# US 12,004,537 B2 · Compositions comprising fabaceae family plant components, processes of preparation and uses thereof

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- Number: US 12,004,537 B2
- Document ID: US12004537B2
- Status: Granted
- Kind: B2
- Application number: 18/033,134
- Filed: 2021-10-21
- Published: 2024-01-04
- Granted: 2024-06-11
- Earliest claimed: 2020-10-21
- Assignees: Virentia Innovation Inc.
- Applicants: Virentia Innovation Inc.
- Inventors: Pierre Lavallée; Réjean Desgagnés; Laurence Cambron-Fortin; Louis-Philippe Vézina; Pierre Talbot
- Web page: https://patentgrep.com/patent/US12004537B2

## CPC

- A23K 10/30: from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms (obtained by microbiological or biochemical processes, e.g. using yeasts or enzymes, A23K10/10)
- C12Y 301/03011: Fructose-bisphosphatase (3.1.3.11)
- A01H 6/544: {Medicago sativa [alfalfa]}
- A23K 20/147: Polymeric derivatives, e.g. peptides or proteins
- A23K 20/158: Fatty acids; Fats; Products containing oils or fats
- A23K 20/174: Vitamins
- A23K 20/179: Colouring agents, e.g. pigmenting or dyeing agents
- A23K 50/80: for aquatic animals, e.g. fish, crustaceans or molluscs
- A23L 33/105: Plant extracts, their artificial duplicates or their derivatives
- A23L 33/115: Fatty acids or derivatives thereof; Fats or oils
- A23L 33/12: Fatty acids or derivatives thereof
- A23L 33/15: Vitamins
- A23L 33/185: Vegetable proteins
- C07K 14/415: from plants
- C12Y 401/01039: Ribulose-bisphosphate carboxylase (4.1.1.39)
- A23K 20/163: Sugars; Polysaccharides
- A23L 33/21: Addition of substantially indigestible substances, e.g. dietary fibres
- A23K: FODDER
- C12Y: ENZYMES
- A01H: NEW PLANTS OR {NON-TRANSGENIC} PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- 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)

## Abstract

The present disclosure relates to processes for recovering valuable products from Fabaceae family plant fractions, in particular from Medicago sativa ssp. The processes disclosed herein include processes for obtaining macrofibers, microfibers, a saponin precursor, chloroplast liquid and dry compositions and a Rubisco precursor. There is also disclosed herein processes for extracting from Fabaceae family plants valuable compounds such as proteins, enzymes, peptides, amino acids, fatty acids, fatty alcohols, terpenes, phenols and pigments. The processes may comprise at least one of separating plant fibers while attenuating shear forces, maintaining the temperature at or below 45° C., maintaining the pH above 4 and adding antioxidant and/or antimicrobial agents. Compositions comprising these recovered Fabaceae family plant products and uses thereof are also disclosed.

## Claims

### Claim 1

A dry chloroplast composition comprising chloroplasts isolated from Fabaceae family plants, wherein at least 75% of the chloroplasts comprised in the composition are intact as determined by dynamic light scattering measurement and analysis,
and wherein said dry chloroplast composition has at least one ratio chosen from:
a ratio of chlorophyll/fructose 1,6-biphosphatase (FBPP) of about 50 to about 130, about 80 to about 120, about 90 to about 110, about 95 to about 105, about 97 to about 100 or about 98 to 100 the ratio of chlorophyll/FBPP being
chlorop
⁢
hyll
⁢
in
⁢
(
mg
/
g
)
FBPP
⁢
in
⁢
(
intensity
/
mg
⁢
powder
)
×
10000000
,
the FBPP intensity being measured by immunoblot,
a ratio of Rubisco/chlorophyll of about 25 to about 65, about 30 to about 60, about 35 to about 55, about 40 to about 50, or about 43 to about 49, the ratio of Rubisco/chlorophyll being
(
Rubisco
⁢
in
⁢
intensity
chlorophyll
⁢
in
⁢
(
mg
/
g
)
)
/
1000
,
wherein the Rubisco intensity being measured by immunoblot,
a ratio of Rubisco/chlorophyll of about 2.5 ng/mg to about 6.5 ng/mg, about 3.0 ng/mg to about 6.0 ng/mg, about 3.5 ng/mg to about 5.5 ng/mg, about 4.0 ng/mg to about 5.0 ng/mg, or about 4.3 ng/mg to about 4.7 ng/mg, the ratio of Rubisco/chlorophyll being
(
Rubisco
⁢
in
⁢
ng
/
mg
⁢
powder
chlorophyll
⁢
in
⁢
(
mg
/
g
)
)
/
100
⁢
0
.
and
a ratio of Rubisco/FBPP of about 18 to about 100, about 40 to about 90, about 50 to about 80, about 55 to about 75, or about 60 to about 70, the ratio of Rubisco/chlorophyll being
(
Rubisco
⁢
in
⁢
ng
/
mg
⁢
powder
FBPP
⁢
in
⁢
intensity
/
mg
)
×
100
;
and
wherein the protein content of the composition is greater than 45 wt. %.

### Claim 2 (depends on 1)

The composition of claim 1, wherein the plant is alfalfa.

### Claim 3 (depends on 1)

The composition of claim 1, having a moisture content of less than 4%, less than 3%, less than 2% or less than 1%.

### Claim 4 (depends on 1)

The composition of claim 1, wherein the composition is in dry form.

### Claim 5 (depends on 1)

The composition of claim 1, wherein the composition is in powder form.

### Claim 6 (depends on 1)

The composition of claim 1, further comprising an antioxidant, an antimicrobial agent, optionally a bacteriostatic or a bactericide, a fungicide, or mixtures thereof.

### Claim 7 (depends on 1)

The composition of claim 1, further comprising an antioxidant and/or an antimicrobial agent.

### Claim 8 (depends on 7)

The composition of claim 7, wherein the antioxidant and/or the antimicrobial agent is citric acid, metabisulfite, benzoate, optionally sodium metabisulfite, potassium metabisulfite, sodium benzoate or potassium benzoate.

### Claim 9 (depends on 1)

The composition of claim 1, further comprising omega-3 fatty acids, omega-6 fatty acids, vitamins, or mixtures thereof.

### Claim 10 (depends on 1)

The composition of claim 1, wherein about 40 wt. % to about 90 wt. %, about 50 wt. % to about 90 wt. %, about 60 wt. % to about 90 wt. %, about 70 wt. % to about 90 wt. % or about 75 wt. % to about 90 wt. % of the solid content consists of chloroplasts.

### Claim 11 (depends on 1)

The composition of claim 1, wherein at least 50%, 60%, 70%, 80% or 90% of solid particles comprised in the composition have an average size of about 5 microns to about 10 microns.

### Claim 12 (depends on 1)

The composition of claim 1, wherein at least 50%, 60%, 70%, 80% or 90% of the chloroplasts have a preserved outer membrane integrity.

### Claim 13 (depends on 1)

The composition of claim 1, wherein at least 50%, 60%, 70%, 80% or 90% of the chloroplasts have a preserved inner membrane integrity.

### Claim 14 (depends on 1)

The composition of claim 1, wherein at least 50%, 60%, 70%, 80% or 90% of the chloroplasts have maintained metabolic activity as compared to chloroplasts comprised in a reference Fabaceae family plant.

### Claim 15 (depends on 1)

The composition of claim 1, wherein at least 50%, at least 60%, at least 70% or at least 80% of the chloroplasts are intact chloroplasts as determined by dynamic light scattering measurement and analysis.

### Claim 16 (depends on 1)

The composition of claim 1, having a protein content of about 45 wt. % to about 55 wt. % or about 48 wt. % to about 55 wt. %.

### Claim 17 (depends on 1)

The composition of claim 1, having a protein content of about 48 wt. % to about 52 wt. %.

### Claim 18 (depends on 1)

The composition of claim 1, having a beta-carotene content of greater than 300,000 IU/100 g.

### Claim 19 (depends on 1)

The composition of claim 1, having a chlorophyll content of greater than 20 mg/g, greater than 25 mg/g or greater than 30 mg/g.

### Claim 20 (depends on 1)

The composition of claim 1, having a xanthophyll lutein content of greater than 1.6 mg/g.

### Claim 21 (depends on 1)

The composition of claim 1, having an antioxidant content of greater than 20,000 μmole Trolox equivalents/100 g.

### Claim 22 (depends on 1)

The composition of claim 1, having an antioxidant content of about 20,000 000 μmole Trolox equivalents/100 g to about 24,000 000 μmole Trolox equivalents/100 g.

### Claim 23 (depends on 1)

The composition of claim 1, having a lipid content of about 5 wt. % to about 15 wt. %, about 10 wt. % to about 15 wt. % or about 10 wt. % to about 12 wt. %.

### Claim 24 (depends on 1)

The composition of claim 1, having an omega-3 fatty acid content, optionally an eicosapentaenoic acid and/or a docosahexaenoic acid content, of greater than 2 wt. %.

### Claim 25 (depends on 1)

The composition of claim 1, having an omega-3 fatty acid content of about 2 wt. % to about 10 wt. % or about 2 wt. % to about 6 wt. %.

### Claim 26 (depends on 1)

The composition of claim 1, having an omega-6 fatty acid content of greater than 1%.

### Claim 27 (depends on 1)

The composition of claim 1, having an omega-6 fatty acid content of about 1% to about 10% or about 1% to about 4%.

### Claim 28 (depends on 1)

The composition of claim 1, having an omega-3 fatty acid/omega-6 fatty acid ratio of about 1.5 to about 3.

### Claim 29 (depends on 1)

The composition of claim 1, having a lipid/protein ratio of about 0.2 to about 0.4.

### Claim 30 (depends on 1)

The composition of claim 1, having a pH of less than 5.0, less than 4.8, less than 4.7, less than 4.6, less than 4.5, less than 4.4, less than 4.3 or less than 4.2.

### Claim 31 (depends on 1)

The composition of claim 1, having a protein content greater than 50 wt. % and a chlorophyll content greater than 25 mg/g, wherein at least 70% of the chloroplasts are intact chloroplasts as determined by dynamic light scattering measurement and analysis.

### Claim 32 (depends on 1)

The composition of claim 1, wherein the composition is encapsulated in a capsule, optionally an opaque capsule.