Edible foodstuffs and bio reactor design

Abstract

Methods of production of edible filamentous fungal biomat formulations are provided as standalone protein sources and/or protein ingredients in foodstuffs as well as a one-time use or repeated use self-contained biofilm-biomat reactor comprising a container with at least one compartment and placed within the compartment(s), a feedstock, a fungal inoculum, a gas-permeable membrane, and optionally a liquid nutrient medium.

1 . A method for production of filamentous fungal biomass, comprising:
 (a) inoculating a liquid fermentation medium with a filamentous fungus;   (b) culturing the filamentous fungus to form a mycelial biomass;   (c) refreshing at least a portion of the liquid fermentation medium;   (d) culturing the mycelial biomass on the refreshed liquid fermentation medium; and   (e) recovering at least a portion of the biomass.   
     
     
         2 . The method of  claim 1 , wherein step (c) is semi-static. 
     
     
         3 . The method of  claim 1 , wherein step (c) is continuous. 
     
     
         4 . The method of  claim 3 , wherein, in step (c), liquid fermentation medium flows continuously through a vessel containing the biomass. 
     
     
         5 . The method of  claim 1 , wherein the biomass comprises a biomat formed on a surface of the liquid fermentation medium or a membrane associated with the liquid fermentation medium. 
     
     
         6 . The method of  claim 5 , wherein a biomass production rate is at least about 40 grams dry biomass per square meter of the surface of the liquid fermentation medium per day. 
     
     
         7 . The method of  claim 5 , wherein a total biomass yield is about 2.5 kilograms dry biomass per square meter of the surface of the liquid fermentation medium. 
     
     
         8 . The method of  claim 5 , wherein step (e) comprises recovering a portion of the biomat from a mature end thereof. 
     
     
         9 . The method of  claim 8 , wherein step (e) is facilitated by a roller wheel contacting the mature end of the biomat. 
     
     
         10 . The method of  claim 1 , wherein, during or after step (c), a rate of production of the biomass is approximately constant. 
     
     
         11 . The method of  claim 1 , wherein the method is semi-continuous. 
     
     
         12 . The method of  claim 1 , wherein the method is continuous. 
     
     
         13 . The method of  claim 1 , wherein the filamentous fungus is selected from the group consisting of  Fusarium oxysporum  strain MK7 (ATCC Accession Deposit No. PTA-10698),  Agaricus bisporus, Boletus edulis, Cantarellus cibarius, Calvatia gigantea, Cyclocybe aegerita, Ganoderma lucidum, Grifola frondosa, Morchella species, Hypsizygus tessellatus, Hypsizygus ulmarius, Laetiporus species, Lentinula edodes, Pleurotus eryngii, Pleurotus ostreatus, Pholiota microspora, Sparassis crispa , and  Tuber  species. 
     
     
         14 . The method of  claim 1 , wherein the liquid fermentation medium is contained within a tray. 
     
     
         15 . The method of  claim 1 , wherein the liquid fermentation medium is quiescent or static during step (b) and flowing continuously during step (d), or vice versa. 
     
     
         16 . The method of  claim 1 , further comprising inactivating the recovered biomass. 
     
     
         17 . The method of  claim 1 , wherein the step of refreshing is conducted daily. 
     
     
         18 . The method of  claim 1 , wherein the liquid fermentation medium is a liquid synthetic growth medium. 
     
     
         19 . The method of  claim 1 , wherein the liquid fermentation medium comprises a carbon source selected from the group consisting of a sugar, a sugar alcohol, a starch, a starch derivative, a starch hydrolysate, a hydrogenated starch hydrolysate, a lignocellulosic pulp or feedstock, corn steep liquor, acid whey, sweet whey, milk serum, wheat steep liquor, industrial liquor, food refinery products/waste streams, and combinations thereof. 
     
     
         20 . The method of  claim 19 , wherein the carbon source is a lignocellulosic pulp or feedstock and is selected from the group consisting of sugar beet pulp, agricultural pulp, lumber pulp, distiller dry grains, brewery waste, and combinations thereof. 
     
     
         21 . The method of  claim 1 , wherein the liquid fermentation medium comprises a carbon source that is a waste product selected from the group consisting of food waste, by-products of food waste, industrial waste, by-products of industrial waste, agricultural waste, by-products of agricultural waste, plant material, and combinations thereof. 
     
     
         22 . The method of  claim 1 , wherein the liquid fermentation medium comprises a carbon source that is a plant material selected from the group consisting of agricultural crop residues, vegetables, forest products and industry residues, and combinations thereof. 
     
     
         23 . The method of  claim 1 , wherein the liquid fermentation medium comprises a carbon source that is selected from the group consisting of wheat straw, barley straw, rice straw, pea, oat, small grain straw, corn stover, corn fibers, corn fiber gum (CFG), distillers dried grains (DDG), corn gluten meal (CGM), switch grass, hay-alfalfa, sugarcane bagasse, non-agricultural biomass, algal biomass, cyanobacterial biomass, urban tree residue, carrots, broccoli, garlic, potato, beets, cauliflower, softwood first/secondary mill residue, hard softwood first/secondary mill residue, recycled paper pulp sludge, anaerobic digestate, lignocellulosic containing waste, newsprint, waste paper, brewing grains, used rubber tire (URT), municipal organic waste, yard waste, clinical organic waste, sugar, starch, waste oils, olive oils, olive oil processing waste, cricket excrement, waste generated during the production of biofuels, glycerol, and combinations thereof. 
     
     
         24 . The method of  claim 1 , wherein the liquid fermentation medium comprises a carbon to nitrogen ratio of about 7.5 to about 26.9. 
     
     
         25 . The method of  claim 1 , further comprising recovering a product from the recovered biomass. 
     
     
         26 . The method of  claim 25 , wherein the recovered product is selected from the group consisting of organic acids, antibiotics, enzymes, hormones, lipids, mycotoxins, vitamins, pigments and recombinant heterologous proteins. 
     
     
         27 . The method of  claim 25 , wherein the recovered product comprises lipids. 
     
     
         28 . The method of  claim 27 , wherein the lipids are recovered by a process selected from the group consisting of mechanical pressing and solvent extraction.