# US 2021/0276907 A1 · Method for extracting antibacterial peptides and albumin from pea whey wastewater

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 2021/0276907 A1
- Document ID: US20210276907A1
- Status: Published application
- Kind: A1
- Application number: 17/169,542
- Filed: 2021-02-07
- Published: 2021-09-09
- Earliest claimed: 2020-03-09
- Assignees: Yantai Shuangta Food Co., Ltd.
- Applicants: Yantai Shuangta Food Co., Ltd.
- Inventors: Shucheng Zhang; Jinjie Yang; Qingjia Zang; Shimin Wu; Guodong Yang
- Web page: https://patentgrep.com/patent/US20210276907A1

## CPC

- C02F 9/00: Multistage treatment of water, waste water or sewage
- A23J 1/16: from waste water of starch-manufacturing plant or like wastes
- A23J 3/14: Vegetable proteins
- A23J 3/346: {of vegetable proteins}
- B01D 1/26: Multiple-effect evaporating
- B01D 61/027: {Nanofiltration}
- B01D 61/0271: {comprising multiple nanofiltration steps}
- B01D 61/145: {Ultrafiltration}
- B01D 61/147: {Microfiltration}
- B01D 61/149: {Multistep processes comprising different kinds of membrane processes selected from ultrafiltration or microfiltration}
- B01D 61/58: Multistep processes
- B01D 69/02: characterised by their properties
- C07K 1/36: by a combination of two or more processes of different types
- C07K 14/415: from plants
- B01D 2311/2649: Filtration
- B01D 2311/2673: Evaporation
- B01D 2311/2676: Centrifugal separation
- B01D 2311/2692: Sterilization
- B01D 2325/0283: Pore size
- B01D 2325/20: Specific permeability or cut-off range
- C02F 1/001: {Processes for the treatment of water whereby the filtration technique is of importance (C02F1/44 takes precedence; construction of filters in general B01D24/00 - B01D41/00)}
- C02F 1/06: Flash evaporation
- C02F 1/38: by centrifugal separation
- C02F 1/442: {by nanofiltration}
- C02F 1/444: {by ultrafiltration or microfiltration}
- C02F 1/66: by neutralisation; pH adjustment (for degassing C02F1/20; using ion-exchange C02F1/42; for flocculation or precipitation of suspended impurities C02F1/52; for removing dissolved compounds C02F1/58)
- C02F 2101/38: containing nitrogen
- C02F 2103/32: from the food or foodstuff industry, e.g. brewery waste waters
- C02F 2301/08: Multistage treatments, e.g. repetition of the same process step under different conditions
- C02F: TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE (separation in general B01D; special arrangements on waterborne vessels of installations for treating water, waste water or sewage, e.g. for producing fresh water, B63J; adding materials to water to prevent corrosion C23F; treating radioactively-contaminated liquids G21F9/04; regeneration of reactants for recirculation into processes, see the relevant places for the processes)
- A23J: PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- B01D: SEPARATION (separating solids from solids by wet methods B03B, B03D; by pneumatic jigs or tables B03B; by other dry methods B07; magnetic or electrostatic separation of solid materials from solid materials or fluids, separation by high-voltage electric fields B03C; centrifuges, vortex apparatus B04; presses per se for squeezing-out liquid from liquid-containing material B30B9/02; treatment of water C02F, e.g. softening by ion-exchange C02F1/42; {arrangements of air intake cleaners in gas turbine plants F02C7/05} ; arrangements or mounting of filters in air-conditioning, air-humidification or ventilation F24F13/28)
- 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 disclosure provides a method for extracting antibacterial peptides and albumin from pea whey wastewater, which includes following steps: in extracting the albumin, centrifuging with the pea whey wastewater generated during pea protein processing as raw material; controlling temperature and exchanging heat to adjust temperature of the raw material; sequentially performing a microfiltrating, nanofiltration, ultrafiltration and secondary nanofiltration to obtain an albumin slurry; performing a multi-effect concentration on the albumin slurry; adding an alkaline substances to adjust pH; sterilizing and drying to finally obtain the albumin, which realizes a targeted extraction of the albumin with small molecular weight in the pea whey wastewater and avoids resource waste. The albumin with small molecular weight and the antibacterial peptides in the pea whey water are effectively recycled, thus avoiding environmental pollution caused by improper treatment of the pea whey water and realizing resource reuse.

## Claims

### Claim 1

A method for extracting antibacterial peptides from pea whey wastewater, comprising following steps: centrifuging with the pea whey wastewater generated during pea protein processing as raw material; controlling temperature and exchanging heat to adjust temperature of the raw material; sequentially performing a microfiltrating, nanofiltration, ultrafiltration and secondary nanofiltration to obtain an albumin slurry; performing a multi-effect concentrating on the albumin slurry; adding an alkaline substances to adjust pH; sterilizing and drying to obtain the albumin;
adding water to the albumin to obtain another albumin slurry, pretreating in water bath, cooling and stirring on a constant temperature magnetic stirrer, adding papain and dipping alkali solution to keep pH of enzymatic hydrolysate constant; boiling to inactivate enzyme after full enzymolysis and cooling, adjusting the pH to 7.0; centrifuging and concentrating supernatant by rotary evaporation, and sterilizing with a 0.22 μm syringe filter to obtain primary product of the antibacterial peptides; and
passing the primary product of the antibacterial peptides through an extractor, which is cleaned and activated in advance with methanol aqueous solution and then cleaned and balanced by aqueous trifluoroacetic acid solution; sequentially passing the methanol aqueous solution and the aqueous trifluoroacetic acid solution through the extractor and collecting penetrating liquid, vacuum concentrating the penetrating liquid by a rotary evaporator to get concentrated antibacterial peptide solution, taking out the concentrated antibacterial peptide solution and freezing at −15° C., and drying the frozen antibacterial peptides in a vacuum freeze dryer to obtain purified antibacterial peptides;
wherein PH value of the antibacterial peptides is 2.56 to 2.78, as measured by a pH meter; and
an antibacterial peptide molecule contains glycine, cysteine, arginine, lysine, histidine, alanine, threonine, aspartic acid, leucine, phenylalanine, serine, glutamic acid, valine, methionine, or tyrosine.

### Claim 2

A method for extracting albumin from pea whey wastewater, comprising following steps:
centrifuging with the pea whey wastewater generated during pea protein processing as raw material; controlling temperature and exchanging heat to adjust temperature of the raw material;
sequentially performing a microfiltrating, nanofiltration, ultrafiltration and secondary nanofiltration to obtain an albumin slurry; performing a multi-effect concentration on the albumin slurry; adding an alkaline substances to adjust pH; and sterilizing and drying to obtain the albumin.

### Claim 3 (depends on 2)

The method for extracting the albumin from the pea whey wastewater according to claim 2, specifically comprising:
1) the centrifuging: centrifuging with the pea whey wastewater as the raw material;
2) the controlling temperature and exchanging heat: performing a heat exchange on the centrifuged pea whey wastewater;
3) the performing the microfiltration: performing the microfiltration on the pea whey wastewater after the heat exchange;
4) the performing the nanofiltration: removing 93% to 95% of water in the pea whey wastewater after the microfiltration by using a nanofiltration module and adjusting system pressure and filtration temperature;
5) the performing the ultrafiltration: eluting and separating for 5 to 10 times with an ultrafiltration membrane to obtain a crude albumin slurry;
6) the performing the secondary nanofiltration: washing the crude albumin with clean water added and using an anti-pollution nanofiltration membrane, and removing the clean water for washing after the washing is complete;
7) the performing the multi-effect concentration: adding the washed albumin slurry into an evaporator for evaporation and concentration;
8) the adjusting the pH: adding the alkaline substance into a stainless steel tank and adjusting the pH value; and
9) sterilizing and drying.

### Claim 4 (depends on 2)

The method for extracting the albumin from the pea whey wastewater according to claim 2, wherein the temperature in controlling temperature and exchanging heat is 40 to 50° C.

### Claim 5 (depends on 2)

The method for extracting the albumin from the pea whey wastewater according to claim 2, wherein a membrane used in the microfiltration is a silicon carbide membrane or a ceramic membrane.

### Claim 6 (depends on 5)

The method for extracting the albumin from the pea whey wastewater according to claim 5, wherein a pore size of the silicon carbide film is 10 nanometers to 30 micrometers.

### Claim 7 (depends on 2)

The method for extracting the albumin from the pea whey wastewater according to claim 2, wherein during the nanofiltration, the system pressure is adjusted to 18 to 25 bar, and the filtration temperature is 40 to 65° C.

### Claim 8 (depends on 2)

The method for extracting the albumin from the pea whey wastewater according to claim 2, wherein a pore size of an ultrafiltration membrane is configured to provide a molecular weight cut-off of 1000 to 5000 Daltons.

### Claim 9 (depends on 3)

The method for extracting the albumin from the pea whey wastewater according to claim 3, wherein during the secondary nanofiltration, the clean water for washing is the water of 93% to 95% obtained in the nanofiltration in step 4).

### Claim 10 (depends on 2)

The method for extracting the albumin from the pea whey wastewater according to claim 2, wherein during the multi-effect concentration, a vapor pressure is 0.6-0.8 MPa, and a concentration of the concentrated albumin slurry is 25% to 50%; and during the adjusting the pH, the alkaline substance is added at a temperature of 40 to 65° C. to adjust the pH to be 6.5 to 8.

### Claim 11 (depends on 3)

The method for extracting the albumin from the pea whey wastewater according to claim 3, wherein the temperature in controlling temperature and exchanging heat is 40 to 50° C.

### Claim 12 (depends on 3)

The method for extracting the albumin from the pea whey wastewater according to claim 3, wherein a membrane used in the microfiltration is a silicon carbide membrane or a ceramic membrane.

### Claim 13 (depends on 3)

The method for extracting the albumin from the pea whey wastewater according to claim 3, wherein during the nanofiltration, the system pressure is adjusted to 18 to 25 bar, and the filtration temperature is 40 to 65° C.

### Claim 14 (depends on 3)

The method for extracting the albumin from the pea whey wastewater according to claim 3, wherein a pore size of an ultrafiltration membrane is configured to provide a molecular weight cut-off of 1000 to 5000 Daltons.

### Claim 15 (depends on 3)

The method for extracting the albumin from the pea whey wastewater according to claim 3, wherein during the multi-effect concentration, a vapor pressure is 0.6-0.8 MPa, and a concentration of the concentrated albumin slurry is 25% to 50%; and during the adjusting the pH, the alkaline substance is added at a temperature of 40 to 65° C. to adjust the pH to be 6.5 to 8.