# US 2023/0057586 A1 · Optimized culture media for clostridia bacteria

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- Number: US 2023/0057586 A1
- Document ID: US20230057586A1
- Status: Published application
- Kind: A1
- Application number: 17/633,996
- Filed: 2020-07-21
- Published: 2023-02-23
- Earliest claimed: 2019-08-16
- Assignees: Janssen Biotech, Inc.
- Applicants: Janssen Biotech, Inc.
- Inventors: John Bodek; Kristina Demjanick; Laura Grussendorf; John Jerbasi; Claus Lang; Dominik Lüthy; Peter Hebbeln; Shant Shahinian; Stuti Mehta; Susan Lenk
- Web page: https://patentgrep.com/patent/US20230057586A1

## CPC

- C12N 1/00: Microorganisms; Compositions thereof (medicinal preparations containing material from protozoa, bacteria or viruses A61K35/66, from algae A61K36/02, from fungi A61K36/06; preparing medicinal bacterial antigen or antibody compositions, e.g. bacterial vaccines, A61K39/00); Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N 1/205: Bacterial isolates
- C12R 2001/145: Clostridium
- C12R 2001/45: Staphylococcus epidermidis
- 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

Toxigenic Clostridium spoilage culture comprises 1.5-2 pts. wt. peptone, 1.5-2 pts. wt. casein, 0.5-0.75 pts. wt, yeast extract powder, 0.00014-0.00028 pts. wt. zinc sulfate heptahydrate, 0.5-0.75 pts. wt, sodium dihydrogen phosphate dodecahydrate, 0.03-0.045 pts, wt. potassium dihydrogen phosphate and 1-1.5 pts. wt. glucose.

## Claims

### Claim 1

A medium comprising:
(a) between 45-55 g/kg of HY-Peptone,
(b) between 15-25 g/kg of Yeast extract,
(c) between 9-14 g/kg of M9 salts,
(d) between 5-15 g/kg of glucose,
wherein the medium is for culturing a bacterial strain belonging to Clostridium genus.

### Claim 2 (depends on 1)

The medium of claim 1, wherein the medium comprises:
(a) 50 g/kg of HY-Peptone,
(b) 20 g/kg of Yeast extract,
(c) 11.4 g/kg of M9 salts,
(d) 10 g/kg of glucose.

### Claim 3 (depends on 1, 2)

The medium of claim 1 or 2, wherein the bacterial strain belonging to Clostridium genus is a bacterial strain belonging to Clostridium clusters IV or XIVa.

### Claim 4 (depends on 1, 2)

The medium of claim 1 or 2, wherein the bacterial strain belonging to Clostridium genus is selected from the group consisting of the strains listed in Table 1.

### Claim 5 (depends on 1, 2)

The medium of claim 1 or 2, wherein the bacterial strain belonging to Clostridium genus comprises a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 17.

### Claim 6 (depends on 1)

The medium of claim 1, wherein the medium is maintained in anaerobic conditions.

### Claim 7 (depends on 1)

The medium of claim 1, wherein the medium further comprises at least one bacterium comprising 16S rDNA sequences selected from the group: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 17.

### Claim 8 (depends on 1, 2)

A process for culturing bacterial cells, said process comprising:
a) providing a bioreactor,
b) mixing the cells to be cultured with the medium according to claim 1 or 2,
c) incubating the resultant mixture.

### Claim 9 (depends on 8)

The process of claim 8, wherein the bacterial strain belonging to Clostridium genus is a bacterial strain belonging to Clostridium clusters IV or XIVa.

### Claim 10 (depends on 8)

The process of claim 8, wherein the bacterial strain belonging to Clostridium genus is selected from the group consisting of the strains listed in Table 1.

### Claim 11 (depends on 8)

The process of claim 8, wherein the bacterial strain belonging to Clostridium genus comprises a sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, SEQ ID NO: 8, SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, and SEQ ID NO: 17.

### Claim 12 (depends on 8)

The process of claim 8, wherein the medium has an initial pH of between about 5.8-7.

### Claim 13 (depends on 8)

The process of claim 8, wherein the medium is maintained in anaerobic conditions.