Glucoamylase gene of rhizopus oryzae

• US 4,863,864 A
• Filed: 12/13/1985
• Issued: 09/05/1989
• Est. Priority Date: 12/15/1984
• Status: Expired due to Term

• Current Assignee: Suntory Ltd (Kotobuki Realty Co., Ltd.)
• Original Assignee: Suntory Ltd (Kotobuki Realty Co., Ltd.)
• Inventors: Tanaka, Yoshikazu, Nakamura, Norihisa, Ashikari, Toshihiko, Shibano, Yuji, Yoshizumi, Hajime
• Primary Examiner(s): Weimar, Elizabeth C.
• Assistant Examiner(s): Ellis, Joan
• Application Number:
  US06/808,742
• Time in Patent Office: 1,362 Days
• Field of Search: 435/172.3, 435/253, 435/91, 435/254, 435/255, 435/320, 435/68, 435/942, 435/205, 536/27, 935/18, 935/19, 935/28, 935/37, 935/41, 935/60, 935/69
• US Class Current: 435/205
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C12N 9/2428 Glucan 1,4-alpha-glucosidase (3.2.1.3), i.e. glucoamylase
• CPC Class Codes: Y10S 435/942 Saccharomyces cerevisiae

Abstract

The present invention relates to the Rhizopus derived glucoamylase gene, a novel recombinant vector comprising said gene, and a microorganism transformed by said vector, as well as a process for reproducing Rhizopus glucoamylase by cultivating the transformed microorganism, especially yeast, in a liquid medium.

Claims

1. The Rhizopus oryzae glucoamylase structural gene consisting of the following nucleotide sequence:

##STR1##

3. The recombinant DNA of claim 1, operatively-linked to the Rhizopus oryzae glucoamylase gene signal sequence consisting of the following nucleotide sequence:

space="preserve" listing-type="tabular">__________________________________________________________________________ATG CAA CTG TTC AAT TTG CCA TTG AAA GTT TCA TTC TTT CTC GTC CTC TCT TAC TTT TCT TTG CTC GTT TCT GCT. __________________________________________________________________________

5. A recombinant vector comprising the glucoamylase structural gene of claim 1.

6. A recombinant vector comprising the glucoamylase gene of claim 3.

8. A microorganism transformed by the vector of claim 5.

9. A microorganism transformed by the vector of claim 6.

11. A process for producing glucoamylase of the genus Rhizopus oryzae capable of efficient hydrolysis of raw starch, said process comprising the steps of:

(a) transforming Saccharomyces cerevisae yeast cells with a recombinant expression vector which comprises the glucoamylase structural gene of claim 1;

(b) cultivating said yeast cells in a liquid medium capable of inducing the expression of the glucoamylase structural gene; and

,

(c) recovering said glucoamylase from the medium or cells.

12. A process according to claim 11, wherein said glucoamylase structural gene is operatively linked to the Rhizopus oryzae glucoamylase signal peptide-encoding sequence, wherein cultivation is carried out under conditions which enable the glucoamylase to be secreted from said yeast cells into the culture medium from which medium the glucoamylase is then recovered.

2. The Rhizopus oryzae glucoamylase structural gene wherein the gene encodes the following amino acid sequence:

##STR2##

4. The recombinant DNA of claim 2 which encodes the following amino acid sequence:

##STR3##

7. A plasmid selected from the group consisting of pCGA449, pCGA469, pYGA2249, pYGA2149, pYGA2269 and pYGA195.

10. A microorganism transformed by plasmid pCGA449, pCGA469, pYGA2169, pYGA2149, pYGA2269 or pYGA195.