# US 10,767,141 B2 · Thermolysin for easy-cleaning of insect body stains

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- Number: US 10,767,141 B2
- Document ID: US10767141B2
- Status: Granted
- Kind: B2
- Application number: 15/193,242
- Filed: 2016-06-27
- Published: 2016-10-20
- Granted: 2020-09-08
- Earliest claimed: 2010-06-21
- Assignees: Toyota Motor Engineering & Manufacturing North America, Inc.; Toyota Motor Corporation
- Applicants: Toyota Motor Engineering & Manufacturing North America, Inc.; Toyota Motor Corporation
- Inventors: Songtao Wu; Hongfei Jia; Masahiko Ishii; Minjuan Zhang
- Web page: https://patentgrep.com/patent/US10767141B2

## CPC

- C11D 3/386: Preparations containing enzymes {, e.g. protease or amylase}
- C08L 33/00: Compositions of homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and only one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides or nitriles thereof; Compositions of derivatives of such polymers
- C08L 89/00: Compositions of proteins; Compositions of derivatives thereof
- C09D 5/1637: {Macromolecular compounds}
- C09D 5/1662: {Synthetic film-forming substance}
- C09D 133/00: Coating compositions based on homopolymers or copolymers of compounds having one or more unsaturated aliphatic radicals, each having only one carbon-to-carbon double bond, and at least one being terminated by only one carboxyl radical, or of salts, anhydrides, esters, amides, imides, or nitriles thereof; Coating compositions based on derivatives of such polymers
- C09D 189/00: Coating compositions based on proteins; Coating compositions based on derivatives thereof
- C11D 3/04: Water-soluble compounds
- C11D 3/044: {Hydroxides or bases}
- C11D 3/2003: {Alcohols; Phenols}
- C11D 3/37: Polymers {(polymeric surface-active agents C11D1/008; polysaccharides C11D3/222)}
- C11D 3/3757: {(Co)polymerised carboxylic acids, -anhydrides, -esters in solid and liquid compositions}
- C11D 3/3769: {(Co)polymerised monomers containing nitrogen, e.g. carbonamides, nitriles or amines}
- C11D 3/38618: {Protease or amylase in liquid compositions only}
- C11D 7/04: Water-soluble compounds
- C11D 7/261: {Alcohols; Phenols}
- C12Y 304/24027: Thermolysin (3.4.24.27)
- C11D 2111/18: Glass; Plastics
- C11D: DETERGENT COMPOSITIONS; USE OF SINGLE SUBSTANCES AS DETERGENTS; SOAP OR SOAP-MAKING; RESIN SOAPS; RECOVERY OF GLYCEROL
- C08L: COMPOSITIONS OF MACROMOLECULAR COMPOUNDS (compositions based on polymerisable monomers C08F, C08G; artificial filaments or fibres D01F; textile treating compositions D06)
- C09D: COATING COMPOSITIONS, e.g. PAINTS, VARNISHES OR LACQUERS; FILLING PASTES; CHEMICAL PAINT OR INK REMOVERS; INKS; CORRECTING FLUIDS; WOODSTAINS; PASTES OR SOLIDS FOR COLOURING OR PRINTING; USE OF MATERIALS THEREFOR (cosmetics A61K; processes for applying liquids or other fluent materials to surfaces, in general, B05D; staining wood B27K5/02; glazes or vitreous enamels C03C; natural resins, French polish, drying-oils, driers, turpentine, per se, C09F; polishing compositions other than French polish, ski waxes C09G; adhesives or use of materials as adhesives C09J; materials for sealing or packing joints or covers C09K3/10; materials for stopping leaks C09K3/12; processes for the electrolytic or electrophoretic production of coatings C25D)
- C12Y: ENZYMES

## Abstract

A substrate or coating is provided that includes a protease with enzymatic activity toward a component of a biological stain. Also provided is a process for facilitating the removal of a biological stain is provided wherein an inventive substrate or coating including a protease is capable of enzymatically degrading of one or more components of the biological stain to facilitate biological stain removal from the substrate or said coating.

## Claims

### Claim 1

A method of facilitating the removal of a biological stain on a substrate or a coating comprising:
providing a liquid coating material comprising an alcohol selected from the group consisting methanol, ethanol, isopropyl alcohol, propanol and butanol, wherein the concentration of said alcohol in said liquid coating is from 20 to 40% wt. %, wherein the pH of the liquid coating is geater than 5.0, and associating a themolysin-like protease, wherein the protease has an activity in excess of 20,000 U/mg, with said liquid coating material to form a liquid bioactive coating material such that said protease is capable of enzymatically degrading a component of a biological stain.

### Claim 2 (depends on 1)

The method of claim 1 wherein the pH of the liquid bioactive coating material is greater than 8.0.

### Claim 3 (depends on 1)

The method of claim 1 wherein said protease is in solution in said liquid bioactive coating material.

### Claim 4 (depends on 1)

The method of claim 1 further comprising applying said bioactive coating material to a biological stain comprising a protein component, thereby promoting removal of said stain.

### Claim 5 (depends on 4)

The method of claim 4 wherein said biological stain is an insect stain.

### Claim 6 (depends on 4)

The method of claim 4 wherein said biological stain is present on a substrate.

### Claim 7 (depends on 6)

The method of claim 6 wherein said substrate is glass.

### Claim 8 (depends on 1)

The method of claim 1 wherein said alcohol is methanol, and said stain is an insect stain.

### Claim 9 (depends on 1)

The method of claim 1 wherein, said thermolysin-like protease is an extracellular fragment of a bacterial neutral thermolysin-like-protease from Geobacillus stearothermophilus, and wherein said stain comprises a protein.

### Claim 10

A composition for facilitating biological stain removal comprising:
a liquid bioactive coating material comprising an alcohol selected from the group consisting of methanol, ethanol, isopropyl alcohol, propanol and butanol, wherein the concentration of said alcohol in said liquid coating is from 20 to 40% wt. %, wherein the pH of the liquid coating is grater than 5.0; and
a thermolysin-like protease, wherein the protease has an activity in excess of 20,000 U/mg, said protease capable of degrading a biological stain component.

### Claim 11 (depends on 10)

The composition of claim 10 wherein said thermolysin-like protease is an active extracellular portion of bacterial neutral thermolysin-like-protease from Geobacillus stearothermophilus.

### Claim 12 (depends on 10)

The composition of claim 10 wherein the pH of said liquid bioactive coating material is greater than 8.0.

### Claim 13 (depends on 10)

The composition of claim 10 wherein said protease is in solution in said liquid bioactive coating material.

### Claim 14 (depends on 1)

The method of claim 1 wherein said alcohol is methanol, and said stain is an insect stain.

### Claim 15 (depends on 10)

The composition of claim 10 wherein said alcohol is methanol, and said stain is an insect stain.

### Claim 16 (depends on 15)

The composition of claim 15 wherein, said thermolysin-like protease is an extracellular fragment of a bacterial neutral thermolysin-like-protease from Geobacillus stearothermophilus, and wherein said liquid coating material has a pH of 8.0 or greater.

### Claim 17 (depends on 10)

The composition of claim 10, wherein said thermolysin-like protease is an extracellular fragment of a bacterial neutral thermolysin-like-protease from Geobacillus stearothermophilus, and wherein said stain comprises a protein.