# US 2021/0329941 A1 · Recombinant components and compositions for use in food products

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- Number: US 2021/0329941 A1
- Document ID: US20210329941A1
- Status: Published application
- Kind: A1
- Application number: 17/239,325
- Filed: 2021-04-23
- Published: 2021-10-28
- Earliest claimed: 2018-10-17
- Assignees: Perfect Day, Inc.
- Applicants: Perfect Day, Inc.
- Inventors: Timothy Geistlinger; Heather Jensen; Ravirajsinh Jhala; Hendrik Meerman; Balakrishnan Ramesh; Ty Wagoner; Timothy Scott Johnson; Vincent Wei-Xiang Wu; Francesca Manea
- Web page: https://patentgrep.com/patent/US20210329941A1

## CPC

- A23J 1/202: {Casein or caseinates}
- A23G 9/38: containing peptides or proteins (characterised by the dairy products used A23G9/40)
- A23J 1/006: {from vegetable materials (A23J1/005, A23J1/12 and A23J1/14 take precedence)}
- A23J 1/008: {from microorganisms (A23J1/18 takes precedence)}
- A23J 1/009: {from unicellular algae (seaweed A23J1/006)}
- A23J 1/20: from milk, e.g. casein (curds or cheese A23C); from whey
- A23J 1/205: {from whey, e.g. lactalbumine}
- A23L 29/00: Foods or foodstuffs containing additives (containing additives for modifying the nutritive qualities A23L33/10; containing substantially indigestive additives, e.g. dietary fibres, A23L33/21; containing additives for preservation A23B); Preparation or treatment thereof
- A23L 29/06: {Enzymes}
- C07K 14/4717: {Plasma globulins, lactoglobulin}
- C12N 1/14: Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor
- C12N 9/16: acting on ester bonds (3.1)
- C12N 9/18: Carboxylic ester hydrolases {(3.1.1)}
- C12N 9/20: Triglyceride splitting, e.g. by means of lipase
- C12P 21/02: having a known sequence of two or more amino acids, e.g. glutathione
- C12Y 301/01074: Cutinase (3.1.1.74)
- A23V 2002/00: Food compositions, function of food ingredients or processes for food or foodstuffs
- C12R 2001/885: Trichoderma
- C12Y 301/01001: Carboxylesterase (3.1.1.1)
- C12Y 301/01003: Triacylglycerol lipase (3.1.1.3)
- C12Y 301/01004: Phospholipase A2 (3.1.1.4)
- C12Y 301/01005: Lysophospholipase (3.1.1.5)
- C12Y 301/01006: Acetylesterase (3.1.1.6)
- C12Y 301/04003: Phospholipase C (3.1.4.3)
- A23J: PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23G: COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23L: FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF (preservation thereof A23B)
- C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00)
- 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)}
- C12Y: ENZYMES
- A23V: INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- C12R: INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS

## Abstract

Provided are methods for producing food products comprising recombinant components, and compositions used in and food products produced by such methods.

## Claims

### Claim at position 1

1-135. (canceled)

### Claim 136

A method for producing a food product comprising a recombinant component produced by a recombinant microbial host cell capable of producing the recombinant component, wherein the recombinant microbial host cell comprises a genetic modification that decreases or essentially eliminates activity or expression of an esterase compared to activity or expression of the esterase comprised in a corresponding recombinant microbial host cell, and wherein the esterase comprises a cutinase.

### Claim 137 (depends on 136)

The method of claim 136, wherein the esterase consists of a cutinase.

### Claim 138 (depends on 137)

The method of claim 137, wherein the cutinase comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0RH85, A2QBP1, A0A2T4BJB5, A5ABE6, A2R2W3, G0R6X2, and G2QH51.

### Claim 139 (depends on 138)

The method of claim 138, wherein the cutinase comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequence G0RH85.

### Claim 140 (depends on 136)

The method of claim 136, wherein the esterase further comprises a carboxylic ester hydrolase.

### Claim 141 (depends on 136)

The method of claim 136, wherein the esterase further comprises a triacylglycerol lipase.

### Claim 142 (depends on 136)

The method of claim 136, wherein the esterase further comprises a phospholipase A2.

### Claim 143 (depends on 136)

The method of claim 136, wherein the esterase further comprises a lysophospholipase.

### Claim 144 (depends on 136)

The method of claim 136, wherein the esterase further comprises an acetylesterase.

### Claim 145 (depends on 136)

The method of claim 136, wherein the esterase further comprises a phospholipase C.

### Claim 146 (depends on 136)

The method of claim 136, wherein the esterase further comprises a GDSL lipase.

### Claim 147 (depends on 136)

The method of claim 136, wherein the esterase further comprises an esterase that comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0RIU1, A2R1N7, G0RBM4, A0A2T4BBP9, G2Q379, A0A2T4AZ21, A2R8Z3, A2QPC2, G0RCG3, A0A2T4BCL7, A0A2T4B416, and A0A2T4BNI9.

### Claim 148 (depends on 136)

The method of claim 136, wherein the esterase further comprises an esterase that comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0R9X3 and G0RHJ4.

### Claim 149 (depends on 136)

The method of claim 136, wherein the esterase further comprises an esterase that comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0RX90, G2Q0K1, A0A2T4B235, A2QC75, G0RLB0, G0RLB7, G0RWS1, and A2QF42.

### Claim 150 (depends on 136)

The method of claim 136, wherein the esterase further comprises an esterase that comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0RJ76, G0RKL4, and G0RRK3.

### Claim 151 (depends on 136)

The method of claim 136, wherein the esterase further comprises an esterase that comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0REM9, A0A2T4BFY3, A2QAD7, G0RGQ7, G0RJC6, G0RQNS, G0RTR6, G0RBZ6, and G0RDK5.

### Claim 152 (depends on 136)

The method of claim 136, wherein the esterase further comprises an esterase that comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0R707, A2QM14, and A2R709.

### Claim 153 (depends on 136)

The method of claim 136, wherein the esterase further comprises an esterase that comprises an amino acid sequence that is at least 80% identical to a sequence selected from UniProt sequences G0RBJ0, G0RJY0, and G0RMI3.

### Claim 154 (depends on 136)

The method of claim 136, wherein the recombinant microbial host cell is selected from the group consisting of a recombinant bacterial host cell, a recombinant yeast host cell, and a recombinant filamentous fungal host cell.

### Claim 155 (depends on 154)

The method of claim 154, wherein the recombinant filamentous fungal host cell is derived from a filamentous fungal host cell selected from the group consisting of an Aspergillus niger host cell, a Myceliophthora thermophila host cell, a Trichoderma reesei host cell, and a Trichoderma citrinoviride host cell.

### Claim 156 (depends on 136)

The method of claim 136, wherein the recombinant component is a recombinant protein.

### Claim 157 (depends on 156)

The method of claim 156, wherein the recombinant protein is a recombinant milk protein.

### Claim 158 (depends on 156)

The method of claim 156, wherein the recombinant milk protein is selected from the group consisting of recombinant α-lactalbumin, recombinant β-lactoglobulin, recombinant lactoferrin, recombinant transferrin, recombinant serum albumin, recombinant lactoperoxidase, recombinant glycomacropeptide, recombinant β-casein, recombinant γ-casein, recombinant κ-casein, recombinant α-S1-casein, and recombinant α-S2-casein.

### Claim 159 (depends on 136)

The method of claim 136, wherein the method further comprises the step of culturing the recombinant microbial host cell under a condition suitable for further decreasing or essentially eliminating the activity of the esterase.

### Claim 160 (depends on 136)

The method of claim 136, wherein the method further comprises the step of purifying the recombinant component away from the esterase to obtain a preparation comprising the recombinant component and a decreased or essentially eliminated activity of the esterase compared to a corresponding preparation.

### Claim 161 (depends on 160)

The method of claim 160, wherein the preparation is a powder.

### Claim 162 (depends on 136)

The food product of claim 136, wherein the food product is a dairy product substitute.

### Claim 163 (depends on 162)

The food product of claim 162, wherein the dairy product substitute is a substitute of a dairy product selected from the group consisting of milk, yogurt, cheese, dairy spreads, cream, frozen confections, butter, and dairy powders.