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United States Patent (10) Patent No.: US 9,572,364 B2 Langan Et Al

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USOO9572364B2 (12) United States Patent (10) Patent No.: US 9,572,364 B2 Langan et al. (45) Date of Patent: *Feb. 21, 2017 (54) METHODS FOR THE PRODUCTION AND 6,490,824 B1 12/2002 Intabon et al. USE OF MYCELIAL LIQUID TISSUE 6,558,943 B1 5/2003 Li et al. CULTURE 6,569.475 B2 5/2003 Song 9,068,171 B2 6/2015 Kelly et al. (71) Applicant: Mycotechnology, Inc., Aurora, CO 2002.01371.55 A1 9, 2002 Wasser et al. (US) 2003/0208796 Al 11/2003 Song et al. (72) Inventors: James Patrick Langan, Denver, CO 3988: A. 58: sistset al. (US); Brooks John Kelly, Denver, CO 2004f02.11721 A1 10, 2004 Stamets (US); Huntington Davis, Broomfield, 2005/0180989 A1 8/2005 Matsunaga CO (US); Bhupendra Kumar Soni, 2005/0255126 A1 11/2005 TSubaki et al. Denver, CO (US) 2005/0273875 A1 12, 2005 Elias s 2006/0014267 A1 1/2006 Cleaver et al. (73) Assignee: MYCOTECHNOLOGY, INC., Aurora, 2006/0134294 A1 6/2006 McKee et al. CO (US) 2006/0280753 A1 12, 2006 McNeary (*) Notice: Subject to any disclaimer, the term of this 2007/O160726 A1 T/2007 Fujii patent is extended or adjusted under 35 (Continued) U.S.C. 154(b) by 0 days. FOREIGN PATENT DOCUMENTS This patent is Subject to a terminal dis claimer. CN 102860541. A 1, 2013 DE 4341316 6, 1995 (21) Appl. No.: 15/144,164 (Continued) (22) Filed: May 2, 2016 OTHER PUBLICATIONS (65) Prior Publication Data Diekman "Sweeteners Facts and Fallacies: Learn the Truth About US 2016/0249660 A1 Sep. 1, 2016 the Different Types of Sweeteners to Better Counsel Patients', s Today’s Dietitian 14(9): p. 42, Sep. 2012.* JP S6291161. A machine translated by EPO website.* Related U.S. Application Data Beuchat “Indigenous Fermented Foods', in Biotechnology Set, Second Edition (eds H.-J. Rehm and G. Reed), Wiley-VCH Verlag (63) Continuation-in-part of application No. 14/836,830, GmbH. Weinheim, Germany, p. 505-559, 2001.* filed on Aug. 26, 2015. Office Action issued Feb. 19, 2014 for U.S. Appl. No. 13/844,685. Autoclave Search (2014) www.meriam-Webster.com/medlineplus/ autoclave. (60) gril appli t N. flyg. Berovic et al. (2003) “Submerged cultivation of Ganoderma s proV1S1onal app ication o. 04/423.0/. lucidum biomass and immunostimulatory effects of fungal polysac filed on Nov. 10, 2015, provisional application No. charides' J. Biotechnol. 103(1): 77-86. 62/281546, filed on Jan. 21, 2016. Bok et al. Phytochemistry (1999) “Antitumor sterols from the mycelia of Cordyceps sinensis' 51: 891-898. (51) Int. Cl. “Can You Eat Mycelium?” (2014) wiki.answers.com. A2.3L I/28 (2006.01) de Melo, Rodrigues et al. (2008) Influence of Flammulina velutipes mycelia culture conditions on antimicrobial metabolite production A2.3L. I./22 (2006.01) Mycoscience 50(1): 78-81. A2.3L I/236 (2006.01) "Eat Mycelium cakes?” (2014) Mycotopia.net. A2.3L I/23 (2006.01) E.aling mycelium (2.to itytrip Snroomery.org.St. (52) st (2006.01) "Eating Mycelium?” (2014) Zoklet.net. CPC .............. A2.3L I/22083 (2013.01); A23J 3/14 (Continued) (2013.01); A23L I/23 (2013.01); A23L I/2363 Primary Examiner — Emily Cordas (2013.01); A23L I/28 (2013.01); A23L 27/24 (74) Attorney, Agent, or Firm — Swanson & Bratschun, (2016.08); A23L 27/33 (2016.08); A23L 2786 L.L.C. (2016.08); A23L 27/88 (2016.08); A23L 31/00 (57) ABSTRACT (2016.08); A23 V 2002/00 (2013.01) The present invention includes a method for enhancing the (58) Field of Classification Search taste of a food product, which includes the steps of culturing None a submerged mycelial liquid tissue culture in a media, See application file for complete search history. collecting the Supernatant fluid of the Submerged mycelial liquid tissue culture, and adding the collected Supernatant (56) References Cited fluid to a food product in an amount Sufficient to enhance the food products taste. In one embodiment, the mycelial liquid U.S. PATENT DOCUMENTS tissue culture comprises C. Sinensis, and the culture step is carried out for between about one and sixty days. The food 1,822.227 A 9, 1931 Lendrich et al. products include quinoa, amaranth, whole wheat, Stevia 2.693,664 A 8, 1949 SZuecs plant parts, Steviol glycoside, aspartame, acesulfame-K, 3,749,584 A 7, 1973 Kurtzman et al. Sucralose, carbohydrates, salt Substitutes Such as potassium 3,810,997 A 5, 1974 Chien chloride, monk fruit, cacao, cacao liquor, tea, ginseng, pea 4,590,160 A 5/1986 Nishihashi et al. protein, Sugar alcohol, coffee, cranberry, grapefruit, pome 4,891,220 A 1/1990 Donzis granate, coconut, wine, beer, liquor and spirits. The present E. i A 28 R. invention also includes products made by the disclosed 6,277.396 B1 8/2001 Dente methods. 6,476,003 B1 1 1/2002 Jordan et al. 15 Claims, No Drawings US 9,572,364 B2 Page 2 (56) References Cited JP 2011-103901 A 6, 2011 KR 1487724 B1 2, 2015 U.S. PATENT DOCUMENTS NL 7322 C 4, 1921 WO WO O1/032830 5, 2001 2008/003 1892 A1 2/2008 Kristiansen WO WO 2006,107208 10, 2006 2008/0057162 A1 3f2008 Brucker et al. WO WO 2007/031186 3, 2007 2008/0107783 A1 5/2008 Anijs et al. WO WO 2011/O1268O 2, 2011 2008/0171104 A1 7, 2008 Zhu WO WO 2011/032244 3, 2011 2008/0193595 A1 8/2008 De Vuyst et al. WO WO 2011, 151831 12/2011 2008/0264858 A1 10, 2008 Stamets WO WO 2013, 171194 11 2013 2008/0274234 A1 11, 2008 Miller WO WO 2014f145256 A1 9, 2014 2008/0296,223 A1 12, 2008 Hiromoto WO WO 2014f145265 A2 9, 2014 2008/0299645 A1 12/2008 Holliday WO PCT/US16, 19929 2, 2016 2009/0047236 A1 2/2009 Stamets WO WO 2016/033241 3, 2016 2009.0053363 A1 2, 2009 An WO WO 2016, 138476 9, 2016 2009/0098244 A1 4/2009 Schatzmayr et al. 3988. A 3: Nina OTHER PUBLICATIONS 2009, 0220645 A1 9, 2009 Martinez et al. "Eating Mycelium” (2014) wisegeek.com Conjecture Corporation. 2009/0280212 A1 11/2009 Sugimoto et al. Encyclopedia Britannica, Louis Pasteur, Datasheet online. Copy 2010.0055241 A1 3/2010 Nakano et al. right 2014 Encyclopedia Britannica Inc. retrieved on Feb. 6, 2014). 2010, 0183765 A1 7, 2010 Laan Van Der et al. Retrieved from the Internet: <URL: http://www.britannica.com/ 2010/0203189 A1 8/2010 Holliday Ebchecked/topic/445964/Louis-Pasteurs. Specif. p. 3. 2010,2010/0203194 0221385 A1 9,8/2010 2010 MatsuiSalminen et etal. al. English translation.deOliverir, B. etal. WO 2011/032244A1. Flours 2010/0239711 A1 9, 2010 Li produced from fungus myceliated grain. Published Mar. 24, 2011. 2010/0266726 A1 10/2010 Ogura et al. pp. 1-24. Specif. pp. 2, 6-7. 2010/0316763 A1 12, 2010 Choi et al. Firenzuoli et al. (2008) “The Medicinal Mushroom Agaricus blazei 2011 0008384 A1 1/2011 Stamets Murrill: Review of Literature and Pharmaco-Toxicological Prob 2011/0052758 A1 3f2011 Greiner-Stoeffele lems' Evid. Based Complement Alternat. Med. 5(1): 3-15. 2011 0070332 A1 3/2011 Bernaert et al. Foster (2014) “What is Mycelium?' wisegeek.com. Conjecture 2011/0081448 A1 4/2011 Dunphy et al. Corporation. 392 A. 3. EA's al. Han (2005) Solid-state fermentation of cornmeal with the basidi 2011 O189220 A1 8/2011 Yang omycete Ganoderma lucidum for degrading starch and upgrading 2011 (0200551 A1 8, 2011 Stamets nutritional value J. Appl. Micro. 2005, 99: 910-915. 2011/0206721 A1 8, 2011 Nair Hashim, Puziah (1997) “Effect of Processing on Flavour Precursors, 2011/02296.16 A1 9/2011 Anijs et al. Pyrazines and Flavour Quality of Malaysian Cocoa Beans”. PhD 2011 O262593 A1 10, 2011 Binggeli et al. thesis, Universiti Pertanian Malaysia. 2012fOO278892011/0268980 A1 11/20112/2012 PortellaKalisz et al. Ikrawan,van, Yusepusep (2003) Inflniluence off Carbarboxypeptidases tid on CCocoa 2012.0034339 A1 2/2012 Giuliani et al. Specific Aroma Precursors and Methylpyrazines in Under-Fer 2012.0034344 A1 2/2012 Menon et al. mented Cocoa Beans. PhD thesis, Universiti Putra Malaysia. 2012,0082754 A1 4/2012 Holliday Ishikawa et al. (2001) "Antimicrobial Cuparene-Type 2012/010O249 A1 4/2012 Laan et al. Sesquiterpenes, Enokipodins C and D, from a Mycelial Culture of 2012/O128823 A1 5, 2012 Camu Flammulina velutipes' J. Nat. Prod. 64(7): 932-934. 2012/0171308 A1 7/2012 Da Luz Moreira et al. Kang (2005) Abstract of “Studies on chemical constituents in the 2012/0177781 A1 7/2012 Hayashi -- - - 2012fO190093 A1* 7, 2012 Fukuda .................... C12N 1.14 mycelia from fermented culture of Flammulina velutipes' Zhong 435,165 guo Zhong Yao Za Zhi 30(30): 193-195. 2012/O231114 A1 9, 2012 Bezerra De Oliveira et al. Kang (2003) Abstract of “Studies on chemical constituents of the 2012fO244254 A1 9, 2012 Takahashi mycelia from fermented culture of Flammulina velutipes' Zhong 2012/0321744 A1 12/2012 Chhun et al. guo Zhong Yao Za Zhi 28(11): 1038-1040. 2013/0209608 A1 8, 2013 Berends et al. Konno et al. (2002) “Anticancer and Hypoglycemic Effects of 2013/0209609 A1 8, 2013 Moreno et al. Polysaccharides in edible and Medicinal Maitake Mushroom 2013/03371 14 Al 12/2013 Binggeli et al. Grifola frondosa(Dicks.:Fr.) S. F. Gray” International Journal of 2014/O105928 A1 4/2014 Stamets Medicinal Mushrooms 4(3): 10-21. 2014/0170264 A1 6/2014 Kelly et al. Kuo et al. 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    Process Biochemistry 36 (2000) 1–7 www.elsevier.com/locate/procbio Characterization of lipoxygenase activity from a partially purified enzymic extract from Morchella esculenta Barbara Bisakowski a, Avtar S. Atwal a,*, Selim Kermasha b a Food Research and De6elopment Center, Agriculture and Agri-Food Canada, 3600, Bl6d. Casa6ant West, St-Hyacinth, Que´bec, Canada J2S 8E3 b Department of Food Science and Agricultural Chemistry, McGill Uni6ersity, 21 111 Lakeshore, Ste Anne de Belle6ue, Que´bec, Canada H9X 3V9 Received 22 November 1999; received in revised form 29 February 2000; accepted 3 March 2000 Abstract A crude extract (FI) from Morchella esculenta was partially purified using ammonium sulphate precipitation at 0–60, 60–80 and 80–100% of saturation to obtain fractions (FIIa), (FIIb) and (FIIc), respectively. The highest specific lipoxygenase (LOX) activity was obtained in fraction (FIIa) which showed a recovery of 12.8% and a 2.5-fold increase in purification. Partially purified LOX extract exhibited optimal activity at the acidic pH of 3.0 and showed 62.5% of the maximal activity in the pH range of 4.0–6.0 and less than 12.5% activity in the pH range of 7.0–10.0. Kinetic studies indicated that the LOX activity of fraction (FIIa) m −1 −1 −4 had a Vmax of 0.314 mol hydroperoxide mg protein min and a corresponding Km value of 1.59×10 M. The enzymic activity exhibited a strong specificity towards linoleic acid as substrate while only 29% activity was observed using linolenic acid and approximately 11% was obtained with mono-, di- and trilinolein; however, LOX activity showed a relatively strong affinity (83%) towards arachidonic acid as substrate.
  • Plant List: M.E. Depalma Park Picture Botanical Name Common Name Picture Botanical Name Common Name Salvia Coccinea Tropical

    Plant List: M.E. Depalma Park Picture Botanical Name Common Name Picture Botanical Name Common Name Salvia Coccinea Tropical

    Plant List: M.E. DePalma Park Picture Botanical Name Picture Botanical Name Common Name Common Name Salvia coccinea Muhlenbergia capillaris Tropical Sage (red) pink Florida Muhly Grass Blooms: Year-round Blooms: Mid Summer - Fall Attracts Butterflies, Hummingbirds and Painted Buntings Florida Native Florida Native Psychotria nervosa Arachis glabrata Wild Coffee Perennial peanut Blooms: Spring-Summer Fruits: Fall Blooms: Year-round Berries are a food source for birds Native to Argentina, Brazil and Paraguay Male Butterflies need the alkaloids found in this plant, in order to mature sexually. Florida Native Callicarpa americana Gaillardia pulchella Beautyberry Blanket Flower Blooms: late Summer Fruits: Fall Blooms: Year-round Berries are a food source for birds Butterfly nectar plant Florida Native Florida Native Chrysobalanus icaco Stachytarpheta jamaicensis Cocoplum Blue Porterweed Blooms: Spring-Summer Fruits: Fall Blooms: Year-round Berries are a food source for birds Butterfly nectar plant Florida Native Florida Native Asclepias curassavica Pentas lanceolata Mexican Butterfly Weed, Blood-flower or Scarlet Milkweed Egyptian starflower Blooms: Year-round Blooms: Year-round Butterfly nectar plant for Butterflies especially Queens and These may be red, white, lavender, purple, or shades Monarchs of pink. Some are two-toned. All are extremely Monarch larval plant attractive to butterflies, and the red and dark pink varieties delight hummingbirds. Butterfly nectar plant Mimosa strigillosa Loropetalum Sensitive Plant Chinese Fringe Flower Blooms: Year-round Butterfly nectar plant Butterfly nectar plant Native to Japan and southeastern Asia including Florida Native groundcover southern China. Coreopsis_leavenworthii Coreopsis lanceolata Tickseed Lance leafed coreopsis Blooms: Year-round Butterfly nectar plant Butterfly nectar plant Florida Native Florida Native Lantana involucrata.