# US 2014/0193854 A1 · Method for Mass-Producing Antifreeze Protein Derived from Polar Yeast

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- Number: US 2014/0193854 A1
- Document ID: US20140193854A1
- Status: Published application
- Kind: A1
- Application number: 14/117,225
- Filed: 2011-05-17
- Published: 2014-07-10
- Earliest claimed: 2011-05-13
- Assignees: Korean Ocean Research & Development Institute
- Inventors: Hakjun Kim; Sungho Kang; Junhyuck Lee; Sunggu Lee; Sejong Han; Jongchan Park; Kyoungsun Park
- Web page: https://patentgrep.com/patent/US20140193854A1

## CPC

- C07K 14/39: from yeasts
- C12N 15/815: {for yeasts other than Saccharomyces}
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present invention relates to a method for mass-producing an antifreeze protein derived from a polar yeast, and more particularly, to a method for mass-producing an antifreeze protein derived from Leucosporidium sp., which is the polar yeast, for synthesizing a recombinant polynucleotide by optimizing and altering a gene, which codes the antifreeze protein derived from the polar yeast, for a yeast expression system, and for expressing same using the yeast expression system.

## Claims

### Claim 1

A method for mass-producing an antifreeze protein derived from arctic yeast or a culture medium containing the same, the method comprising the steps of:
(1) synthesizing a polynucleotide comprising the nucleotide sequence of SEQ. ID. NO: 1;
(2) introducing the polynucleotide of step (1) into a Pichia pastoris-derived expression vector;
(3) culturing a yeast transformant transformed with the recombinant expression vector; and
(4) obtaining a culture medium from step (3).

### Claim 2 (depends on 1)

The method of claim 1, wherein the Pichia pastoris-derived expression vector in step (2) is pPICZαA.

### Claim 3 (depends on 1)

The method of claim 1, wherein, the recombinant expression vector in step (3) is pPICZαA-AY30 (Accession No.: KCTC 11917BP).

### Claim 4

A polynucleotide comprising the nucleotide sequence of SEQ. ID. NO: 1, expressing an antifreeze protein.