A method for increasing the production of biomass-degrading enzymes by culturing different fungal strains together under solid-state fermentation conditions followed by liquid-state fermentation.

Abstract

Page 1 of 3 pages. Details of the invention? A name that reflects the invention? A method for increasing biomass digestion enzyme production by culturing different fungal strains together under specific conditions. Solid fermentation followed by liquid fermentation. Five branches of science related to invention. Biotechnology related to methods for increasing the production of biomass digesting enzymes through... Cultivating different strains of fungi together under solid fermentation conditions followed by liquid fermentation. Background in the relevant art or science? Enzymes that digest lignocellulose biomass, namely cellulase and xylanase, are used. It is widely used in various industries worldwide, such as food, animal feed, textiles, and agriculture. The demand for cellulase and xylanase enzymes accounts for 20% of the global enzyme market. Cellulase enzymes consist of three groups: endoglucanase, exoglucanase, and β-. Glucosidase is a group of enzymes that catalyze the breakdown of (1-4)-glycosidic bonds. Within the molecular structure of cellulose, if digestion is complete, glucose is obtained, while the enzyme cytokinesis... 15. Lanase consists of two groups of enzymes: endoxylanase and α-xylosidase, which are enzymes that digest... Hemicellulose is broken down into its smallest units, which are xylose sugars. Biomass digestive enzymes can be produced by culturing fungi through solid fermentation or fermentation. Liquid fermentation versus solid fermentation has the advantage of providing conditions for fungal growth on the biomass surface. The solids, which serve as both nutrients and binding sites, use minimal water and energy; however, fermentation... 20. Liquids also have the advantage of mass transfer in the liquid phase, combined with stirring and mixing, resulting in mass transfer. Fungal cells require adequate nutrients and oxygen for culturing different fungal strains together. One approach to compensating for genome variation between strains of microorganisms is to increase the expression of... Enzymes working together create a synergistic effect in biomass decomposition to produce sugars for growth. And it stimulates the production of enzymes in both strains, including the application of the obtained enzymes in production. 25. Other biological substances next. Although there have been some reports of biomass digestion enzyme production using different fungal strains, there is still no... Cultivating different strains of fungi together under a two-stage fermentation process, in which the fungi are cultured. The potential for different catalytic reactions will result in a diverse group of enzymes that help to enhance... The total output of enzyme production and the products from the action of this group of enzymes, in addition to... 30. The two-stage fermentation process also promotes the synergistic action of fungi under both solid fermentation and liquid fermentation conditions. And liquids, therefore, a method to increase biomass digestion enzyme production by culturing different strains of fungi together? Under conditions of solid fermentation followed by liquid fermentation, a process was developed for batch production. Enzymes that work together efficiently lead to cost reduction, improved quality, and enhanced efficiency. Production of bioproducts from lignocellulose biomass. 35. From searching patent and utility model databases in both Thailand and abroad, the following patents were found: A utility model patent that is similar to this invention is: Korean patent number KR20120082123A describes a mixed enzyme preparation for... Degradation of non-starch polysaccharides by the fungi Aspergillus oryzae GB-N1 and Aspergillus niger GB-X2. Page 2 of 3. By culturing under forced-circulation solid fermentation conditions with controlled moisture content. During fermentation, however, there are limitations as it does not specify the relationship between the two fungal strains to production. Enzyme mixture? Australian Patent No. AU2013299022B2 presents a biomass conditioning method. 5. Lignocellulose is broken down by heating and water under pressure, followed by enzymatic digestion to produce sugar. Xilostat? This patent does not present a method for increasing the production of mixed enzymes. U.S. Patent No. WO2015014364A1 proposes a method for conditioning lignum biomass. Cellulose is broken down using acids and bases combined with heating, followed by enzymatic digestion to produce sugar. Xylose and arabinose? This patent does not present a method for increasing the production of the enzyme mixture. 10 Characteristics and Purposes of Invention This invention involves a process for increasing biomass digestion enzyme production by culturing different fungi. The genetic strains are combined under solid fermentation conditions followed by liquid fermentation in the following steps: The preparation of fungal inoculum with different enzymatic activity compared to the preparation of lignocellulose biomass waste material. From agriculture or industry, solid fermentation is followed by liquid fermentation and processing. 15. Pure enzyme? (partially) This invention aims to increase biomass digestion enzyme production by culturing different fungal strains. The combination of genetic material under solid fermentation followed by liquid fermentation can stimulate... The production of enzyme blends is lower than traditional methods, and the resulting enzyme blends are highly efficient in biodegradation. Cellulose and hemicellulose masses. 20. Complete disclosure of the invention. This invention proposes a method for increasing biomass digestion enzyme production by culturing different fungal strains. The genetic material is combined under solid fermentation conditions followed by liquid fermentation in the following steps: a. Prepare two or more fungal inoculum strains with different enzyme activities by culturing them on... Agar-agar dish consisting of 20% potatoes, 2% dextrose sugar, and 1.5% agar-agar powder. pH. 255.0-5.5 After 7-10 days, transfer the resulting spores to a liquid culture medium containing yeast extract. 0.1% and a surfactant such as Tween-80 at 0.1%. In this step, a fungal inoculum is obtained. Enzyme activity? It's different. B. Prepare lignocellulosic biomass, which is agricultural or industrial waste material, such as: Rice bran, corn husk, palm bunches, coconut husk, etc., are dried in the sun for a period of time. 302-3 days in this stage, resulting in dried lignocellulosic biomass? C. Perform solid fermentation by spraying a liquid fungal culture medium containing spores of both fungi. 911 The biomass strains from step B should produce 10-10 spores per strain. Mix 60-70% moisture content biomass (1.5-2.3 liters per kilogram of biomass) thoroughly and ferment in... A rectangular container with a biomass layer height not exceeding 5 centimeters should be covered with cheesecloth and sprayed with disinfectant. Add 0.15-0.23 liters of water and mix well daily. Ferment at room temperature for 2-3 days. This step is complete. Fungi that grow on biomass. d. Perform liquid fermentation by transferring the solid-state biomass fermented from step b into a mixing tank. Mix and add 40-50 liters of liquid feed per kilogram of biomass, stirring at a speed of 100 revolutions per minute. (Page 3 of 3) Liquid fermentation takes 5-7 days. This stage yields a fermented liquid with high mixed enzyme activity and biomass. Leftover from fungal fermentation. Step J: Filter and separate the fermented biomass from the fermentation liquid. Then, separate the fine sediment from the resulting fermentation liquid. Using a centrifuge for 20-30 minutes, settle the clear liquid with acetone at an acetone to particle ratio. 5 parts clear to 2 parts to 1 by volume. The resulting sediment is partially purified biomass enzyme that can... Can it be reused or benefited from further? What is the best way to invent something? As already mentioned in the section on complete disclosure of the invention;

Claims

OCR09WP07 / 10 / 25691. A method for increasing biomass-digesting enzyme production by culturing different fungal strains together under solid-state fermentation followed by liquid-state fermentation, consisting of: a. Preparing two or more fungal inoculum strains with different enzyme activities and culturing them on agar medium containing 20% ​​w / v potato extract, 2% w / v dextrose sugar, and 1.5% w / v agar powder at a pH of 5.0-5.5 for 7-10 days. The resulting spores were then transferred to the fungal culture medium. a) A culture medium containing 0.1% w / v yeast extract and a surfactant, polysorbate 80 at a concentration of 0.1% w / v, was prepared. This step yielded fungal inoculum with different enzyme activities. b) Lignocellulose biomass, an agricultural or industrial waste material, was prepared. This could be sugarcane bagasse, rice bran, corn cobs, empty palm bunches, or coconut husks. It was sun-dried for 2-3 days, resulting in dry lignocellulose biomass. c) Solid fermentation was performed by spraying the culture medium containing fungal spores onto the biomass from step b.To obtain 10⁹-10⁻¹ spores per kilogram of biomass at 60-70% moisture content (1.5-2.3 liters per kilogram of biomass), thoroughly mix and ferment in a rectangular container with a biomass layer height not exceeding 5 centimeters. Cover with cheesecloth, spray with 0.15-0.23 liters of water, and mix daily. Ferment at room temperature for 2-3 days. At this stage, fungi growing on the biomass are obtained. Next, perform liquid fermentation by transferring the solid-state fermented biomass from step b into a mixing tank and adding 40-50 liters of liquid medium per kilogram of biomass. Stir at 100 rpm and continue liquid fermentation for 5-7 days. At this stage, a fermented liquid with high enzyme activity is obtained, along with the remaining biomass from the fungal fermentation.The fermented biomass was filtered and separated from the fermentation liquid. The resulting liquid was then centrifuged to remove fine sediment for 20-30 minutes. The clear liquid was precipitated with acetone at a ratio of acetone to liquid of 2:1 by volume. The resulting sediment is partially purified biomass-degrading enzyme, which can be further utilized. The fungal strains that produce cellulase enzymes consist of three groups: endoglucanase, exoglucanase, and beta-glucosidase. Xylanase enzymes consist of two groups: endoxylase and beta-xylosidase.