WO2012026713A2 - Procédé d'utilisation de saccharomyces cerevisiae pour produire de l'éthanol à partir de fructane, et composition permettant de produire de l'éthanol - Google Patents

Procédé d'utilisation de saccharomyces cerevisiae pour produire de l'éthanol à partir de fructane, et composition permettant de produire de l'éthanol Download PDF

Info

Publication number
WO2012026713A2
WO2012026713A2 PCT/KR2011/006122 KR2011006122W WO2012026713A2 WO 2012026713 A2 WO2012026713 A2 WO 2012026713A2 KR 2011006122 W KR2011006122 W KR 2011006122W WO 2012026713 A2 WO2012026713 A2 WO 2012026713A2
Authority
WO
WIPO (PCT)
Prior art keywords
ethanol
strain
fructan
saccharomyces cerevisiae
fermentation
Prior art date
Application number
PCT/KR2011/006122
Other languages
English (en)
Korean (ko)
Other versions
WO2012026713A3 (fr
Inventor
최의성
이홍원
전재흥
유병태
정동호
Original Assignee
한국생명공학연구원
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 한국생명공학연구원 filed Critical 한국생명공학연구원
Publication of WO2012026713A2 publication Critical patent/WO2012026713A2/fr
Publication of WO2012026713A3 publication Critical patent/WO2012026713A3/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • C12N1/185—Saccharomyces isolates
    • 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
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/85—Saccharomyces
    • C12R2001/865—Saccharomyces cerevisiae
    • 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

  • the present invention relates to a method for producing ethanol using fructan as a carbon source using Saccharomyces cerevisiae, and a composition for producing ethanol, specifically, a large amount contained in non-grain energy crops.
  • the present invention relates to a composition for efficiently producing bioethanol by utilizing lumptan as a carbon source, a medium containing fructan, and a method for producing ethanol and ethanol-containing food using the same.
  • Biomass resources are the backbone of sustainable bioeconomics, such as biofuels such as bioethanol and biobutanol, and biorefineries for industrial biotechnology.
  • biofuels such as bioethanol and biobutanol
  • biorefineries for industrial biotechnology.
  • starch derived from grains such as corn and sugarcane-derived sugar
  • carbon dioxide reduction effect is known to be insignificant
  • the use of lignocellulosics biomass Development of technology related to the generation of bioethanol is being actively conducted.
  • fructan the second largest stock of energy storage carbohydrates in plants, is promising (Dias-Tagliacozzo et al ., Revta. Brasil. Bot., 22: 267). 273, 1999).
  • Fructan is a polymer in which fructose is polymerized with ⁇ -2,1-bonds or ⁇ -2,6-bonds (DP 2 to 35) and glucose is present at the reducing end.
  • Inulin or levan is known, and inulin is known to be contained in a large amount (about 20%) in tubers such as porridge, Jerusalem artichoke , chicory, dahlia, and yacon.
  • the rubber dandelion ( Russian dandelion) accumulates rubber in the root part, and contains about 25-40% of fructan as a building carbohydrate, which contains fructan when extracting rubber from the root. Large amounts of by-products are produced (van Beilen JB et al., Trends in Biotechnology 25, 522-529 (2007)). Therefore, there is a need for the production of useful substances such as ethanol using fructan produced by such by-products.
  • Pretreatment methods for hydrolyzing fructan to monosaccharides or disaccharides include enzyme treatment (Razmovski R. et al ., Food Biotechnol., 4: 284, 1990) or acid treatment (Negro MJ et al ., Appl. Biochem. Biotechnol. , 129 ⁇ 132: 922 ⁇ 932, 2006), etc., and have been developed by Zymomonas mobilim and Saccharomyces cerevisiae. Method for Enhancing Ethanol Productivity by Mixed Culture with Kluyveromyces fragilis Strains (Szambelan K.
  • ethanol is produced by co-glycosylation process from inulin extracted from swine potatoes using Saccharomyces cerevisiae transformed to express inulinase derived from Kluyveromyces marxianus strain.
  • Saccharomyces cerevisiae transformed to express inulinase derived from Kluyveromyces marxianus strain.
  • beta-fructosidase ⁇
  • Kluyveromyces martianus strain is a Crabtree-negative yeast metabolism biased to the TCA cycle and energy generation, and ethanol from the substrate compared to the Saccharomyces cerevisiae strain. It is known that the ability to make cell mass is more advanced than to make cell mass (Lane MM & Morrissey JP, Fungal Biol. Rev. 24, 17-26 (2010)).
  • the Kluyveromyces marcianus strain is known to have a very low resistance to ethanol compared to the Saccharomyces cerevisiae strain, and despite many studies to date, inulin-containing substrates such as pork potatoes are used. It is known to be the biggest factor that cannot be used in industrial ethanol fermentation process (Chi Z.-M. et al., Bioresour. Technol. 102, 4295-4303 (2011), Lane MM & Morrissey JP, Fungal Biol. Rev. 24, 17-26 (2010)).
  • the present inventors have diligently tried to find a way to produce ethanol by decomposing fructan without pretreatment, and as a result, it has been proved to be harmless to the human body as a generically regarded as safe (GRAS) strain, and has been used for the production of liquor for a long time.
  • GRAS generically regarded as safe
  • the strains of Saccharomyces cerevisiae the strains of Saccharomyces cerevisiae, KCCM 50549, have the ability to resolve the existing Saccharomyces cerevisiae strain or fructo-oligosaccharide.
  • Confirmation of ethanol production ability from fructan with significantly higher efficiency than known Saccharomyces cerevisiae NCYC 625 strain Schor-Galindo S. et al ., Curr. Microbiol., 41: 89-95, 2000
  • the present invention has been completed.
  • One object of the present invention relates to a composition for producing ethanol comprising KCCM 50549 strain in fructan and Saccharomyces cerevisiae.
  • Another object of the present invention relates to a medium composition for producing ethanol comprising fructan.
  • Another object of the present invention is to culture the KCCM 50549 strain in Saccharomyces cerevisiae, in a medium containing fructan as a carbon source; And it relates to a method for producing ethanol comprising the step of recovering ethanol from the medium.
  • Another object of the present invention relates to a method for producing an ethanol-containing food comprising culturing Saccharomyces cerevisiae KCCM 50549 strain with a plant or extract thereof containing fructan.
  • ethanol is efficiently used by using the strain of Saccharomyces cerevisiae KCCM 50549, which can use the fructan contained in fructan, especially non-grain energy crop, pork potato
  • Saccharomyces cerevisiae KCCM 50549 which can use the fructan contained in fructan, especially non-grain energy crop, pork potato
  • it is industrially very economical in the production of bioethanol, which has the potential for application to the biofuel industry.
  • it has the potential for application to ethanol-containing functional foods, there is an advantage that can be used as feed additives, such as the cells contained in the residue after fermentation.
  • Figure 1 shows a graph for the HPLC chromatogram analysis of the composition of Saccharomyces cerevisiae pig potato fermentation supernatant fructo-oligosaccharide (GF2, GF3, GF4 are 1-Kestose, respectively) (1-kestose), nystose and 1-fructofuranosylnystose, with numbers indicating the degree of polymerization (DP) values of fructo-oligosaccharides) .
  • Figure 2 shows the ethanol fermentation results of pork potato powder using Saccharomyces cerevisiae KCCM 50549 strain.
  • Figure 3 shows a graph for the HPLC chromatogram analysis of fructo-oligosaccharide composition of the industrial dandelion fermentation supernatant of Saccharomyces cerevisiae strain (GF2, GF3, GF4 is 1-Kestose (1-kestose, respectively) ), Nystose and 1-fructofuranosylnystose, and numbers represent the degree of polymerization (DP) values of fructo-oligosaccharides.
  • RD 625 is NCYC 625 strain
  • RD 3233 is NCYC 3233 strain
  • RD 50549 is a group treated with KCCM 50549 strain.
  • the present invention provides a composition for producing ethanol comprising fructan and Saccharomyces cerevisiae KCCM 50549 strain.
  • the term 'fructan' in the present invention means a known natural polysaccharide including the carbohydrates levan and inulin.
  • Levan used in the present invention may generally mean D-fructan composed of chains of polyfructose, in which most of the fructose units are linked to each other only by ⁇ (2-6) fructosyl-fructose bonds.
  • the levan may be naturally occurring in a specific plant species (in this case, they are called plains), and those produced through metabolism of specific bacteria may be used, and extracted from a specific plant according to a conventional technique. However, those produced by fermentation techniques and enzymatic in vitro synthesis may be used, but are not limited thereto. Preferably it may be extracted from the root or tuber of the plant raw material such as pork potato, chicory, dahlia, yacon or industrial dandelion.
  • the term 'inulin' generally refers to D-fructan composed of chains of polyfructose in which most of the fructose units are linked to each other by ⁇ (2-1) bonds, and used interchangeably with fructans.
  • Inulin in the present invention may be produced using a bacterium or a plant, or may be formed by enzymatic in vitro synthesis or the like starting from sucrose, but is not limited thereto.
  • the term 'extraction' in the present invention means obtaining without acid treatment and enzyme treatment, and can be extracted according to various extraction methods such as cold needle extraction, heat extraction, ultrasonic extraction, cold extraction as known in the art. .
  • the extraction method is not limited thereto, and may be extracted at room temperature or warmed under conditions in which fructan as an active ingredient is not destroyed or destruction is minimized.
  • it may be dried according to various drying methods such as hot air drying, freeze drying, and vacuum drying.
  • fructan obtained by using a known general extraction method without using acid treatment or enzymatic treatment of swine potatoes, chicory, dahlia, yacon or industrial dandelion can be used. It is also possible to use fructan-containing plants themselves or dried products thereof.
  • the term 'degree of polymerization' (DP) in the present invention means the number of (average) of the basic unit per molecule when the molecules are composed of regular repeating units.
  • the degree of polymerization of fructan used in specific embodiments of the present invention is 1 to 15, preferably 8 to 15, but is not limited thereto.
  • Saccharomyces cerevisiae KCCM 50549 strain' in the present invention is a known natural yeast strain deposited with the Korea Microorganism Conservation Center, and the wild type is diploid, has high ethanol resistance, glucose and The biomass hydrolyzate containing glucose and xylose is characterized in that the strain is capable of producing ethanol as a carbon source.
  • Saccharomyces cerevisiae KCCM 50549 strain is the same strain as Saccharomyces cerevisiae ATCC 24858 strains deposited in the American Type Culture Collection (ATCC), and those skilled in the art It can be obtained and used freely. It was first identified by the inventors that the strain could produce ethanol from fructans.
  • Fermentation time may vary depending on the inoculation amount of Saccharomyces cerevisiae KCCM 50549 strain used in specific examples of the present invention.
  • the present invention provides a medium composition for the production of ethanol comprising fructan.
  • the term 'ethanol production medium composition' in the present invention means a medium composition capable of producing ethanol as a metabolite while growing KCCM 50549 in the strain Saccharomyces cerevisiae including fructan as a carbon source.
  • the medium composition may include a known medium, the culturing process of the strain may be made according to the appropriate medium and culture conditions known in the art.
  • the type of medium used for the cultivation should suitably meet the requirements of the particular strain.
  • the medium composition may include various carbon sources, nitrogen sources, and trace element components in addition to fructans.
  • carbon sources that may be used include carbohydrates such as glucose, fructose, sucrose, lactose, maltose, starch and cellulose, and soybean oil (such as cellulose).
  • fats such as soybean oil, regular sunflower oil, castor oil, coconut oil, palmitic acid, stearic acid and linoleic acid Fatty acids, alcohols such as glycerol and ethanol, organic acids such as acetic acid, and the like. These carbon sources may be used alone or in combination.
  • the fructan contained in the medium composition of the present invention may be extracted from the root or tuber of pork, chicory, dahlia, yacon or industrial dandelion, but is not limited thereto.
  • the medium composition of the present invention may include the fructan-containing plant itself or dried material thereof.
  • the fructan of the present invention is characterized by no pretreatment of enzyme or acid treatment.
  • the medium composition according to a specific embodiment of the present invention may include 1 to 500 g / L of fructan as a carbon source, preferably 100 to 500 g / L, but is not limited thereto. .
  • nitrogen sources examples include organic nitrogen sources and urea (CO (NH 2 ) 2 ), sulfuric acid such as peptone, yeast extract, gravy, malt extract, corn steep liquor (CSL) and soybean wheat.
  • the medium composition may include potassium dihydrogen acid (KH 2 PO 4 ), dipotassium hydrogen phosphate (K 2 HPO 4 ), and a corresponding sodium (Na) -containing salt as a phosphorus source. It may also include metal salts such as magnesium sulfate (MgSO 4 ) or iron sulfate. In addition, amino acids, vitamins, and appropriate precursors may be included.
  • compounds such as ammonium hydroxide, potassium hydroxide, ammonia, phosphoric acid and sulfuric acid can be added to the culture in an appropriate manner to adjust the pH of the culture.
  • antifoaming agents such as fatty acid polyglycol esters can be used to suppress bubble generation.
  • oxygen or oxygen-containing gas eg, air
  • oxygen or oxygen-containing gas injected into the culture to maintain the aerobic condition of the culture.
  • This culture process can be used by those skilled in the art can be easily adjusted according to the type of strain selected.
  • Examples of the culture method include, but are not limited to, batch culture, continuous culture or fed-batch culture.
  • the present invention comprises the steps of culturing Saccharomyces cerevisiae KCCM 50549 strain in a medium containing fructan as a carbon source; And it provides a method for producing ethanol comprising the step of recovering ethanol from the medium.
  • the fructan may be extracted from the root or tuber of pork, chicory, dahlia, yacon or industrial dandelion, but is not limited thereto.
  • the medium of the present invention may include the fructan-containing plant itself or a dried product thereof.
  • the fructan of the present invention is characterized by no pretreatment of enzyme or acid treatment.
  • the term 'culture' in the present invention means to grow under the environmental conditions appropriately artificially controlled microorganisms.
  • the method of culturing the KCCM 50549 strain in Saccharomyces cerevisiae in a medium containing fructan to produce ethanol may be performed using a method well known in the art.
  • the culture may be continuously cultured in a batch process or in a fed batch or repeated fed batch process, but is not limited thereto.
  • Saccharomyces cerevisiae KCCM 50549 strain in the present invention has a feature that can be used as a substrate without making a hydrolyzate after pretreatment such as acid treatment or enzyme treatment of fructan used as a carbon source.
  • the method of culturing Saccharomyces cerevisiae KCCM 50549 strain in a medium containing fructan as a carbon source can be performed according to well-known methods, such as culture temperature, incubation time and pH of the medium. The conditions of can be adjusted appropriately to maximize the production of ethanol.
  • the method for producing ethanol of the present invention includes recovering ethanol produced from a strain or culture medium.
  • the method for recovering ethanol produced from a strain or culture medium uses a method well known in the art.
  • the term 'recovery' in the present invention means to obtain the desired product using any method capable of obtaining the desired product from the strain or culture medium, preferably a mixture of two or more liquids having different boiling points
  • any method capable of obtaining the desired product from the strain or culture medium preferably a mixture of two or more liquids having different boiling points
  • heating the solution to a specific temperature to collect the vaporized gas from the solution to liquefy again to separate the liquid mixture to obtain the desired product
  • Vacuum distillation, steam distillation can be used to obtain the desired product, but is not limited thereto.
  • Saccharomyces cerevisiae KCCM 50549 strain is about 2.1 times as compared to Saccharomyces cerevisiae ATCC 96581 strain, a common ethanol fermentation strain used as a control, and a short chain fructo- Saccharomyces cerevisiae, which is known to use some oligosaccharides (fructo-oligosaccharide), showed about 1.6 times higher ethanol productivity than the NCYC 625 strain.
  • Saccharomyces cerevisiae NCYC 625 strain which is known to utilize some fructo-oligosaccharides, uses only fructo-oligosaccharides with DP values of about 6 to 7 and higher polymerization degrees. While the oligosaccharides having the above were not used, the Saccharomyces cerevisiae KCCM 50549 strain of the present invention was shown to consume most of the fructo-oligosaccharides up to a DP value of 15 (Fig. 1). It was also confirmed that ethanol can be produced with high efficiency from fructans present in industrial dandelion rubber extraction by-products (Table 1).
  • the present invention provides a method for producing an ethanol-containing food comprising culturing Saccharomyces cerevisiae KCCM 50549 strain with a plant containing fructan or an extract thereof. .
  • the method may further comprise the step of removing the Saccharomyces cerevisiae KCCM 50549 strain. Removal of the strain may be performed using a method known in the art, and may be performed after ethanol is completed by culturing the plant or extract thereof with Saccharomyces cerevisiae KCCM 50549 strain.
  • Saccharomyces cerevisiae Saccharomyces cerevisiae ( Saccharomyces cerevisiae ) KCCM 50549 strain of the present invention is a GRAS (generally regarded as safe) strain as a representative alcohol-producing yeast strain proved to be safe for the human body, so has an advantage in the manufacture of food It can be used in various ways.
  • GRAS generally regarded as safe
  • the fructan-containing plant of the present invention may be, but not limited to, pork potato, chicory, dahlia, yacon or industrial dandelion. It may also comprise the fructan-containing plant itself or its dry matter.
  • the fructax-containing plant or extract thereof may be extracted by various methods known in the art, and may be extracted from the roots or tubers of pork, chicory, dahlia, yacon or industrial dandelion.
  • the fructan of the present invention is characterized by no pretreatment of enzyme or acid treatment.
  • the food of the present invention may be used as a dietary supplement, in this case, the medium composition from which the strain is removed may be added as it is, or may be used together with other dietary supplements or dietary supplements, and may be appropriately used according to a conventional method. Can be.
  • the mixing amount of the active ingredient may be appropriately determined depending on the intended use.
  • the type of ethanol-containing food of the present invention is not particularly limited, but examples thereof include beverages, tea, drink, makgeolli, shochu, whiskey or beer, and may include all of the health functional foods in a conventional sense. It may include food used as a feed for.
  • the food of the present invention includes various nutrients, vitamins, electrolytes, flavors, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH regulators, stabilizers, preservatives, glycerin, alcohols. And carbonation agents used in carbonated beverages. Others may contain pulp for the production of natural fruit juices, fruit juice drinks and vegetable drinks.
  • the fructan-containing pork potato powder was purchased from commercially available products, and the content of inulin-type sugars was increased by the glucose and fructose content (4.1% and 51.8%, respectively) obtained by acid hydrolysis. When analyzed, it was analyzed as 55.9%. Pork potato powder was suspended in distilled water to 135 g / L or 180 g / L, adjusted to pH 5.0 and sterilized for 15 minutes, and then used in flask culture and fermenter culture without addition of other nutrient medium.
  • Example 2 Yeast strain used
  • Saccharomyces cerevisiae ATCC 96581, NCYC 625, and NCYC 3233 strains were purchased from ATCC (American Type Culture Collection, Virginia, USA) and NCYC (National Collection of Yeast Cultures, Norwich, UK), respectively.
  • the KCCM 50549 strain in cerevisiae was purchased from the Korean Culture Center of Microorganisms (KCCM).
  • the flask was cultured using a 1 L flask, inoculated with 200 ml of pork potato powder suspension (135 g / L) in 10 ml YPD (Yeast Peptone Dextrose) medium overnight, and then incubated at 30 rpm at 48 ° C. Time shake culture. Fermenter culture was used 5 L fermentation tank (COBIOTECH, Incheon, Korea), and 2 L pig potato suspension (180 g / L) was used to adjust the initial pH to 5.0, followed by incubation in 200 ml of YPD medium. Was inoculated and fermentation was carried out at 30 ° C. under conditions of agitation speed 400 rpm and aeration rate 0.2 vvm.
  • fructo-oligosaccharides sugar composition of the fermentation supernatant was analyzed by HPLC (Shimadzu LC10Avp) using a Shodex Asahipak NH2P-50 column. Distilled water and acetonitrile 35:65 mixed solution were flowed into the mobile phase at 1 ml / min. Column temperature was set to 40 ° C. and the eluted sugar was analyzed using RI detector.
  • Example 5 Saccharomyces cerevisiae strain selection excellent ethanol fermentation ability
  • Saccharomyces cerevisiae strains which efficiently produce ethanol, were searched using a pig potato powder suspension that did not undergo pretreatment such as acid treatment or enzyme treatment.
  • pretreatment such as acid treatment or enzyme treatment.
  • the short chain fructo-oligosaccharides contained in Saccharomyces cerevisiae ATCC 96581 strain or pork potato, which is known as the conventional ethanol fermentation strain were found in Saccharomyces cerevisiae. oligosaccharides) (Schorr-Galindo S. et al Curr. Microbiol., 41: 89-95, 2000) Saccharomyces cerevisiae was found to have significantly higher ethanol production capacity than the NCYC 625 strain.
  • Saccharomyces cerevisiae KCCM 50549 strain is about 2.1 times higher than Saccharomyces cerevisiae ATCC 96581 strain, a common ethanol fermentation strain used as a control, and fructo-oligosaccharide of short chain. Saccharomyces cerevisiae, which is known to use some, has a ethanol productivity of about 1.6 times higher than the NCYC 625 strain.
  • Example 7 Ethanol Fermentation of Pork Potato Powder Suspension Using KCCM 50549 Strain in Saccharomyces cerevisiae
  • Saccharomyces cerevisiae was subjected to the ethanol fermentation of a suspension of pork potato powder in a 5 L fermenter using KCCM 50549 strain. As shown in FIG. 2, the fermentation proceeded rapidly and the fermentation was almost completed within 36 hours, and the final ethanol concentration at 48 hours reached 37.3 g / L. This figure corresponds to about 73% of the theory of fermentation conversion yield of inulin-type sugars.
  • KCCM 50549 strain in Saccharomyces cerevisiae does not require pretreatment for hydrolysis of fructan, has the property of utilizing fructan, and proved to be safe for humans as a generically regarded as safe (GRAS) strain. It can be expected to have various industrial applications as a non-GMO strain since it has an advantage as a representative mainstream yeast strain, and also does not require recombinant expression of inulin degrading enzymes.
  • Example 8 Ethanol Fermentation Using Industrial Dandelion Rubber Extract By-Product
  • Saccharomyces cerevisiae KCCM 50549 strains were compared with Saccharomyces cerevisiae NCYC 625 and NCYC 3233 strains for the production of ethanol from industrial dandelion hydrothermal extracts. The results are shown in Table 1 below.
  • the Saccharomyces cerevisiae of the present invention was about 1.7-fold and 2.1-fold higher in ethanol when the KCCM 50549 strain was compared with other Saccharomyces strain NCYC 625 and NCYC 3233 strains, respectively. It can be seen that.
  • the hydrothermal extract obtained from the industrial dandelion root was found to contain fructan up to about 15 degrees of polymerization (DP).
  • DP degrees of polymerization
  • the fructans of DP 6 or higher contained in the hydrothermal extract were not used well, whereas the Saccharomyces cerevisiae KCCM 50549 The strain was used almost all the fermentation up to DP 15, indicating that the trend is consistent with the ethanol production value obtained in the previous fermentation.
  • Example 9 Ethanol Fermentation Using Raw Pork Potato Extract
  • Saccharomyces cerevisiae KCCM 50549 strain was used as a substrate for extracting the raw pork potato heated and compressed to carry out ethanol fermentation using raw pork potato as a raw material. Ethanol fermentation was performed using.
  • the ethanol concentration was 60.7, 61.4 and 60.0 g / l at 24, 48 and 72 hours of the fermentation supernatant, respectively, and the fermentation was completed within 24 hours after the start of fermentation.
  • the fermentation rate was increased and the ethanol content was about 1.7 times higher compared to the ethanol fermentation pattern and content (36.2 g / l after 36 hours) obtained when fermented with dried pork potato powder in Example 7. It could be obtained.
  • the yield was about 84%, which was much higher than the 70% of Example 7 (Table 2).
  • Chicory ( Cichorium intybus L. ) has been cultivated since BC and has used its leaves and roots as a favorite food, such as food, medicinal and coffee substitutes. Chicory contains various bioactive substances such as sugars, organic acids, alkaloids, triterpenes, sesquiterpenes, and coumarin, resulting in lower blood lipids and glucose levels, lower uric acid levels, and soy sauce. It is known to show various effects such as protective effect (Wang Q. et al., Zhongguo Zhong Yao Za Zhi 34, 2269-2272 (2009)). According to a recent report (Juskiewicz J. et al. Arch. Anim. Nutr.
  • Chicory is a representative plant with inulin-type fructans, such as pork potatoes. Therefore, the chicory root extract known to have various functionalities was used as a substrate to investigate whether it can be utilized as an alcohol-containing functional beverage by proceeding with ethanol fermentation.
  • Chicory root dried slices were purchased from the herbal market and washed thoroughly. 900 ml of distilled water was added to 100 g of dried chicory root, pulverized with a mixer, centrifuged at 3000 rpm, and the supernatant was taken. This supernatant was autoclave at 121 ° C. for 15 minutes and used for fermentation without the addition of a separate nutrient medium. The sugar concentration of this extract was found to be 56.9 g / l as measured by acid hydrolysis and the pH was about 4.6, which showed relatively acidic pH. Proceeded. Ethanol fermentation using the flask proceeded for 72 hours under the same conditions as in Example 5.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Mycology (AREA)
  • Genetics & Genomics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Botany (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Virology (AREA)
  • Biomedical Technology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne un procédé permettant d'utiliser Saccharomyces Cerevisiae pour produire de l'éthanol à partir de fructane, et une composition permettant de produire de l'éthanol, et particulièrement : une composition utilisant le fructane, présent en grandes quantités dans les plantes énergétiques non céréalières, comme source de carbone pour produire de façon efficace du bioéthanol ; un milieu de culture contenant du fructane ; de l'éthanol utilisant ce dernier ; et un procédé de production d'aliments contenant de l'éthanol. Lorsque la souche de la présente invention est utilisée, une étape de prétraitement peut être éliminée, ce qui permet de contribuer à la simplification et à la réduction des coûts du traitement.
PCT/KR2011/006122 2010-08-23 2011-08-19 Procédé d'utilisation de saccharomyces cerevisiae pour produire de l'éthanol à partir de fructane, et composition permettant de produire de l'éthanol WO2012026713A2 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
KR20100081662 2010-08-23
KR10-2010-0081662 2010-08-23

Publications (2)

Publication Number Publication Date
WO2012026713A2 true WO2012026713A2 (fr) 2012-03-01
WO2012026713A3 WO2012026713A3 (fr) 2012-05-03

Family

ID=45723903

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2011/006122 WO2012026713A2 (fr) 2010-08-23 2011-08-19 Procédé d'utilisation de saccharomyces cerevisiae pour produire de l'éthanol à partir de fructane, et composition permettant de produire de l'éthanol

Country Status (2)

Country Link
KR (1) KR101349290B1 (fr)
WO (1) WO2012026713A2 (fr)

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009125906A1 (fr) * 2008-04-11 2009-10-15 Jeong Rang Lee Système de recyclage du maïs à titre de source énergétique

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009125906A1 (fr) * 2008-04-11 2009-10-15 Jeong Rang Lee Système de recyclage du maïs à titre de source énergétique

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
BIORESOURCE TECHNOLOGY vol. 101, 2010, pages 4395 - 4402 *
BIORESOURCE TECHNOLOGY vol. 102, 24 August 2010, pages 2109 - 2111 *
J. BIOTECHNOL. vol. 150S, November 2010, page S154 *
J. IND. MICROBIOL. BIOTECHNOL. vol. 35, 2008, pages 303 - 311 *

Also Published As

Publication number Publication date
KR101349290B1 (ko) 2014-01-10
WO2012026713A3 (fr) 2012-05-03
KR20120024439A (ko) 2012-03-14

Similar Documents

Publication Publication Date Title
Mussatto et al. Hydrolysate detoxification with activated charcoal for xylitol production by Candida guilliermondii
US8304219B2 (en) Kluyveromyces strains metabolizing cellulosic and hemicellulosic materials
CN104256086B (zh) 采用糟渣类原料发酵生产富dha饲料添加剂的工艺
Li et al. An environment friendly and efficient process for xylitol bioconversion from enzymatic corncob hydrolysate by adapted Candida tropicalis
Hughes et al. Utilization of inulin-containing waste in industrial fermentations to produce biofuels and bio-based chemicals
JP2011525357A (ja) 酵母バイオマスの製造方法
Liu et al. Erythritol production by Yarrowia lipolytica from okara pretreated with the in-house enzyme pools of fungi
CN101939442A (zh) 产生可发酵糖类和醇的无pH调节系统
CN101624584A (zh) 一种内切型木聚糖酶的制备方法
Wang et al. Bioconversion of cellulose and hemicellulose in reed sawdust to xylo-oligosaccharides and L-lactic acid
Ximenes et al. Enzyme production by industrially relevant fungi cultured on coproduct from corn dry grind ethanol plants
US8227220B2 (en) Process for the preparation of ethanol from starch
CN102511650B (zh) 一种利用菊芋糟渣发酵生产蛋白饲料的方法
Alex et al. Bioethanol production from watermelon rind by fermentation using Saccharomyces cerevisiae and Zymomonas mobilis
KR101435582B1 (ko) 최적의 배지 조성을 이용한 글루콘아세토박터 속 kcg326 균주에 의한 박테리아 셀룰로오스의 제조 방법
CN102770526A (zh) 具有高醇产率的嗜热性意大利热厌氧杆菌marato亚种
CN105062908A (zh) 一种高产木糖醇的热带假丝酵母基因工程菌及其应用
WO2012026713A2 (fr) Procédé d'utilisation de saccharomyces cerevisiae pour produire de l'éthanol à partir de fructane, et composition permettant de produire de l'éthanol
BR102016002700A2 (pt) Processo para produção de xilitol a partir de hidrolisado hemicelulósico de torta de macaúba (acrocomia aculeata) e co-produtos de cervejaria, e uso
CN110964706A (zh) 一种纤维素酶、木聚糖酶和果胶酶的制备方法
Yeunyaw et al. Bioconversion of cassava starch to bio-ethanol in a single step by co-cultures of Amylomyces rouxii and Saccharomyces cerevisiase.
Napitupulu et al. Effect of Saccharomyces cerevisiae ATCC 9763 concentration and fermentation time on bioethanol content from corn stover crude cellulose substrate
CN110734868A (zh) 具有高乳酸生产能力之凝结芽孢杆菌rbe4-4分离株及其用途
Yuwa-Amornpitak et al. Comparative Study of Ethanol Production from Cassava Pulp by a Mixed Culture of Amylomyces Rouxii with Zygosaccharomyces Pseudorouxii and Zymomonas Mobilis
KR101425172B1 (ko) 전분을 함유하는 바이오매스로부터 당수율을 향상시키는 방법

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11820140

Country of ref document: EP

Kind code of ref document: A2

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 11820140

Country of ref document: EP

Kind code of ref document: A2