# US 5,346,891 A · Lipopolysaccharide-producing bacteria, lipopolysaccharides, and lipopolysaccharide-containing, medicines and veterinary medicines

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- Number: US 5,346,891 A
- Document ID: US5346891A
- Status: Granted
- Kind: A
- Application number: 07/747,633
- Filed: 1991-08-20
- Granted: 1994-09-13
- Earliest claimed: 1990-08-20
- Assignees: Gen-Ichiro Soma; Den'ichi Mizuno
- Inventors: Gen-Ichiro Soma; Kiyoshi Yoshimura; Daisuke Tsukioka; Den'ichi Mizuno; Haruyuki Oshima
- Web page: https://patentgrep.com/patent/US5346891A

## CPC

- C12N 1/205: Bacterial isolates
- A61K 35/74: Bacteria (therapeutic use of a bacterial protein A61K38/00)
- A61K 45/05: {Immunological preparations stimulating the reticulo-endothelial system, e.g. against cancer}
- C12P 19/04: Polysaccharides, i.e. compounds containing more than five saccharide radicals attached to each other by glycosidic bonds
- C12R 2001/01: Bacteria or Actinomycetales {; using bacteria or Actinomycetales}
- C12R 2001/425: Serratia
- Y02A 50/30: Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- 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
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Novel LPS having the following physical properties, novel bacilli providing those LPS, and novel immunity stimulators, anagesics, and antiwithdrawal agents, and novel veterinary immunity stimulators, analgesics, and antiwithdrawal agents containing LPS selected from the group consisting of those LPSs and mixtures thereof. LPS1 Dominant molecular weight: 5,000.+-.1,000 as determined by ______________________________________ SDS-PAGE method ______________________________________ Phosphorus number: 2 .+-. 1/5,000 (m.w.) Hexosamine number: 9 .+-. 1/5,000 (m.w.) KDO number: 2 .+-. 1/5,000 (m.w.) ______________________________________ LPS2 Dominant molecular weight: 6,500.+-.2,500 as determined by ______________________________________ SDS-PAGE method ______________________________________ Phosphorus number: 1 to 2/5,000 (m.w.) Hexosamine number: 7 .+-. 1/5,000 (m.w.) KDO number: 1 to 2/5,000 (m.w.) ______________________________________ LPS3 Dominant molecular weight: 6,500.+-.2,500 as determined by ______________________________________ SDS-PAGE method ______________________________________ Phosphorus number: 2 .+-. 1/5,000 (m.w.) Hexosamine number: 5 .+-. 1/5,000 (m.w.) KDO number: 2 .+-. 1/5,000 (m.w.) ______________________________________

## Claims

### Claim 1

A lipopolysaccharide produced by a strain of the species Serratia ficaria, having all of the identifying characteristics of FERM BP-3509 and exhibiting a dominant molecular weight of 5,000.+-.1,000 as determined by SDS-PAGE method, having 2.+-.1 phosphorus, 9.+-.1 hexoeamines and 2.+-.1 KDO per molecular weight of 5,000, wherein the identifying characteristics are as follows:
a) Morphological characteristics
1) Small rod
2) No Motility
3) Gram stain: -
b) Growth
1) Standard agar medium: A yellow to creamy round opaque colony is formed;
2) SS agar medium: A white translucent colony is formed;
3) TSI agar medium: No change is found on the slant, but a higher layer changes to yellow; Gas is produced.
c) Physiological characteristics
1) Voges-Proskauer reaction: +
2) Indole production: -
3) Hydrogen sulfide production: -
4) Utilization of citrate: +
5) Urease: -
6) Oxidass: -
7) O-F test: +
d) Utilization of carbon sources
1) Lactose: +
2) Adonitol: -
3) Rhamnose: +
4) Mannitol: +
5) Esculin: +
6) Inositol: -
7) Sorbitol: +
8) Arabinose: +
9) Raffinose: +
10) Sucrose: +
e) Others
1) Lyein decarboxylase: -
2) Utilization of malonate: -
3) Arginine dihydroxylase: -
4) Phenylalanine deaminase: -
5) Ornithine decarboxylase: -

### Claim 2

A lipopolysaccharide produced by a strain of the species Enterobacter cloacae, having all of the identifying characteristics of FERM BP-3510 and exhibiting a dominant molecular weight of 6,500.+-.2,500 as determined by SDS-PAGE method, having 1-2 phosphorus, 7.+-.1 hexosamines and 1-2 KDO per molecular weight of 5,000, wherein the identifying characteristics are as follows:
a) Morphological characteristics
1) Small rod
2) No Motility
3) Gram stain: -
b) Growth
1) Standard agar medium: A creamy opaque colony is formed;
2) SS agar medium: A red opaque colony is formed;
3) TSI agar medium: No change is found on the slant, but a higher layer changes to yellow; Gas is produced;
c) Physiological characteristics
1) voges-Proskauer reaction: +
2) Indole production: -
3) Hydrogen sulfide production: -
4) Utilization of citrate: +
5) Urease: -
6) Oxidase: -
7) O-F test: +
d) Utilization of carbon sources
1) Lactose: +
2) Adonitol: -
3) Rhamnose: +
4) Mannitol: +
5) Esculin: +
6) Inositol: -
7) Sorbitol: +
8) Arabinose: +
9) Raffinose: +
10) Sucrose: +
e) Others
1) Lyein decarboxylase: -
2) Utilization of malonate: +
3) Arginine dihydroxylase: +
4) Phenylalanine deaminase: -
5) Ornithine decarboxylase: +

### Claim 3

A lipopolysaccharide produced by a strain of the species Pantoea agglomerans, having all of the identifying characteristics of FERM BP-3511 and exhibiting a dominant molecular weight of 6,500.+-.2,500 as determined by SDS-PAGE method, having 2.+-.1 phosphorus, 5.+-.1 hexosamines and 2.+-.1 KDO per molecular weight of 5,000, wherein the identifying characteristics are as follows:
a) Morphological characteristics
1) Small rod
2) No Motility
3) Gram stain: -
b) Growth
1) Standard agar medium: A yellow round translucent colony is formed;
2) SS agar medium: No colony is formed;
3) TSI agar medium: No change is found on the slant, but a higher layer changes to yellow. Gas is not produced;
c) Physiological characteristics
1) Voges-Proskauer reaction: +
2) Indole production: -
3) Hydrogen sulfide production: -
4) Utilization of titrate: +
5) Urease: -
6) Oxidass: -
7) O-F test: +
d) Utilization of carbon sources
1) Lactose: +
2) Adonitol: -
3) Rhamnose: +
4) Mannitol: +
5) Esculin: +
6) Inositol: -
7) Sorbitol: +
8) Arabinose: +
9) Raffinose: -
10) Sucrose: +
e) Others
1) Lysin decarboxylase: -
2) Utilization of malonate: +
3) Arginine dihydroxylase: -
4) Phenylalanine deaminase: -
5) Ornithine decarboxylase: -

### Claim 4 (depends on 1)

An immunostimulant comprising the lipopolysaccharide of claim 1, together with a pharmaceutically or veterinarily acceptable carrier.

### Claim 5 (depends on 2)

An immunostimulant comprising the lipopolysaccharide of claim 2, together with a pharmaceutically or veterinarily acceptable carrier.

### Claim 6 (depends on 3)

An immunostimulant comprising the lipopolysaccharide of claim 3, together with a pharmaceutically or veterinarily acceptable carrier.