# US 5,707,851 A · Gram-positive alkaliphilic microorganisms

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- Number: US 5,707,851 A
- Document ID: US5707851A
- Status: Granted
- Kind: A
- Application number: 08/314,045
- Filed: 1994-09-28
- Granted: 1998-01-13
- Earliest claimed: 1992-06-24
- Assignees: Genencor International, Inc.
- Inventors: Brian Edward Jones; William Duncan Grant; Nadine Claire Collins
- Web page: https://patentgrep.com/patent/US5707851A

## CPC

- C12N 1/20: Bacteria; Culture media therefor
- C12N 1/205: Bacterial isolates
- C12N 9/00: Enzymes; Proenzymes; Compositions thereof (preparations containing enzymes for cleaning teeth A61K8/66, A61Q11/00; medicinal preparations containing enzymes or proenzymes A61K38/43; enzyme containing detergent compositions C11D; {enzymes with nucleic acid structure, e.g. ribozymes, C12N15/113}); Processes for preparing, activating, inhibiting, separating or purifying enzymes (preparation of malt C12C1/00)
- C12N 9/20: Triglyceride splitting, e.g. by means of lipase
- C12N 9/2417: {from microbiological source}
- C12N 9/52: derived from bacteria {or Archaea}
- C12R 2001/01: Bacteria or Actinomycetales {; using bacteria or Actinomycetales}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

The present invention provides novel aerobic, Gram-positive alkaliphilic bacteria which have been isolated from in and around alkaline soda lakes. These alkaliphiles have been analyzed according to the principles of numerical taxonomy with respect to each other and also to a collection of known bacteria. In addition, these bacterial taxa are further circumscribed by an analysis of the lipid components which serve as chemotaxonomic markers. The alkaliphiles of the present invention produce alkalitolerant enzymes which are capable of performing their functions at high pH which makes them uniquely suited for applications requiring such extreme conditions.

## Claims

### Claim 1

Cells of a single strain of bacteria obtained from a culture consisting of said single strain of bacteria, which are useful for the production of alkalitolerant enzymes, wherein the bacteria consist of aerobic, Gram-positive, short, irregular, rod-shaped, obligate alkaliphilic bacteria having the following characteristics:
a) forms cream-yellow, small, circular colonies;
b) grows optimally above pH 8;
c) gives a positive response to the following tests:
1) Gelatin hydrolysis
2) Galactose
3) .beta.-glucosidase
4) Arginine
5) Bacitracin
6) N-acetyl-D-glucosamine
7) Cellobiose
8) Inosine
9) Uridine; and
d) gives a negative response to the following tests:
1) Maltose
2) Acetate
3) D-glucose
4) D-melibiose
5) Propionate
6) Valerate
7) Glycogen
8) Serine
9) Ampicillin
10) Penicillin G
11) Methicillin
12) Tetracyclin
13) Methyl pyruvate
14) Mono-methylsuccinate
15) .alpha.-ketobutyric acid.

### Claim 2

Cells of a single strain of bacteria obtained from a culture consisting of said single strain of bacteria, which are useful for the production of alkalitolerant enzymes, wherein the bacteria consist of aerobic, Gram-positive, coryneform, obligate alkaliphilic bacteria having the following characteristics:
a) mature colonies that are glistening, bright orange/red;
b) grows optimally between about pH 9 and 10;
c) gives a positive response to the following tests:
1) Arginine
2) Methionine
3) Glycine
4) Valine
5) Methicillin
6) Tetracyclin
7) Bacitracin
8) .alpha.-cyclodextrin
9) Tween 40
10) Methyl pyruvate
11) Mono-methylsuccinate
12) .beta.-hydroxybutyric acid
13) Bromo-succinic acid; and
d) gives a negative response to the following tests:
1) Gelatin hydrolysis
2) Fumarate
3) Fructose
4) Galactose
5) D-saccharose
6) D-melibiose
7) Glycogen
8) L-serine
9) .beta.-glucosidase
10) N-acetyl-D-glucosamine
11) Cellobiose
12) D-sorbitol
13) D-trehalose
14) Turanose.

### Claim 3 (depends on 1)

A pure culture consisting of the single strain of bacteria of claim 1.

### Claim 4 (depends on 2)

A pure culture consisting of the single strain of bacteria of claim 2.