selected from the group consisting of:

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

1;

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

2; and

(c) a DNA which hybridizes to (b) above at 65°

C. for 20 hours in a solution containing 6×

SSC, 1% sodium lauryl sulfate, 100 μ

g/ml salmon sperm DNA and 5×

Denhardt'"'"'s solution.

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

3. The isolated DNA according to claim 2, wherein the strain is Streptomyces sp 142 (FERM BP-4569).

4. The isolated DNA according to claim 1, wherein said DNA encodes a polypeptide of SEQ ID NO:

1.

5. The isolated DNA according to claim 4, having a DNA sequence of SEQ ID NO:

2.

6. An expression vector which comprises the DNA as defined in claim 1, wherein the expression vector propagates in a procaryotic or eucaryotic cell.

7. The expression vector according to claim 6, wherein the polypeptide encoded by the DNA has the amino acid sequence of SEQ ID NO:

1.

8. The expression vector according to claim 6, containing a DNA sequence of SEQ ID NO:

2.

9. A cell of a procaryote or eucaryote transformed with an expression vector as defined in claim 6.

10. The cell according to claim 9, wherein a polypeptide encoded by the expression vector has the amino acid sequence of SEQ ID NO:

1.

11. The cell according to claim 9, wherein the expression vector contains a sequence of SEQ ID NO:

2.

12. The cell according to claim 9, wherein said eucaryote or said procaryote is selected from the group consisting of microorganisms, animal cells and plant cells.

13. The cell according to claim 9, wherein said eucaryote is Escherichia coli.

14. A method for producing a polypeptide possessing lacto-N-biosidase activity comprising the steps of:

(a) culturing the cell of claim 9; and

(b) recovering the polypeptide possessing lacto-N-biosidase activity from the culture obtained in Step (a).

15. A synthetic oligonucleotide probe or primer which specifically hybridizes with the isolated DNA of claim 1 at 65°

C. for 20 hours in a solution containing 6×

SSC, 1% sodium lauryl sulfate, 100 μ

g/ml salmon sperm DNA and 5×

Denhardt'"'"'s solution.