# US 2021/0277424 A1 · Bioremediation of petrochemical-containing substrates using fungi

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- Number: US 2021/0277424 A1
- Document ID: US20210277424A1
- Status: Published application
- Kind: A1
- Application number: 17/274,425
- Filed: 2019-09-09
- Published: 2021-09-09
- Earliest claimed: 2018-09-11
- Assignees: Mycocycle, LLC
- Applicants: Mycocycle, LLC
- Inventors: JOANNE RODRIGUEZ; PETER MCCOY
- Web page: https://patentgrep.com/patent/US20210277424A1

## CPC

- B09C 1/00: Reclamation of contaminated soil
- C12P 1/02: by using fungi
- B01D 53/85: with gas-solid contact
- C02F 3/347: {Use of yeasts or fungi (C02F3/322 takes precedence)}
- C12N 1/14: Fungi (culture of mushrooms A01G18/00; as new plants A01H15/00); Culture media therefor
- B01D 2251/95: Specific microorganisms
- C02F 2101/20: Heavy metals or heavy metal compounds
- C02F 2101/327: {Polyaromatic Hydrocarbons [PAH's]}
- C02F 2101/34: containing oxygen
- C02F 2305/06: Nutrients for stimulating the growth of microorganisms
- C12N 2500/76: from plants
- B09C: RECLAMATION OF CONTAMINATED SOIL (gatherers for removing stone or the like from the soil A01B43/00; sterilising soil by steam A01G11/00; removing undesirable matter, e.g. rubbish, from the land E01H15/00)
- 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)}
- 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)
- 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)
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

The present disclosure provides methods and compositions for bioremediating solid petrochemical-containing scrap into biomass.

## Claims

### Claim at position 1

1-53. (canceled)

### Claim 54

A method of bioremediating solid petrochemical-containing scrap material, the method comprising the steps of:
a. breaking down the scrap material into discrete pieces;
b. mixing the broken-down scrap pieces with a growth medium selected from the group consisting of sawdust, paper, hemp, straw, gypsum and cardboard to form a scrap-growth medium mixture;
c. sterilizing the scrap-growth medium mixture;
d. homogenizing the scrap-growth medium mixture;
e. hydrating the scrap-growth medium mixture to a moisture content of 50-75%;
f. inoculating the hydrated scrap-growth medium mixture with a saprotrophic fungus species selected from the group consisting of Agrocybe spp., Amanita spp., Armillaria spp., Auricularia spp., Cerrena spp., Coprinus spp., Cyathus spp., Daedalea spp., Daedaleopsis spp., Daldinia spp., Echinodontium spp., Exidia spp., Fistulina spp., Flammulina spp., Fomes spp., Grifola spp., Hericium spp., Heterobasidion spp., Hypsizygus spp., Inonotus spp., Lenzites spp., Marasmius spp., Phanerochaete spp., Pisolithus spp., Sparassis spp., Strobilomyces spp., Xylaria spp., Pleurotus spp., Ganoderma spp., Trametes spp. Schizophyllum spp., Irpex spp. and Lentinula spp;
g. exposing the hydrated scrap-growth medium-fungus mixture to air; and
h. incubating the scrap-growth medium-water-fungus mixture.

### Claim 55 (depends on 54)

The method of claim 54, wherein the petrochemical in the scrap comprises asphalt.

### Claim 56 (depends on 54)

The method of claim 54, wherein the petrochemical in the scrap comprises a polycyclic aromatic hydrocarbon.

### Claim 57 (depends on 56)

The method of claim 56, wherein the polycyclic aromatic hydrocarbon is selected from the group consisting of pyrene, naphthalene, and anthracene.

### Claim 58 (depends on 54)

The method of claim 54, wherein the petrochemical in the scrap comprises a plastic.

### Claim 59 (depends on 58)

The method of claim 58, wherein the plastic is selected from the group consisting of polyethylene, polypropylene, ethylene acrylate copolymer, ethyl-vinyl-acetate (EVA), polyvinyl chloride (PVC), ethylene propylene, ethylene propylene diene monomer rubber, and polyolefin.

### Claim 60 (depends on 54)

The method of claim 54, wherein the sterilizing step is accomplished chemically.

### Claim 61 (depends on 60)

The method of claim 60, wherein the chemical sterilizing step is accomplished with bleach or hydrogen peroxide.

### Claim 62 (depends on 54)

The method of claim 54, wherein the sterilizing step is accomplished by heat.

### Claim 63 (depends on 54)

The method of claim 54, wherein the sterilizing step is accomplished by autoclave.

### Claim 64

A bioremediating composition, comprising:
a. A plurality of discrete pieces of solid petrochemical-containing scrap material;
b. growth medium selected from the group consisting of sawdust, paper, hemp, straw, gypsum and cardboard;
c. air;
d. water;
e. a sterilizing agent; and
f. a fungal culture selected from the group consisting of: Agrocybe spp., Amanita spp., Armillaria spp., Auricularia spp., Cerrena spp., Coprinus spp., Cyathus spp., Daedalea spp., Daedaleopsis spp., Daldinia spp., Echinodontium spp., Exidia spp., Fistulina spp., Flammulina spp., Fomes spp., Grifola spp., Hericium spp., Heterobasidion spp., Hypsizygus spp., Inonotus spp., Lenzites spp., Marasmius spp., Phanerochaete spp., Pisolithus spp., Sparassis spp., Strobilomyces spp., Xylaria spp., Pleurotus spp., Ganoderma spp., Trametes spp. Schizophyllum spp., Irpex spp. and Lentinula spp.

### Claim 65 (depends on 64)

The composition of claim 64, wherein the petrochemical in the scrap comprises asphalt.

### Claim 66 (depends on 64)

The composition of claim 64, wherein the petrochemical in the scrap comprises a polycyclic aromatic hydrocarbon.

### Claim 67 (depends on 66)

The composition of claim 66, wherein the polycyclic aromatic hydrocarbon is selected from the group consisting of pyrene, naphthalene, and anthracene.

### Claim 68 (depends on 64)

The composition of claim 64, wherein the petrochemical in the scrap comprises a plastic.

### Claim 69 (depends on 67)

The composition of claim 67, wherein the plastic is selected from the group consisting of polyethylene, polypropylene, ethylene acrylate copolymer, ethyl-vinyl-acetate (EVA), polyvinyl chloride (PVC), ethylene propylene, ethylene propylene diene monomer rubber, and polyolefin.

### Claim 70 (depends on 64)

The composition of claim 64, wherein the sterilizing agent comprises bleach or hydrogen peroxide.