# US 5,705,616 A · Proteins from yeast host strains with defects in N-glycosylation

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 5,705,616 A
- Document ID: US5705616A
- Status: Granted
- Kind: A
- Application number: 08/461,593
- Filed: 1995-06-05
- Granted: 1998-01-06
- Earliest claimed: 1992-08-07
- Assignees: Boehringer Mannheim GmbH
- Inventors: Ludwig Lehle; Klaus Lehnert; Erhard Kopetzki
- Web page: https://patentgrep.com/patent/US5705616A

## CPC

- C12N 9/0006: {acting on CH-OH groups as donors (1.1)}
- C12N 1/16: Yeasts; Culture media therefor
- C12N 1/185: Saccharomyces isolates
- C12P 21/005: {Glycopeptides, glycoproteins}
- C12R 2001/85: Saccharomyces
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12P: FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE {(brewing of beer C12C; producing vinegar C12J; producing specific peptides or proteins C07K; producing enzymes C12N9/00; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification C12N15/00; measuring or testing processes involving enzymes or microorganisms C12Q; measuring or testing processes involving nucleic acid amplification reactions C12Q1/6844; fermentation processes to form a food composition, A21 or A23; compounds in general, see the relevant compound class, e.g. C01, C07)}
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

Saccharomyces mutants with defects in N-glycosylation which are obtainable by ›.sup.3 H!-mannose suicide selection, introduction of one or several selective markers, selection of those strains which, after transformation with the plasmid YEpL/glucose oxidase, secrete 10 mg/l GOD or more into the culture medium after culture under standard conditions, are allelic to the ngd mutations in Saccharomyces cerevisiae, DSM 7042, DSM 7338, DSM 7160 and/or 7340 and express proteins with a uniform carbohydrate structure.

## Claims

### Claim 1

A hypoglycosylated recombinant protein obtained by transformation of a Saccharomyces cerevisiae mutant with DNA which codes for the desired protein, said mutant being of a phenotype which is not temperature sensitive, said mutant having defects in N-glycosylation resulting from alteration in the ngd 29 and/or ngd 62 genes and yielding a diploid having defects in the N-glycosylation when crossed with a strain of Saccharomyces cerevisiae selected from the group consisting of DSM 7042, DSM 7338, DSM 7160, and DSM 7340, fermentation of the cells of said transformed mutant, and isolation of said protein from the cells or from the culture supernatant, wherein said protein would be hyperglycosylated if expressed in wild-type Saccharomyces cerevisiae.

### Claim 2 (depends on 1)

The protein of claim 1 wherein the mutant yields a diploid having defects in N-glycosylation when crossed with DSM 7042.

### Claim 3 (depends on 1)

The protein of claim 1 wherein the mutant yields a diploid having defects in N-glycosylation when crossed with DSM 7338.

### Claim 4 (depends on 1)

The protein of claim 1 wherein the mutant yields a diploid having defects in N-glycosylation when crossed with DSM 7160.

### Claim 5 (depends on 1)

The protein of claim 1 wherein the mutant yields a diploid having defects in N-glycosylation when crossed with DSM 7340.