# US 2025/0154491 A1 · Process of obtaining a protein preparation

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- Number: US 2025/0154491 A1
- Document ID: US20250154491A1
- Status: Published application
- Kind: A1
- Application number: 18/837,593
- Filed: 2023-02-09
- Published: 2025-05-15
- Earliest claimed: 2022-02-10
- Assignees: Instituto Superior De Agronomia [Pt/Pt]
- Applicants: Instituto Superior De Agronomia
- Inventors: Ricardo Manuel DE SEIXAS BOAVIDA FERREIRA; Madalena DE MATOS ÁGUAS GRÁCIO; Giovanni DEL FRARI
- Web page: https://patentgrep.com/patent/US20250154491A1

## CPC

- C12N 9/88: Lyases (4.)
- A23J 1/001: {from waste materials, e.g. kitchen waste}
- A23J 1/006: {from vegetable materials (A23J1/005, A23J1/12 and A23J1/14 take precedence)}
- A23J 1/007: {from leafy vegetables, e.g. alfalfa, clover, grass}
- A23J 1/008: {from microorganisms (A23J1/18 takes precedence)}
- A23J 1/009: {from unicellular algae (seaweed A23J1/006)}
- A23J 1/14: from leguminous or other vegetable seeds; from press-cake or oil-bearing seeds
- A23J 3/14: Vegetable proteins
- A23J 3/20: Proteins from microorganisms or unicellular algae
- C12Y 401/01039: Ribulose-bisphosphate carboxylase (4.1.1.39)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A23J: PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- C12Y: ENZYMES

## Abstract

A process of obtaining a protein preparation from biological tissues selected from the group comprising seeds eg. legume seeds, algae eg. macro or microalgae, bacteria eg. cyanobacteria, animal products, said process comprises the steps of: lysing said biological tissues to extract protein; and submitting at least a portion of said extracted material to an activated charcoal; whereby a portion is removed by or adsorbed onto said activated charcoal; and/or submitting a portion of extracted material to a solution containing EDTA eg. of 10 to 40 mM; and/or submitting a portion of extracted material to a solution containing NaCl eg. of 5 to 15% (w/v); and/or submitting a portion of extracted material to a bentonite concentration; and/or submitting a portion of extracted material to a solution containing phytate eg. of 7.5 to 30 mM; and/or submitting a portion of extracted material to a phosphate eg. of 25 to 100 mM; whereby one or more of the following proteins are isolated: legume seed proteins, RuBisCO, soluble protein from leafy vegetables either edible or non-edible, macro or micro algae protein, cyanobacteria protein, animal products protein.

## Claims

### Claim 1

A process of obtaining a RuBisCO preparation from photosynthetic material, comprising the steps of:
lysing said photosynthetic material to extract RuBisCO accompanied by a fraction of the chlorophylls of said photosynthetic material; and
submitting at least a portion of said extracted material to an activated charcoal; whereby a further portion is removed by or adsorbed onto said activated charcoal.

### Claim 2 (depends on 1)

The process of claim 1, wherein said further portion comprises chlorophyll pigments.

### Claim 3 (depends on 1)

The process of claim 1, comprising a step of grinding leaves containing photosynthetic material and adding water to obtain an aqueous solution.

### Claim 4 (depends on 3)

The process of claim 3, comprising the step of agitating and/or centrifuging said aqueous solution in order to obtain a pellet of insoluble particles and a supernatant.

### Claim 5 (depends on 4)

The process of claim 4, comprising the step of adding said activated charcoal to said supernatant and further processing the solution whereby said activated charcoal adsorbs chlorophyll pigments.

### Claim 6

(canceled)

### Claim 7 (depends on 5)

The process of claim 5, wherein said further processing results in a pellet of low molecular mass molecules, ions and/or atoms and a supernatant of soluble proteins which were non-adsorbed onto said activated charcoal.

### Claim 8 (depends on 1)

The process of claim 1, wherein the process further comprises a step of centrifugation to separate out pellets and a supernatant comprising the soluble proteins which were not adsorbed onto the activated charcoal.

### Claim 9 (depends on 1)

The process of claim 1, which further comprises one or more further steps of nanofiltration, ultrafiltration or microfiltration; whereby soluble RuBisCO is isolated.

### Claim 10 (depends on 9)

The process according to claim 9, wherein said nano-filtration employs a PES (polyethersulfone) membrane of for example 0.22 μm.

### Claim 11 (depends on 9)

The process according to claim 9, wherein said nano-filtration employs with a molecular weight cut-off membrane of 100 kDa or less.

### Claim 12

(canceled)

### Claim 13 (depends on 1)

A RubisCO preparation obtained by a process according to claim 1.

### Claim 14

A process of obtaining a protein preparation from biological tissues selected from the group comprising seeds, algae, cyanobacteria and animal products, said process comprising the steps of:
lysing said biological tissues to extract protein; and
submitting at least a portion of said extracted material to an activated charcoal; whereby a portion is removed by or adsorbed onto said activated charcoal; and/or submitting a portion of extracted material to a solution containing EDTA eg. of 10 to 40 mM; and/or submitting a portion of extracted material to a solution containing NaCl eg. of 2.5 to 15% (w/v); and/or submitting a portion of extracted material to a bentonite concentration; and/or submitting a portion of extracted material to a solution containing phytate eg. of 7.5 to 30 mM; and/or submitting a portion of extracted material to a phosphate eg. of 25 to 100 mM.

### Claim 15 (depends on 14)

The process of claim 14, wherein said further portion comprises chlorophyll pigments.

### Claim 16 (depends on 14)

The process of claim 14, comprising a step of grinding biological tissues and adding water to obtain an aqueous solution.

### Claim 17 (depends on 16)

The process of claim 16, comprising the step of agitating and/or centrifuging said aqueous solution in order to obtain a pellet of insoluble particles and a supernatant and further comprising the step of adding said activated charcoal to said supernatant and further processing the solution whereby said activated charcoal adsorbs a further portion.

### Claim 18

(canceled)

### Claim 19

(canceled)

### Claim 20 (depends on 17)

The process of claim 17, resulting in a pellet of low molecular mass molecules, ions and/or atoms and a supernatant of soluble proteins which were non-adsorbed onto said activated charcoal.

### Claim 21 (depends on 20)

The process of claim 20 wherein the process further comprises a step of centrifugation to separate out pellets and a supernatant comprising the soluble proteins which were not adsorbed onto the activated charcoal.

### Claim 22 (depends on 14)

The process according to claim 14, which further comprises one or more further steps of nanofiltration, ultrafiltration or microfiltration; whereby one or more of the following proteins are isolated: legume seed proteins, RuBisCO, soluble protein from leafy vegetables either edible or non-edible, macro or micro algae protein, cyanobacteria protein, animal products protein.

### Claim 23 (depends on 22)

The process according to claim 22, wherein said nano-filtration employs a PES (polyethersulfone) membrane of for example 0.22 μm.

### Claim 24 (depends on 22)

The process according to claim 22, wherein said nano-filtration employs an ultra-filtration membrane with a molecular weight cut-off membrane of 100 kDa or less.

### Claim 25

(canceled)

### Claim 26

(canceled)