WO2010074610A1 - Procédé de production d'hydrates de carbone par hydrolyse de complexes de polysaccharides issus de plantes aquatiques et variantes - Google Patents

Procédé de production d'hydrates de carbone par hydrolyse de complexes de polysaccharides issus de plantes aquatiques et variantes Download PDF

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Publication number
WO2010074610A1
WO2010074610A1 PCT/RU2009/000714 RU2009000714W WO2010074610A1 WO 2010074610 A1 WO2010074610 A1 WO 2010074610A1 RU 2009000714 W RU2009000714 W RU 2009000714W WO 2010074610 A1 WO2010074610 A1 WO 2010074610A1
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WIPO (PCT)
Prior art keywords
algae
complexes
hydrolysis
carbohydrates
enzymatic hydrolysis
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PCT/RU2009/000714
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English (en)
Russian (ru)
Inventor
Валерий Абрамович ГАЛЫНКИН
Александр Васильевич ГАРАБАДЖИУ
Айрат Хасанович ЕНИКЕЕВ
Original Assignee
Закрытое Акционерное Общество Научно-Исследовательский Институт "Росбио"
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Publication of WO2010074610A1 publication Critical patent/WO2010074610A1/fr

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/14—Preparation of compounds containing saccharide radicals produced by the action of a carbohydrase (EC 3.2.x), e.g. by alpha-amylase, e.g. by cellulase, hemicellulase
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/02—Preparation of oxygen-containing organic compounds containing a hydroxy group
    • C12P7/04—Preparation of oxygen-containing organic compounds containing a hydroxy group acyclic
    • C12P7/06—Ethanol, i.e. non-beverage
    • C12P7/08—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate
    • C12P7/10—Ethanol, i.e. non-beverage produced as by-product or from waste or cellulosic material substrate substrate containing cellulosic material
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
    • Y02E50/00—Technologies for the production of fuel of non-fossil origin
    • Y02E50/10—Biofuels, e.g. bio-diesel

Definitions

  • a method of producing carbohydrates by hydrolysis of polysaccharide complexes of algae (options)
  • the present invention relates to the field of biotechnology, and in particular to a method for producing carbohydrates from algae, which can be used as components of a nutrient medium for the production of ethanol and biologically active substances (BAS).
  • a method for producing carbohydrates from algae which can be used as components of a nutrient medium for the production of ethanol and biologically active substances (BAS).
  • Algae are low-organized plants that lack true roots, stems, and leaves. In our country, the most common brown and red algae. The species composition of our seas is represented by 50% red and 45% brown algae. Plants contain much more carbohydrates in tissues than animals. Seaweed in a dry fat-free state contains from 40 to 82% carbohydrates. Carbohydrates include mono, di and polysaccharides, mucopolysaccharides and complex polysaccharides.
  • Polysaccharides mainly consist of pentoses (arabinose, xylose, ribose and dosoxyribose) and hexoses (glucose, mannose, galactose, fucose.)
  • pentoses arabinose, xylose, ribose and dosoxyribose
  • hexoses glucose, mannose, galactose, fucose.
  • carbon-containing compounds of aquatic plant materials can potentially be converted to monocarbohydrates using a complex of enzymes or complex enzymatic preparations of plant or microbial origin.
  • various monosaccharides or polysaccharides can be used to obtain biologically active substances, food and feed products or as substitutes petrochemical products in the production of organic chemical compounds.
  • the most active chemical catalysts for the hydrolysis of plant cell polysaccharides are mineral acids and alkalis.
  • lignin When processing plant materials with acid, lignin remains insoluble, while cellulose and hemicellulose are hydrolyzed.
  • acid hydrolysis in addition to low molecular weight carbohydrates, lignin and toxic components are formed - furfural, methylfurfural, methanol, formaldehyde and low molecular weight acids and esters, which make the production environmentally dirty and dangerous for environment and, in particular, for staff and residents of nearby regions.
  • lignin is the most difficult problem that has not been resolved so far.
  • 30% (of the initial plant material) of the solid remains in the form of lignin impregnated with sulfuric acid.
  • the lignin is removed from the apparatus directly into the open apparatus - decantation, i.e. lignin, sulfuric acid, methanol, acetic acid, furfural, methylfurfural and other low molecular weight volatile toxic products enter the atmosphere. All of the above led to the closure of enterprises based on chemical hydrolysis of plant materials.
  • US 7,262,331 describes a method for producing liquid fuel from pulp, which is subjected to a high temperature of 280 ° C and then a high pressure of 100-250 bar to obtain a product. In this case, the breakdown of both proteins and polysaccharides occurs, but the breakdown is erratic, therefore they cannot be used as a source of monocarbohydrates.
  • the closest analogue adopted for the prototype is a method of liquefying and saccharification containing polysaccharides of biomass described in application EA200701127.
  • the known invention relates to a method of liquefying and saccharification containing polysaccharides biomass having a relatively high the dry matter content is higher than 20%, and preferably consisting of relatively large fibers and particles, with such a distribution of the size of the fibers and particles, in which at least 20% (mass / mass.) biomass is in the range of 26-70 mm
  • the method is applicable for the liquefaction and saccharification of polysaccharide-containing biomasses, mainly consisting of starch, purified starch, cellulose, hemicellulose and lignin, for example grain or wheat straw.
  • lignocellulosic biomasses are pretreated by exposure to temperatures from PO to 250 ° C for 1-60 minutes, which ensures the availability of cellulose for enzymes and at the same time provides a limited content of fermentation inhibitors in the pretreated biomass.
  • the present invention combines enzymatic hydrolysis based on a combination of hydrolytic enzymes, including an enzyme that causes the hydrolysis of carbohydrates, and an oxidizing enzyme, with stirring based on the principle of gravity, providing mechanical forces, mainly shear and tensile forces, to biomass.
  • Preferred types of mixing are, for example, mixing in free-fall mixers, such as drum-type mixers, tilting mixers or similar mixing devices.
  • this method has been developed with reference to plant objects containing starch as a spare polysaccharide, the content of which can reach up to 50-60%, or biomass containing cellulose, hemicellulose and lignin.
  • biomass is first pretreated with high temperature from 110 ° C to 250 ° C for 5-15 minutes, and then enzymatic treatment with oxidative and proteolytic enzymes, which leads to destruction of the cell wall, in particular, the cell wall proteins.
  • This allows cellulase to access fibrils of cellulose and amylase to starch. In this case, the bonds of lignin with hemicellulose and pectin with cellulose are not broken.
  • This processing method is accompanied by the oxidation of polysaccharide chains to produce organic acids, as well as the formation of aromatic toxic compounds.
  • the processing of biomass at high temperature leads to caramelization of carbohydrates, the destruction of peptides and sulfur-containing amino acids.
  • access to cellulose is provided only by 40-50%, and the rest of the cellulose remains unhydrolyzed and inaccessible to enzymes, but this processing method allows you to completely hydrolyze starch.
  • the disadvantages of the prototype are: large losses of raw materials due to the oxidation of part of the formed glucose into organic acids, when processing a cellulose-lignin complex, cellulose and pectin are not completely released from their connection with hemicellulose and lignin, which leads to incomplete hydrolysis of cellulose and hemicellulose to monosaccharides; ⁇ the formation of lignin saturated with sulfuric acid, which has no use and is released into the environment, causing harm to the environment; on such hydrolysates it is possible to obtain only technical and feed, but not food products. In addition, in order to continue to use hydrolysates as a component of the nutrient medium, it is necessary to introduce the stage of purification from toxic products - aromatic compounds.
  • the objective of the invention is to develop an environmentally friendly and economical method for producing carbohydrates from polysaccharide complexes of algae, which could then be used not only for the production of ethanol and biologically active substances, but also for the production of food products.
  • Analysis of the chemical composition of algae shows that, first of all, they are a source of various biologically active substances (BAS). So, in terms of vitamin content, kelp and fucus algae are 100-1000 times higher than land plants.
  • Most marine plants are represented by a group of algae, which, as a rule, have a relatively simple structure. Algae are microscopic unicellular plants, but individual specimens can reach a length of more than 80 m. Unlike terrestrial plants, in which photosynthesis occurs only in certain parts of plants - in foliage, then the entire surface of algae takes part in photosynthesis in algae.
  • algae Due to the simple structure and large ratio of surface area to volume, algae effectively absorb solar energy, so the productivity of some algae is much greater than the productivity of land plants.
  • the structure of the cell wall has a lot in common with algae and terrestrial plants, but they differ in the polysaccharide content.
  • the water content in algae is 85-90%, algae solids contain 52-65% organic matter, i.e. lkg of dry algae contains 300-400 g of polysaccharides.
  • algae we do not find starch as spare products, but homo- and heteropolysaccharides and sulfated polysaccharides (agar, carrageenan) of various compositions.
  • the cell wall of algae contains only cellulose, hemicellulose and pectin, while in terrestrial plants, lignin is also part of the cell walls, the content of which is up to 30%.
  • Organic matter of brown algae consists of carbohydrates (73-74%), nitrogenous substances (5-15%), lipids (1-3%), pigments and vitamins.
  • carbohydrates 73-74%
  • nitrogenous substances 5-15%)
  • lipids 1-3%)
  • pigments 1-3%)
  • vitamins Several types of polysaccharides were found in various algae: cellulose (35% of the carbohydrate content), hemicellulose (15%), laminaran (10-15% of carbohydrates), alginic acids (15-20%), sulfated glucans - fucoidans, agar and carrageenans (their content can be up to 20-30%) and mannitol.
  • Sulfated polysaccharides after separation of sulfate groups under the action of enzymes - desulfatases, like other polysaccharides, after hydrolysis with hydrolytic enzymes form monosaccharides - glucose, xylose, mannose,
  • kelp contains up to 15-20% of substances that are easily subjected to enzymatic hydrolysis to hexoses and pentoses.
  • algae is a good carbohydrate substrate, which, after preliminary enzymatic hydrolysis of mannitol, cellulose, hemicellulose, pectin, will contain monosugar, which are well absorbed by microorganisms.
  • seaweed is a complete nutrient medium, which contains both a source of carbon and organic nitrogen (amino acids).
  • algae are used as polysaccharide complexes of algae, for example, brown algae (Lamipagiales jaropisa, L. Dacticara, Fusus sp.), red (Ahpfeltia ricata, Fugcellaria tastigiata), green .;
  • reaction of chemical hydrolysis is carried out at a temperature of 50-60 ° C;
  • enzymatic hydrolysis reaction is carried out at a temperature of 45-80 ° C;
  • a complex is used that contains Ultraflo L (NovoZoumes company) - a thermostable multi-active drug containing cellulase, glucanase, xylanase and pectinase, and Allzime D (OLTEX company) and Ceremix (Novozoumezlu) exuza fungal endo B-gluconase. These drugs are introduced in a ratio of 0.1: 0.1: 0, Ir each per 100 g of algae.
  • Example 1 Enzymatic hydrolysis on a laboratory scale.
  • Example 2 Enzymatic hydrolysis on a laboratory scale.
  • the feed was treated with perhydrol (H 2 O 2 ) for 30-40 minutes. at a concentration of 0.3-0.5% H 2 O 2 .
  • the volume of water supplied is equal to the ratio of algae: water - 1: 5.
  • an aqueous solution of the enzyme preparation cellolux, dystism, viscoflo (the ratio of enzymes: algae 0.1: 0.1: 0.1 g per 100 g of algae) is supplied with pH 5.0 +/- 0.3.
  • the process is carried out with stirring for 120 min at 50 ° C +/- 5 ° C and pH 5.0 +/- 0.3.
  • Example 3 Enzymatic hydrolysis on a pilot scale.
  • the hydrolysis process is carried out at a temperature of 50 +/- 5 ° C, pH 5.0 +/- 0.3 for 120 minutes, with a rotating drum. During this period, hemicellulase and xylanase undergo enzymatic hydrolysis of hemicellulose and pectin components to mannose, galactose, arabinogalacturonic and mananuronic acids. Hydrolysis of these polysaccharide groups allows the release of cellulose fibrils.
  • the next step is the action of cellulase hydrolysis of cellulose to disaccharides and monosugars (glucose). At the same time, under the action of desulfokinase, desulfation of the carrageenan polysaccharide, which is subsequently hydrolyzed to galactose.
  • a volume of water of 12 m 3 was poured and the drum was turned on for 15-20 minutes.
  • the drum was stopped and the liquid phase was poured into a receiving tank to the drained liquid phase.
  • the resulting solution of monosaccharides was used as a source of monosaccharides at the fermentation stage during the cultivation of microorganisms.
  • the complete technological process in the pilot plant confirmed the above experimental data in Example 2.
  • the content of reducing substances increased by 3.0 times, i.e. the amount of carbohydrates was 5.0 g / l.
  • the amount of carbohydrates was 5.0 g / l.
  • glucose, manose, galactose and fucose were identified by gas chromatography. All of these carbohydrates are well utilized during growth by microorganisms.
  • the use of the material obtained by this method is possible for the preparation of a nutrient medium for the production of ethanol, antibiotics, enzymes and amino acids, as well as for food and feed additives.
  • the cost of biofuels is 40-50 cents.
  • monosaccharides obtained from algae it will be 15-20 cents per liter.
  • the implementation of this method will reduce the cost of gasoline for vehicles by 1, 5 - 2.0 times.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
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  • General Engineering & Computer Science (AREA)
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Abstract

L'invention se rapporte au domaine des biotechnologies et concerne notamment un procédé de production d'hydrates de carbone à partir de plantes aquatiques, lesquels peuvent être utilisés en qualité de composants de milieux nutritifs afin de produire de l'éthanol et des substances bioactives. L'invention a pour but de développer un procédé respectueux de l'environnement et économique de production d'hydrates de carbone à partir de complexes de polysaccharides de plantes aquatiques qui peuvent par la suite être utilisés non seulement pour la production d'éthanol mais aussi dans la production de produits alimentaires. À cette fin, on propose un procédé de production d'hydrates de carbone par l'hydrolyse de complexes de polysaccharides issus de plantes aquatiques, ceci comprenant une réaction d'hydrolyse en fermentation pour obtenir des hydrates de carbone, et une hydrolyse chimique préalable additionnelle des complexes de polysaccharides de plantes aquatiques à l'aide de perhydrol ( H2O2 ). La réaction d'hydrolyse en fermentation se fait à l'aide de complexes de ferments comprenant la cellulase, l'hémicellulase, la pectinase et la sulfatase.
PCT/RU2009/000714 2008-12-22 2009-12-22 Procédé de production d'hydrates de carbone par hydrolyse de complexes de polysaccharides issus de plantes aquatiques et variantes WO2010074610A1 (fr)

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RU2008151884/13A RU2430114C2 (ru) 2008-12-22 2008-12-22 Способ получения углеводов гидролизом полисахаридных комплексов водорослей (варианты)
RU2008151884 2008-12-22

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RU2460771C1 (ru) * 2011-07-08 2012-09-10 Сергей Семёнович Березин Способ извлечения биологически активных веществ из биомассы одноклеточной водоросли рода chlorella
RU2483644C2 (ru) * 2011-07-29 2013-06-10 Анатолий Анатольевич Хитров Способ получения сухого продукта из бурых морских водорослей и пищевой продукт на его основе (варианты)
RU2731987C2 (ru) * 2018-09-11 2020-09-09 Федеральное государственное автономное образовательное учреждение высшего образования "Уральский федеральный университет имени первого Президента России Б.Н. Ельцина" Способ получения биоэтанола из водорослей

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KR20030067097A (ko) * 2002-02-07 2003-08-14 김형락 스핀고모나스 속 ａｓ6330 균주 및 이를 이용한아가로펙틴 설페타제의 생산방법
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JP2005102639A (ja) * 2003-10-01 2005-04-21 Marutomo Co Ltd 便秘改善用食品およびその製造方法
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