WO2019167371A1 - Clarified sugar cane juice and polyphenol-containing composition - Google Patents
Clarified sugar cane juice and polyphenol-containing composition Download PDFInfo
- Publication number
- WO2019167371A1 WO2019167371A1 PCT/JP2018/044131 JP2018044131W WO2019167371A1 WO 2019167371 A1 WO2019167371 A1 WO 2019167371A1 JP 2018044131 W JP2018044131 W JP 2018044131W WO 2019167371 A1 WO2019167371 A1 WO 2019167371A1
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- extract
- sugarcane juice
- wheat
- juice
- clarified
- Prior art date
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
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C13—SUGAR INDUSTRY
- C13B—PRODUCTION OF SUCROSE; APPARATUS SPECIALLY ADAPTED THEREFOR
- C13B10/00—Production of sugar juices
- C13B10/02—Expressing juice from sugar cane or similar material, e.g. sorghum saccharatum
Definitions
- the present invention relates to clarified sugar cane juice, a method for producing clarified sugar cane juice, a method for producing a polyphenol composition, and use thereof.
- Clarified sugarcane juice In the sugar making process, the harvested sugarcane is cut finely with a cutter or a shredder and then squeezed with a press to obtain a suspended sugarcane juice. Since this suspended sugarcane juice contains many polymer components other than sugar, these must be precipitated and removed. The precipitation of polymer components has heretofore been performed with lime milk. The precipitate produced by the addition of lime milk is removed, and the resulting supernatant (clarified sugarcane juice) is crystallized to produce raw sugar. However, it is generally not possible to cause precipitation with lime milk alone, so in order to obtain a sufficiently clarified sugarcane juice, in addition to lime milk, a precipitation is generally produced using a flocculant. Has been done.
- the clarified sugarcane juice from which the precipitate has been removed by the addition of lime milk can also be used as a culture medium raw material for microorganisms and the like.
- sugarcane juice is treated with lime milk
- phosphoric acid necessary for culturing microorganisms is precipitated, and thus the content of phosphoric acid in the clarified sugarcane juice is low. Therefore, when using it for the culture medium for microorganisms, it becomes necessary to add phosphoric acid separately.
- calcium derived from lime milk precipitates as impurities when microorganisms and the like are cultured
- clarified sugarcane juice treated with lime milk is used as a culture medium for microorganisms
- phosphorus is also used for the purpose of removing calcium. The acid needs to be added separately. This is the same when using molasses that has undergone lime milk treatment.
- sugarcane juice to which flocculant and phosphoric acid are added is used as a raw material for sugar production and microbial culture medium, the obtained sugar and microbial extract cannot be said to be products using natural materials.
- JP2011-109956 describes a method for producing sugar.
- ethanol is added to a sugar solution obtained by squeezing a plant, a precipitate generated by adding ethanol is removed, and sugar is crystallized from the sugar solution from which the precipitate is removed. It is characterized by making it.
- JP-A-4-248999 describes a method for treating a sugar solution.
- the treatment method described in JP-A-4-248999 in recovering sugar from a diluted sugar solution or concentrated sugar solution in a sugar factory, the diluted sugar solution or concentrated sugar solution is heated and then mixed with a colloidal solution of natural clay mineral. The flocs and / or precipitates produced are removed, and then the sugar is recovered by a conventional method.
- This method uses a colloid solution of natural clay mineral to separate and remove organic and inorganic impurities contained in dilute sugar solution or concentrated sugar solution as floc and / or precipitates (special (Paragraph 0004 of Kaihei 4-248999).
- Paragraph 0006 describes that the natural clay mineral is preferably composed mainly of bentonite, and bentonite is clay mainly composed of montmorillonite. Further, it is described that it is preferable to contain a cationic polymer flocculant such as chitosan together with a colloidal solution of natural clay mineral (claim 4, paragraph 0010, etc. of JP-A-4-248999).
- JP 2009-240326 describes a sugar production method.
- the sugar production method described in JP-A-2009-240326 is a sugar production method in which lime milk (lime milk) is added to raw juice (sugar solution), and carbon dioxide gas is blown to precipitate impurities, thereby purifying the raw juice. It uses lime milk produced by pulverizing shells such as oysters and oyster shells and baked products of weathered corals.
- An object of the invention described in the document is to stably supply a high-quality lime raw material to be lime milk in a sugar production method in which lime milk is added to clean raw juice (JP 2009).
- the raw juice material used in the sugar production method is beet (parasite 0001, 0002, etc. in JP 2009-240326 A).
- Clarified sugar cane that does not require the use of non-natural substances such as lime milk and flocculants, and that does not require the addition of phosphoric acid when used as a culture medium, or can suppress the amount added compared to conventional methods. Development of a method for producing juice efficiently has been demanded.
- Polyphenol-containing composition Polyphenol is a general term for components having a plurality of phenolic hydroxy groups in the molecule. It is contained in most plants and over 5,000 types are known. Polyphenols have long been used in foods, cosmetics and the like as perfumes and pigments. Since the 1990s, antioxidative action, hormone promoting action, etc. have been known, and application as health foods and pharmaceuticals has attracted attention.
- Sugar cane juice has also been reported to contain polyphenols.
- US Patent Application Publication No. 2003/0147978 describes a prophylactic and therapeutic agent for infections containing sugarcane-derived extracts as active ingredients.
- This document describes a non-sugar component substance having a molecular weight other than 10,000 derived from a sugarcane extract and exhibiting absorption characteristics for light having a wavelength of 420 nm.
- Japanese Patent Application Laid-Open No. 2003-137803 discloses a method for producing brewed liquor containing sugarcane polyphenols, wherein sugarcane ears are mixed with a raw material of the brewed liquor, and at least sugarcane ears and water.
- the therapeutic formulations of the literature comprise an effective amount of one or more compounds having at least one hydroxyl group and the ability to alter body mass composition, or a physiologically acceptable analog, derivative or prodrug thereof; Comprising a therapeutic formulation comprising an acceptable carrier and altering the body mass distribution by reducing the overall proportion of fat and / or increasing the proportion of non-fat mass to fat mass ( Claim 1).
- a therapeutic formulation comprising an acceptable carrier and altering the body mass distribution by reducing the overall proportion of fat and / or increasing the proportion of non-fat mass to fat mass ( Claim 1).
- sugarcane-derived polyphenols are described as an embodiment of an effective amount of one or more compounds having at least one hydroxyl group and the ability to change body mass composition.
- Example 3 of the document describes the results of examining the polyphenol content of various sugarcane product extracts at different steps of the sugar refining process. The catechin equivalents of squeezed juice, final squeezed juice, syrup, molasses, low-pol sugar, mill mud, shoot apex
- Japanese Patent Application Laid-Open No. 2002-161046 describes a method for producing a sugarcane polyphenol-containing product, wherein sugarcane ears are extracted with water or an organic solvent to extract contained components, and the extract is concentrated to dryness.
- sugar cane ears are extracted with water or an organic solvent, and the components are extracted, the extract is concentrated to dryness, and the aqueous solution of the concentrated dry product is subjected to solvent fractionation or column fractionation.
- a method for producing a sugarcane polyphenol-containing product which is characterized in that it is purified by
- the known methods for obtaining polyphenols from sugarcane juice are limited to direct ethanol extraction, steam distillation, extraction with an organic solvent, adsorption to a hydrophobic resin, and then recovery with ethanol or the like.
- Direct ethanol extraction and steam distillation are inefficient.
- the organic solvent method and the hydrophobic resin adsorption method require special equipment, and cost is required for organic solvent recovery and resin regeneration treatment. Moreover, it cannot be used as a medicine or food if it contains an organic solvent or resin.
- JP2011-109956 JP-A-4-248999 JP2009-240326 US Patent Application Publication 2003/0147978 JP2003-137803 Special table 2008-542307 JP2002-161046
- the present inventors have found that components other than sugar can be efficiently removed from sugarcane juice by using an extract such as beans in a sugar-making process using sugarcane as a raw material, and have come up with the present invention.
- the object of the present invention is to provide a method for producing clarified sugarcane juice.
- the object of the present invention is to provide clarified sugarcane juice.
- the clarified sugarcane juice of the present invention can refine sugar only with natural materials.
- An object of the present invention is to provide a culture medium for culturing microorganisms and the like containing the clarified sugarcane juice of the present invention. Since the culture medium of the present invention contains abundant nutrient sources necessary for culturing microorganisms and the like, the amount of nutrient sources to be attached can be reduced. In addition, since the culture medium does not use an aggregating agent or the like for removing impurities, it can be said that the culture medium is a culture medium made of only natural materials.
- An object of the present invention is to provide a method for culturing microorganisms or cells, comprising culturing microorganisms or cells in a culture medium containing the clarified sugar cane juice of the present invention.
- microorganism or cell may be expressed as “microorganism or the like”.
- An object of the present invention is to provide a microorganism or the like obtained by culturing in a culture medium containing the clarified sugarcane juice of the present invention.
- the present invention aims to provide an extract obtained from the microorganism of the present invention.
- An object of the present invention is to provide a sugar production method.
- the sugar production method of the present invention uses the clarified sugarcane juice of the present invention.
- the present inventors have also found that polyphenols are contained in precipitates from sugarcane juice by using an extract of beans and the like in a sugar making process using sugarcane as a raw material, and have come up with the present invention.
- the present invention aims to provide a method for producing a polyphenol-containing composition.
- An object of the present invention is to provide a polyphenol-containing composition derived from sugarcane juice deposits.
- this invention includes the following aspects [Aspect 1] (1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract, (2) A method for producing a clarified sugarcane juice, comprising obtaining a supernatant of the reaction solution in step (1). [Aspect 2] The production method according to aspect 1, wherein the beans are selected from the group consisting of peas, soybeans, red beans, and peas. [Aspect 3] The method according to embodiment 1 or 2, wherein the wheat is selected from the group consisting of wheat, barley, rye and oats.
- Aspect 8 The method according to any one of aspects 1-7, comprising not adding lime milk, flocculant, or both lime milk and flocculant to sugarcane juice.
- the clarified sugarcane juice according to aspect 9 which does not contain calcium derived from lime milk.
- the phosphate in the material sugarcane juice is substantially maintained.
- a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
- Aspect 13 The culture medium according to aspect 12, for culturing microorganisms.
- Aspect 14 The culture medium for culture according to aspect 12, wherein the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- a method for culturing a microorganism or cell comprising culturing the microorganism or cell in a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
- a method for culturing a microorganism comprising culturing a microorganism in a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
- Aspect 17 The method of embodiment 16, wherein the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, algae.
- a microorganism or cell obtained by culturing in a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
- Aspect 20 The microorganism of embodiment 19, wherein the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- the extract comprises solids at a concentration of 3-15% by weight.
- the present invention efficiently removes non-sugar components contained in sugarcane juice by using an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract. Provide sugarcane juice.
- the clarified sugarcane juice contains abundant nutrients necessary for culturing such as phosphoric acid and vitamins, and thus can be suitably used for culturing microorganisms and the like.
- the fungus bodies such as microorganisms cultured with the clarified sugarcane juice of the present invention and the extract extracted therefrom hardly color, and thus do not affect the color of the food to be added. And since there is no coloring of the wastewater after culture
- the raw sugar and the culture medium produced with the clarified sugarcane juice of the present invention can reduce the amount of phosphoric acid and flocculant added, and the production process can be simplified. Furthermore, by using these raw material sugars and culture medium, it is possible to produce extracts extracted from sugars, microorganisms, etc. using only natural materials as raw materials.
- the method for producing the polyphenol-containing composition of the present invention is an agglomeration method using an extract of beans, wheat and the like, and a normal stirring tank is sufficient except that a device for crushing beans and wheat is required. And polyphenol will accumulate
- FIG. 1 is a comparative photograph after culturing yeast by clarified sugarcane juice culture and molasses culture.
- the left of FIG. 1 is a photograph of molasses culture and the right is a clarified sugarcane juice culture.
- the precipitates in the photograph are cultured cells.
- FIG. 3 is a flowchart of analyzing the polyphenol content by adding a red bean extract to sugarcane juice to obtain a supernatant and a precipitate.
- FIG. 1 is a comparative photograph after culturing yeast by clarified sugarcane juice culture and molasses culture.
- the left of FIG. 1 is a photograph of molasses culture and the right is a clarified sugarcane juice culture.
- the precipitates in the photograph are cultured cells.
- FIG. 2 is a calibration curve for gallic acid.
- FIG. 4 is a flowchart in which a defatted soybean extract is added to sugarcane juice to obtain a supernatant and a precipitate, and a polyphenol content is analyzed (first time).
- FIG. 5 is a flowchart of adding a defatted soybean extract to sugarcane juice to obtain a supernatant and a precipitate and analyzing the polyphenol content (second time). It is the result of having measured DPPH radical scavenging activity using the precipitate obtained from sugarcane juice which added defatted soybean extract.
- the present invention in one aspect, relates to a method for producing clarified sugar cane juice.
- the method for producing clarified sugarcane juice is: (1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract, (2) including obtaining a supernatant of the reaction solution in step (1).
- sugarcane juice is a plant belonging to the genus Saccharum officinarum.
- “Sugar cane juice” is a liquid squeezed from sugar cane. In general, the whole stem or the solid part of the cut sugar cane can be obtained by finely crushing it with, for example, a cutter, a shredder, or a hammer crusher, and then squeezing with a press.
- Extract liquid is liquids extracted from beans and wheat, respectively. Either the bean extract or the wheat extract, or both the bean extract and the wheat extract may be added to the sugar cane juice.
- extract means a bean extract, a wheat extract, or both a bean extract and a wheat extract.
- Beans is a generic term for legume seeds that can be used for food and processing.
- the beans in the present invention are not particularly limited as long as they are seeds of legumes that can be eaten.
- the beans include pea beans, soybeans, red beans, peas beans, kidney beans, chickpeas, empty beans, green beans, and beans.
- the beans are selected from the group consisting of peas, soybeans, red beans and peas.
- the present invention also includes an embodiment using two or more kinds of beans.
- “Wheat” is a general term for gramineous grains.
- the kind of wheat in the present invention is not particularly limited.
- the wheat is selected from the group consisting of wheat, barley, rye and oats.
- the wheat is preferably wheat.
- the present invention also includes an embodiment using two or more kinds of wheat.
- the “wheat extract” is a liquid extracted from the whole wheat plant or a part thereof.
- the “part” of a wheat plant includes leaves, stems, roots, seeds, shoots, preferably seeds.
- this invention also includes the aspect which uses two or more parts of wheat.
- the method for obtaining a “bean extract” or “wheat extract” by extracting a liquid from beans or wheat is not particularly limited. Any method for extracting the liquid can be used depending on the type of beans and wheat used (including the type of used portion of wheat).
- the dried beans or wheat is pulverized with a known pulverizer or the like.
- a solvent such as water or a known buffer solution is added about 5 to 15 times the mass of the dried product.
- the solvent is preferably water or a buffer solution derived from a natural material, and more preferably water.
- the stirring method is not particularly limited and can be appropriately selected by those skilled in the art.
- the conditions such as the type of solvent, pH, temperature, stirring speed, and the like can be appropriately selected appropriately depending on the type of beans or wheat as the material for producing the extract.
- the precipitate is removed from the mixed solution using a known solid-liquid separation method to obtain a supernatant.
- solid-liquid separation methods include gravity sedimentation, centrifugal sedimentation, and filtration. This supernatant can be used as an extract.
- step (1) of the method for producing clarified sugarcane juice the sugarcane juice is added to the bean extract, the wheat extract, or both the bean extract and the wheat extract. Add the extract that is.
- a step of adjusting the pH of sugarcane juice may be included before the addition of the extract in step (1).
- the pH of the sugar cane juice is preferably 2 or more, 3 or more, 4 or more.
- the sugarcane juice pH is preferably adjusted to 9 or less, 8 or less, or 7 or less.
- the sugar cane juice has a pH range of 2-9, 3-9, 4-9, 2-8, 3-8, 4-8, 2-7, 3-7, 4-7.
- the pH of the solvent ranges from 3-9, 3-8.
- acids derived from known natural materials such as citric acid and succinic acid
- bases derived from known natural materials such as sodium carbonate collected in nature, phosphoric acid, sulfuric acid, acetic acid, hydrochloric acid, nitric acid
- pH adjusters such as sodium hydroxide and potassium hydroxide.
- the extract used in step (1) contains solids at a concentration of 1-20 wt%, 3-15 wt%, 4-10 wt%, 4-8 wt%.
- step (1) 1-30% by volume, 1-20% by volume, and 10-20% by volume of the extract are added to the amount of sugar cane juice.
- the concentration of solids derived from the extract in the mixture of sugar cane juice and extract is 0.01-6 wt%, 0.1-4 wt%, 0.1-2 wt%, and 0.0. 5-1% by weight.
- the mixture (reaction solution) is stirred so that it is uniformly mixed.
- the stirring method is not particularly limited and can be appropriately selected by those skilled in the art.
- the mixed solution after stirring the mixed solution, the mixed solution (reaction solution) is allowed to stand.
- the standing time is not particularly limited, but is preferably at least 30 minutes, 1 hour, 1 hour and a half, 2 hours. In one embodiment, the standing time is within 24 hours, within 8 hours, within 6 hours, within 4 hours, within 3 hours.
- the temperature during the standing is not particularly limited.
- the temperature can be appropriately adjusted according to the type of beans or wheat as the material for producing the extract.
- the amount of precipitate during clarification did not change substantially in the range of 40 ° C to 102 ° C. Similar results were obtained for the red bean extract. This means that the efficiency of clarification does not change over a wide concentration range.
- the method for producing the clarified sugarcane juice does not include adding lime milk, a flocculant, or both lime milk and a flocculant to the sugarcane juice.
- “Lime milk” is a white milky suspension obtained by dissolving excess slaked lime (calcium hydroxide) in water with a solubility higher than that of water.
- “Aggregating agent” is a substance added to agglomerate colloidal particles. Examples of typical flocculants include aluminum flocculants such as aluminum sulfate (sulfuric acid band), polyaluminum chloride (PAC), and aluminum chloride, and iron flocculants such as ferric chloride, ferric sulfate, and polysilica iron.
- a polymer flocculant mainly used in combination with an inorganic flocculant is also included in the “flocculating agent” in the present specification.
- the polymer flocculant is cationic, anionic, nonionic and amphoteric.
- the polymer flocculant includes polyamine and the like.
- Calcium milk and flocculant have been considered essential for obtaining clarified sugarcane juice in the sugar making process.
- the method for producing clarified sugarcane juice it is possible to use a bean extract, a wheat extract, or an extract that is both a bean extract and a wheat extract, without using lime milk and a flocculant. It became possible to obtain clear sugarcane juice.
- the sugarcane juice is preferably 40 ° C. or higher, 50 ° C. or higher, 60 You may heat at 70 degreeC or more, 80 degreeC or more, 90 degreeC or more.
- the manufacturing method of clarified sugarcane juice includes the process of obtaining a supernatant liquid from the reaction liquid of a process (1).
- a precipitate is generated.
- the precipitate is removed from the mixed solution using a known solid-liquid separation method to obtain a supernatant. Examples of such a solid-liquid separation method include gravity sedimentation, centrifugal sedimentation, and filtration.
- a sedimentation concentration device for example, a sedimentation concentration device (thickener) can be used.
- a cylindrical centrifuge, a separation plate centrifuge, a decanter centrifuge, or the like can be used.
- a pressure filter such as a leaf filter or a filter press
- a vacuum filter such as a Nutsche type or an Oliver type can be used.
- the heavy liquid of the precipitate first separated by gravity sedimentation or centrifugal sedimentation may be filtered again, and the obtained filtrate may be mixed with the supernatant liquid.
- the supernatant thus obtained is clarified sugarcane juice.
- Clarified sugar cane juice The present invention, in one aspect, relates to clarified sugar cane juice.
- the clarified sugarcane juice is produced by the production method described in “1. Method for producing clarified sugarcane juice”.
- the clarified sugarcane juice preferably does not contain calcium derived from lime milk.
- the clarified sugarcane juice substantially retains the phosphoric acid in the material sugarcane juice. “Substantially maintained” preferably means that the content (% by weight) of phosphoric acid in the clarified sugarcane juice is 80% or more, 85% or more, 90% or more of the content of the material in the sugarcane juice. , 95% or more, 98% or more, or 99% or more.
- the clarified sugarcane juice obtained by adding an extract such as beans preferably has the following characteristics (advantages).
- the use of a flocculant is unnecessary. Therefore, the process for adding lime milk and a flocculant is unnecessary, and it can manufacture efficiently.
- the clarified sugarcane juice obtained by adding an extract such as beans substantially retains the phosphoric acid in the material sugarcane juice. Therefore, since it contains abundant phosphoric acid, the amount of phosphoric acid added can be reduced when used as a culture medium.
- the present invention in one aspect, relates to a culture medium containing the clarified sugar cane juice described in “2. Clarified sugar cane juice”.
- the target to be cultured in the culture medium of the present invention is a microorganism or a cell, but the type is not particularly limited as long as it can be cultured in a culture medium containing sugarcane juice.
- the phrase “can be cultured in a culture medium containing sugarcane juice” means that components contained in sugarcane juice, mainly sugar, are required for growth.
- the present invention relates to a culture medium for culturing microorganisms, comprising the clarified sugarcane juice described in “2. Clarified sugarcane juice”.
- the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- the kind of yeast is not particularly limited as long as it is used for food production.
- yeasts such as liquor yeast, wine yeast, beer yeast and the like can be used.
- the genus Saccharomyces, the genus Shizosaccharomyces, the genus Pichia, the genus Candida, the genus Kluyveromyces, the genus Williopsis It is any one selected from the group consisting of the genus Debaryomyces, the genus Galactomyces, the genus Torulaspora, the genus Rhodotorula, the Yarrowia, and the genus Zygosaccharomyces.
- the yeast is preferably a baker's yeast used for bread production, a torula yeast used for the production of food, feed, etc., or a brewer's yeast used for beer production because of its good growth properties. More preferably, it is a bacterium belonging to Saccharomyces or a bacterium belonging to Candida.
- Saccharomyces include Saccharomyces cerevisiae, Saccharomyces (sake, and Saccharomyces beticus. Saccharomyces sake and Saccharomyces beticus may be classified as a kind of Saccharomyces cerevisiae.
- Examples of the genus Candida include Candida tropicalis, Candida lypolitica, Candida utilis, and Candida sake.
- Saccharomyces cerevisiae any one selected from the group consisting of Saccharomyces cerevisiae, Saccharomyces salmon (Saccharomyces sake), Schizosaccharomyces pombe, and the like.
- Saccharomyces cerevisiae may be used from the viewpoint of abundant eating experience, and Candida utilis may be used from a viewpoint that has a lot of knowledge in research and the like.
- Lactic acid bacteria are a general term for bacteria that acidify lactic acid by metabolism, and are bacteria contained in the order of Lactobacillus. For growth, sugars, amino acids, vitamin B groups, minerals, etc. are required.
- the type of lactic acid bacteria is not particularly limited as long as it is used for food production. For example, bacteria included in the genus Lactobacillus, Enterococcus, Lactococcus, Pediococcus, Leuconostoc, Streptococcus are included.
- Bacillus subtilis is a gram-positive catalase-positive eubacteria that exists universally in soil and plants and exists in ruminants and human gastrointestinal tracts.
- Algae is a general term for organisms that perform oxygen-generating photosynthesis, excluding moss plants, fern plants, and seed plants that mainly live on the ground. In this specification, it means what is a unicellular organism among them.
- the cell is a cell isolated from a multicellular organism, and is selected from the group consisting of animal cells, plant cells, and insect cells.
- the amount of sugarcane juice contained in the culture medium of the present invention is not particularly limited.
- the culture medium comprises sugar cane juice at a concentration such that the sugar concentration is 10-50% weight / volume, 20-40% weight / volume, 28-38% weight / volume. This is the same as the sugar concentration contained in the molasses medium used as a culture medium for microorganisms.
- “Mole molasses” is a by-product generated in the sugar-making process for producing sugar, and is a highly viscous and dark brown liquid. Although it contains 40-60% by weight of sugar, it is difficult to crystallize, so it is removed as a by-product in the sugar production process and used as a raw material for culture medium and livestock feed.
- Molasses as a culture medium for microorganisms is colored by heat treatment in the sugar-making process, and microbial cells cultured using the molasses as a medium and extracts extracted from the microorganisms are also colored. In addition, when such microbial cells and microbial extract are used as a raw material for food, the coloration also affects the color of the food.
- molasses is known to undergo a Maillard reaction over time, and its product causes inhibition of microbial culture and a decrease in fermentation ability. Furthermore, waste water after using molasses for microbial culture is also colored, and the cost of processing the waste water is high, which has been a factor in increasing the selling price of the final product.
- the culture medium containing the clarified sugarcane juice of the present invention can be used for culturing microorganisms or the like, for example, as a medium containing the same sugar concentration and vitamin / mineral amount as the molasses medium.
- the absorbance (OD420), solid content, and chemical oxygen demand (COD) of the culture mother liquor after culture when the culture medium of the present invention was used were examined.
- the absorbance (OD420), solid content, and COD of the culture mother liquor are about 4.8%, about 14.3%, and about 30.5%, respectively, when molasses is used, resulting in wastewater. It became clear that the contamination degree of the culture mother liquor after culturing the microorganisms was extremely low.
- the culture medium does not affect the coloration of the final product when a cell such as a microorganism and an extract such as a microorganism are used as a raw material for food, and the wastewater treatment load can be reduced.
- the “culture medium” can be appropriately adjusted according to the type of microorganism or cell to be cultured except that it contains clarified sugarcane juice.
- the clarified sugarcane juice obtained by adding the extract of beans and the like can be supplied with vitamins and minerals from the extract of beans and wheat. Therefore, in one aspect, preferably, when an extract such as beans is not used, for example, when a molasses medium is used, or when a medium containing sugarcane juice obtained using conventional lime milk is used, The amount of vitamins and minerals added to the culture medium is less than that of the culture medium.
- the present invention relates to a method for culturing microorganisms or cells, comprising culturing in a culture medium containing the clarified sugarcane juice described in “2. Clarified sugarcane juice”. .
- the present invention relates to a method for culturing a microorganism, comprising culturing a microorganism in a culture medium containing the clarified sugarcane juice described in “2. Clarified sugarcane juice”.
- the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- culture medium “microorganism”, “yeast”, “lactic acid bacterium”, “Bacillus subtilis”, “algae”, and “cell” are as described in “3. Culture medium”.
- the culture method of microorganisms or cells can be appropriately selected according to the type.
- Microorganism or cell in one aspect, the present invention relates to a microorganism or cell obtained by culturing in a culture medium containing the clarified sugarcane juice described in “2. Clarified sugarcane juice”.
- the present invention relates to a microorganism obtained by culturing in a culture medium containing the clarified sugarcane juice described in “2. Clarified sugarcane juice”.
- the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- culture medium “microorganism”, “yeast”, “lactic acid bacterium”, “Bacillus subtilis”, “algae”, and “cell” are as described in “3. Culture medium”.
- the culture method of microorganisms or cells can be appropriately selected according to the type.
- the culture medium used for culturing microorganisms or cells of the present invention contains clarified sugarcane juice obtained by adding an extract such as beans to sugarcane juice.
- non-natural additives such as lime milk and flocculants are not used in the manufacture of clarified sugar cane juice. Therefore, it can be said that the microorganism cultured using the culture medium which does not contain such a non-natural product does not contain a non-natural product.
- the present invention relates to a microorganism or cell extract obtained from the microorganism or cell of the present invention.
- An extract is an extract of various components of microorganisms and cells, and includes amino acids, peptides, nucleic acids, minerals, and the like. Moreover, the content ratio of various components can be adjusted according to the types of microorganisms and cells, culture conditions, and extraction conditions.
- the extraction method of the extract is not particularly limited, and any method among methods usually used when extracting an extract from a biological raw material such as a microorganism may be used. Examples of the extraction method include an autolysis method and an enzymatic decomposition method.
- the self-digestion method is a method of solubilizing and extracting a microorganism by the action of an enzyme inherent in the microorganism, and a microorganism extract having a high free amino acid content can be obtained.
- the enzymatic decomposition method is a method in which an enzyme or the like contained in a microorganism is inactivated by heat treatment or the like, and then a decomposition enzyme is added to solubilize and extract the microorganism.
- the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- microorganism “microorganism”, “yeast”, “lactic acid bacterium”, “Bacillus subtilis”, “algae”, and “cell” are as described in “3. Culture medium” and “5. Microorganism or cell”.
- the microorganism of the present invention is a microorganism that is cultured using a culture medium that does not contain a non-natural product and does not contain a non-natural product.
- the microorganism or cell is a microorganism or cell cultured using only natural materials. Such an extract is preferable for use in foods, beverages and the like taken into the living body.
- the present invention relates to a sugar production method as one aspect.
- the sugar production method uses the clarified sugarcane juice of the present system for sugar production.
- Non-limiting sugar production methods include (1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract, (2) Obtaining the supernatant of the reaction solution in step (1), (3) Crystallize sugar from the supernatant obtained in step (2). Including that.
- step (2) the clarified sugarcane juice of the present invention is obtained.
- step (3) a known method for crystallizing sugar from sugarcane juice can be used.
- a method of crystallizing a concentrated solution obtained by vacuum concentration in a honey tank can be used.
- the present invention relates, in one aspect, to a method for producing a polyphenol-containing composition.
- Non-limiting methods for producing a polyphenol-containing composition include: (1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract, (2) including removing the supernatant from the reaction solution of step (1).
- Step (1) is described in “(1) Sugar cane juice”, “(2) Extract” and “(3) Addition of extract to sugar cane juice” in “1. Clarified sugar cane juice production method”. It is as follows.
- the manufacturing method of a polyphenol containing composition includes the process of removing a supernatant liquid from the reaction liquid of a process (1).
- a precipitate is generated.
- the reaction liquid in step (1) is stirred, and then the liquid (supernatant liquid) is removed from the mixed liquid using a known solid-liquid separation method to obtain a precipitate.
- a solid-liquid separation method include gravity sedimentation, centrifugal sedimentation, and filtration.
- “precipitate” and “precipitate” may be used interchangeably.
- a sedimentation concentration device for example, a sedimentation concentration device (thickener) can be used.
- a cylindrical centrifuge, a separation plate centrifuge, a decanter centrifuge, or the like can be used.
- a pressure filter such as a leaf filter or a filter press
- a vacuum filter such as a Nutsche type or an Oliver type can be used.
- the liquid separated by gravity sedimentation or centrifugal sedimentation may be subjected to gravity sedimentation or centrifugal sedimentation again.
- the beans are selected from the group consisting of peas, soybeans, red beans, and peas.
- the wheat is selected from the group consisting of wheat, barley, rye and oats.
- a step of adjusting the pH of sugarcane juice may be included.
- the method includes the step of adjusting the pH of sugarcane juice before adding the extract to a range of 3-9.
- the extract contains a solid content of 3 to 15% by weight.
- step (1) In one embodiment of the method for producing a polyphenol-containing composition, in step (1), 1 to 20% by volume of an extract is added to the amount of sugar cane juice.
- the method includes standing the reaction solution obtained in step (1) for at least 30 minutes before step (2).
- the method for producing a polyphenol-containing composition does not include adding lime milk, a flocculant, or both lime milk and a flocculant to sugarcane juice.
- the polyphenol-containing composition obtained by the production method of the present invention contains polyphenol derived from sugarcane juice.
- the polyphenol-containing composition obtained by the production method of the present invention contains coumaric acid and / or ferulic acid.
- the composition obtained by the production method of the present invention is extracted from the sugar cane juice and beans used in the production method in the state of a wet residue excluding the supernatant liquid in step (2). Containing 15% or more, 20% or more, 25% or more, 30% or more, 35% or more of polyphenol derived from a product.
- polyphenols derived from extracts such as sugar cane juice and legumes migrate to a composition of about 15% to about 40%, about 25% to about 40% sugar cane juice deposits.
- the present invention relates to a polyphenol-containing composition.
- the polyphenol-containing composition is a composition produced by the method for producing a polyphenol-containing composition of the present invention.
- the polyphenol-containing composition contains coumaric acid and / or ferulic acid.
- the polyphenol-containing composition is 15% or more, 20% or more, 25% or more, 30% or more polyphenol derived from an extract such as sugarcane juice or beans used in the production method in a wet state. % Or more and 35% or more.
- the polyphenol-containing composition can be applied to foods and the like as a polyphenol-containing material in a wet state. Polyphenols can be easily extracted with water or ethanol.
- the sugarcane-derived supernatant also contains polyphenol, extraction of polyphenol from the supernatant requires labor and cost of using an adsorption resin or the like.
- the polyphenol-containing composition has antioxidant activity. In one embodiment, it has a high antioxidant activity compared to the same amount of molasses composition.
- the polyphenol-containing composition can be used safely as it is, using sugar cane, that is, only food materials as raw materials.
- a precipitate containing polyphenols can also be obtained by treatment with sugarcane juice lime milk, but the content is relatively low and cannot be used as a food as it is.
- sugarcane itself contains a high concentration of sugar, but the sugar is almost removed by the method for producing a polyphenol-containing composition of the present invention. There is no need for another method for sugar removal.
- the presence of acrylamide has been confirmed in sugar sources that have undergone repeated heating history such as molasses.
- Acrylamide is suspected to be mutagenic and is produced when carbohydrates and the amino acid aspartic acid react when exposed to high temperatures such as baking, frying and roasting (Human exposure and internal doses of acrylamide). food.Food and Chemical Toxicology 43: 365-410).
- high temperatures such as baking, frying and roasting (Human exposure and internal doses of acrylamide).
- the polyphenol-containing composition is further characterized by a low color intensity for the same polyphenol content.
- the polyphenol content with respect to absorbance 1 at a wavelength of 420 nm of a 1% solution may be 0.05 or more, more preferably 0.08 or more, and particularly preferably 0.1 or more.
- Example 1 Preparation of Bean / Wheat Extracts and Sugarcane Juice Dried peas, soybeans (normal soybeans and defatted soybeans), various beans of red beans and peas, various wheats of wheat and barley, and Brown rice and corn are pulverized with a pulverizer such as a mill, and 200 ml of distilled water at 60 ° C. is added to 25 g of the obtained dry pulverized product, followed by stirring for 90 minutes at 500 rpm using a stirrer, followed by centrifugation at 3000 rpm for 5 minutes. went. Thereafter, a supernatant was obtained by decantation. The supernatant thus obtained was used as a legume / wheat extract in the test.
- a pulverizer such as a mill
- Example 2 Addition of various bean and barley extracts to sugarcane juice Various beans and various barleys obtained in Example 1 and brown rice and 45 ml of sugarcane juice prepared to pH 2-9 with potassium hydroxide or phosphoric acid After adding 5 ml of various extracts of corn and stirring gently, the mixture was allowed to stand at 60 ° C. for 1 hour. Thereafter, the supernatant and the precipitate were separated by decantation.
- Example 3 Influence of the addition amount of defatted soybean extract to sugarcane juice
- the defatted soybean extract obtained in Example 1 was boiled and adjusted so that the solid content was 13.5%. Concentrated defatted soybean extract was taken in the amounts shown in Table 2 below, and water was added so that the amount was 2 ml. To this, 13 ml of the sugarcane juice obtained in Example 1 was added, stirred gently, and then allowed to stand at 60 ° C. for 2 hours. Then, it processed at 6000 rpm for 5 minutes with the centrifuge, and isolate
- test no. No. 5 had the highest amount of precipitate.
- Test No. 5 the defatted soybean extract added to sugarcane juice contains 0.12 g of solid content in 2 ml of the extract, and the concentration of solid content in the extract was 6% by weight. And the density
- concentration of the solid content derived from the defatted soybean extract in the defatted soybean extract to be added and the mixed solution was higher than this, the amount of the precipitate was slightly reduced.
- Example 4 Effect of temperature upon addition of various bean extracts to sugarcane juice 5 ml of the defatted soybean extract obtained in Example 1 was added to 45 ml of sugarcane juice prepared to pH 7, and the mixture was lightly stirred and then listed in the table below. Allowed to stand at temperature for 1 hour. Thereafter, the supernatant and the precipitate were separated by decantation.
- the amount of precipitates hardly changed in the range of 40 ° C-102 ° C. Further, 5 ml of the red bean extract obtained in Example 1 was added to 45 ml of sugarcane juice adjusted to pH 5, and after gently stirring, the mixture was allowed to stand at the temperature described in the following table for 1 hour. Thereafter, the supernatant and the precipitate were separated by decantation.
- the amount of the precipitate hardly changed in the range of 30 ° C-90 ° C.
- Example 5 Comparison of yeast culture efficiency of clarified sugarcane juice and molasses treated with beans / wheat extracts
- 180 ml of red bean extract (extracted at 60 ° C., pH 7.5) was added, and room temperature By stirring for 1 hour at (25 ° C.-30 ° C.), the mixture was allowed to stand to obtain a clarified sugar cane juice from which the precipitate was removed as a wet weight from the sugar cane juice.
- Yeast culture was performed using the obtained clarified sugarcane juice.
- the culture apparatus used was BMS-03PI (manufactured by Able), and was cultured in a conventional manner.
- the sugar concentration of sugarcane juice was adjusted to match the sugar concentration (33% w / v) of the molasses medium.
- the yeast used was FT-4 strain (deposit number: FERM BP-808-1) of Saccharomyces cerevisiae.
- the yeast obtained by culturing was treated by a conventional method to obtain a yeast extract, and the nucleic acid content was measured.
- Cell volume The volume of yeast cells obtained by culturing. It measured by putting a 10 ml culture solution into the Spitz glass with a scale, and reading the scale about the volume of the precipitate at the time of centrifuging at 3000 rpm.
- Dry microbial mass (g) Weight when cultivated microbial cells are dried in an oven at 105 ° C. for 4 hours.
- Extract solid content (g) Dry weight of the obtained yeast extract.
- I + G (% by weight) per extract solid content Total weight% of inosinic acid and guanylic acid contained in the extract solid content. It was measured by high performance liquid chromatography (HPLC).
- HPLC high performance liquid chromatography
- Example 6 Comparison of Colors of Clarified Sugarcane Juice Treated with Beans and Wheat Extracts and Yeast Cells Cultured with Molasses Cultured mother liquors of clarified sugarcane juice culture and molasses culture after culturing under the conditions of Example 5
- the absorbance (OD 420 nm) was measured.
- the culture mother liquor was dried using an oven at 105 ° C. for 4 hours, and the weight of the obtained solid content was measured.
- “Solid content (%)” in Table 6 is the ratio (%) of the solid content contained per unit weight of the culture mother liquor.
- the chemical oxygen demand (COD) of the culture mother liquor was measured by a conventional method (JIS K010217).
- FIG. 1 a comparative photograph after yeast culture is shown in FIG.
- the left of FIG. 1 is a photograph of molasses culture and the right is a clarified sugarcane juice culture.
- the precipitates in the photograph are cultured cells.
- FIG. 1 a clear transparent culture solution was obtained by the culture using the clarified sugarcane juice of the present invention.
- Example 7 Analysis of Sugar Cane Juice Precipitate
- sugar bean juice was added with various bean extracts and the components of the resulting precipitate were analyzed.
- the precipitate was further examined for the total amount of polyphenols, p-coumaric acid and total ferulic acid.
- the total amount of polyphenol was measured by a method using a Foreign-Ciocalteu reagent (hereinafter sometimes referred to as “Forin-Thiocarte method”).
- the foreign-thiocult reagent contains phosphotungstic acid as an oxidizing agent, and is measured and quantified by measuring the absorbance at 765 nm by utilizing the fact that it is reduced to a blue color by being reduced by a phenolic hydroxyl group.
- the foreign-thiocult method is well known as a method for measuring the total amount of polyphenols in foods and plants, and is also adopted by ISO (International Organization for Standardization). In this example, MP Biomedicals. A product manufactured by LLC was used, and a calibration curve of gallic acid shown in FIG. 2 was converted as a standard.
- the content of p-coumaric acid was measured as follows. First, 100 ml of methanol was added to 1 g of freeze-dried sugar cane juice precipitate in two portions, and extracted by heating and refluxing at 80 ° C. for 1 hour ⁇ 2 times. The extract was centrifuged and made up to volume with 250 ml. Subsequently, it analyzed by the liquid chromatography tandem type
- the operating conditions of the liquid chromatography tandem mass spectrometer are as follows.
- Total ferulic acid was measured as follows. First, hydrolysis was carried out by adding 3 ml of water and 10 ml of a 1 mol / l potassium hydroxide-ethanol solution to 0.4 g of freeze-dried sugarcane juice precipitate and treating in a boiling water bath for 2 hours. To the reaction, 100 ml of water and 8 ml of 2 mol / hydrochloric acid were added. To this, 50 ml of ethyl acetate was added three times to dissolve in the ethyl acetate layer. After dehydrating and drying, 10 ml of methanol was added. Next, the operating conditions of the high performance liquid chromatography analyzed by high performance liquid chromatography are as follows.
- defatted soybean deposit 1 and defatted soybean precipitate 2 are the results of using sugarcane juice with different harvesting times.
- the deposits obtained from the cane juice using the legume extract contain particularly a large amount of p-coumaric acid.
- polyphenol content in plants other than sugarcane is described in Table 8 (for example, extracted from Table 1 of European Journal of Clinical Nutrition (2010) 64, S112-S120).
- Example 8 Analysis of polyphenols in the supernatant and precipitate of sugarcane juice to which the red bean extract was added Based on the flowchart shown in FIG. 3, the red bean extract was added to the sugarcane juice and the polyphenols in the supernatant and precipitate were The content was examined. The content of polyphenol was measured using the foreign-thiocult method in the same manner as in Example 7 and converted using a calibration curve of gallic acid as a standard.
- BSL 15 A sugar solution in which brown sugar is dissolved in 15% w / v. How to prepare brown sugar Boil sugar cane juice and concentrate while removing the generated starch. The simmering operation is completed with a 5-fold concentration that eliminates water evaporation. Stir well until it turns white so that seed crystals form.
- Example 9 Analysis of polyphenol in the supernatant and precipitate of sugarcane juice to which defatted soybean extract was added Based on the flowchart shown in FIG. 4 or FIG. 5, the defatted soybean extract was added to sugarcane juice, and the supernatant and precipitate were added. The content of polyphenol in the product was examined.
- DFSBE 70 Extract obtained by treating defatted soybeans with distilled water at 70 ° C. for 30 minutes As shown in FIGS. 4 and 5, two tests were performed. In the first round, the total amount of polyphenols in the wet residue was 0.046 g, and the polyphenol recovery rate in the wet residue was 32.81%. The second time, the total amount of polyphenols in the wet residue was 0.021 g, and the polyphenol recovery rate in the wet residue was 15.00%.
- antioxidant activity was measured by DPPH radical scavenging activity using precipitates obtained from sugarcane juice to which defatted soybean extract was added. did.
- DPPH radical scavenging activity In the measurement of DPPH radical scavenging activity, the scavenging activity against an artificial DPPH radical (1,1-diphenyl-2-picrylhydrazyl) is measured with a spectrophotometer. DPPH radicals are purple in the solvent, but when an extract containing an antioxidant is added to this solution, the DPPH radicals are erased and the color becomes lighter. The antioxidant power was evaluated by measuring the absorbance of this color.
- DW distilled water
- DPPH was first sufficiently dissolved with ethanol (0.5 hour-1 hour), and distilled water was added thereto.
- a precipitate obtained from sugarcane juice to which the defatted soybean extract prepared in Example 3 was added was used as a measurement sample. 100 mg of the precipitate was dissolved in 1 ml of 50% ethanol solution, centrifuged, and the supernatant was used for the following measurement.
- a molasses-derived polyphenol-containing composition was used in place of the precipitate.
- the molasses-derived polyphenol composition is prepared by roughly purifying polyphenol from molasses by activated carbon treatment or the like.
- the measurement mixture shown in Table 11 below was prepared, reacted in the dark for 20 minutes, and OD520 nm was measured. Each sample was measured after zeroing with a 50% ethanol solution.
- ID 50 (amount of sample necessary to halve the original DPPH radical) was calculated.
- the precipitate obtained from the sugarcane juice of the present invention has higher DPPH radical scavenging activity than the same weight of molasses-derived polyphenol-containing composition (this example). In the example, it is determined that the ID 50 is about 1/3). That is, the same weight comparison showed that the precipitate obtained from the sugarcane juice of the present invention has stronger radical scavenging activity (antioxidant activity) than the molasses-derived polyphenol-containing composition.
- Example 10 Absorbance of a Solution Containing a Precipitate Obtained from Sugarcane Juice
- an absorbance at 420 nm indicating the yellowness of a solution containing a precipitate obtained from sugarcane juice was examined.
- a precipitate obtained from sugarcane juice a sugarcane juice precipitate (RBE) to which an adzuki bean extract was added and a sugarcane juice precipitate (DFSBE) to which a defatted soybean extract was added were used.
- RBE sugarcane juice precipitate
- DFSBE sugarcane juice precipitate
- MSX-100 Mitsubishi Sugar
- MSX-1MF Mitsubishi Chemicalsui Sugar
- the freeze-dried 4.5 g of RBE and DFSBE were extracted with 15 ml of 50% ethanol aqueous solution.
- the solid content concentration of the extract was 14.76% by mass (% W / W).
- 1% aqueous solution of peanut extract-derived sugarcane polyphenol composition (soluble part) (RBES) and defatted soybean extract-derived sugarcane polyphenol composition (soluble part) ( A 1% aqueous solution of DFSBES) was obtained. Since MSX-100 and MSX-1MF are soluble, a 1% aqueous solution was prepared as it was.
- the total amount of polyphenol was measured by the foreign-thiocult method, and the polyphenol concentration was calculated.
- the polyphenol concentration is shown in terms of gallic acid.
- RBES and DFSBES were shown to have a higher polyphenol content at the same absorbance compared to MSX-100 and MSX-1MF, in other words, a lower color depth of 420 nm at the same polyphenol concentration. .
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
製糖工程では、収穫したサトウキビをカッターやシュレッダーにより細かく切断した後、圧搾機によって搾汁して懸濁したサトウキビジュースを得る。この懸濁サトウキビジュースには、糖以外の多くの高分子成分が含まれているためこれらを沈殿して除く必要がある。高分子成分の沈殿は、従来、石灰乳によって行われてきた。石灰乳の添加によって生じた沈殿物を取り除き、得られた上澄み液(澄明化サトウキビジュース)を結晶化させ、原料糖を製造している。しかしながら、石灰乳のみでは十分に沈殿を生じさせることができない為、十分に澄明化したサトウキビジュースを得るために、石灰乳の他にさらに凝集剤を用いて沈殿を生じさせる、ことが一般的に行われている。 1. Clarified sugarcane juice In the sugar making process, the harvested sugarcane is cut finely with a cutter or a shredder and then squeezed with a press to obtain a suspended sugarcane juice. Since this suspended sugarcane juice contains many polymer components other than sugar, these must be precipitated and removed. The precipitation of polymer components has heretofore been performed with lime milk. The precipitate produced by the addition of lime milk is removed, and the resulting supernatant (clarified sugarcane juice) is crystallized to produce raw sugar. However, it is generally not possible to cause precipitation with lime milk alone, so in order to obtain a sufficiently clarified sugarcane juice, in addition to lime milk, a precipitation is generally produced using a flocculant. Has been done.
ポリフェノールは、分子内に複数のフェノール性ヒドロキシ基を有する成分の総称である。ほとんどの植物に含有され、5,000種類以上が知られている。ポリフェノールは、香料や色素として古くから食品、化粧品等に使用されてきた。1990年代以降、抗酸化作用、ホルモン促進作用等が知られるようになり、健康食品、医薬品としての適用が注目されている。 2. Polyphenol-containing composition Polyphenol is a general term for components having a plurality of phenolic hydroxy groups in the molecule. It is contained in most plants and over 5,000 types are known. Polyphenols have long been used in foods, cosmetics and the like as perfumes and pigments. Since the 1990s, antioxidative action, hormone promoting action, etc. have been known, and application as health foods and pharmaceuticals has attracted attention.
[態様1]
(1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液の上澄み液を得る
ことを含む、澄明化サトウキビジュースの製造方法。
[態様2]
豆類が、なた豆、大豆、小豆及びえんどう豆からなる群から選択される、態様1の製造方法。
[態様3]
麦類が、小麦、大麦、ライ麦及びえん麦からなる群から選択される、態様1又は2に記載の方法。
[態様4]
抽出液を添加する前のサトウキビジュースのpHを3-9の範囲に調整する工程を含む、態様1-3のいずれか1項に記載の方法。
[態様5]
抽出液が3-15重量%の濃度の固形分を含む、態様1-4のいずれか1項に記載の方法。
[態様6]
工程(1)において、サトウキビジュースの液量に対し、1-20容量%の抽出液を添加する、態様1-5のいずれか1項に記載の方法。
[態様7]
工程(2)の前に、工程(1)で得られた反応液を少なくとも30分間静置することを含む、態様1-6のいずれか1項に記載の方法。
[態様8]
石灰乳、凝集剤、あるいは、石灰乳と凝集剤の両方、をサトウキビジュースに添加する、ことを含まない、態様1-7のいずれか1項に記載の方法。
[態様9]
態様1-8のいずれか1項に記載の方法によって製造された、澄明化サトウキビジュース。
[態様10]
石灰乳に由来するカルシウムを含まない、態様9に記載の澄明化サトウキビジュース。
[態様11]
材料のサトウキビジュース中のリン酸が実質的に維持されている。態様9又は10に記載の澄明化サトウキビジュース。
[態様12]
態様9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地。
[態様13]
微生物を培養するための態様12に記載の培養用培地。
[態様14]
微生物が、酵母、乳酸菌、枯草菌、藻類からなる群から選択される、態様12の培養用培地。
[態様15]
態様9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で微生物又は細胞を培養する、ことを含む、微生物又は細胞の培養方法。
[態様16]
態様9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で微生物を培養する、ことを含む、微生物の培養方法。
[態様17]
微生物が、酵母、乳酸菌、枯草菌、藻類からなる群から選択される、態様16の方法。
[態様18]
態様9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で培養する、ことによって得られた微生物又は細胞。
[態様19]
態様9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で培養する、ことによって得られた微生物。
[態様20]
微生物が、酵母、乳酸菌、枯草菌、藻類からなる群から選択される、態様19の微生物。
[態様21]
態様19又は20の微生物より得られた微生物のエキス。
[態様22]
(1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液の上澄み液を得る、
(3)工程(2)で得られた上澄み液から糖を結晶化する、
ことを含む、製糖方法。
[態様23]
(1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液から上澄み液を除く
ことを含む、ポリフェノール含有組成物の製造方法。
[態様24]
豆類が、なた豆、大豆、小豆及びえんどう豆からなる群から選択される、態様23の製造方法。
[態様25]
麦類が、小麦、大麦、ライ麦及びえん麦からなる群から選択される、態様23又は24に記載の方法。
[態様26]
抽出液を添加する前のサトウキビジュースのpHを3-9の範囲に調整する工程を含む、態様23-25のいずれか1項に記載の方法。
[態様27]
抽出液が3-15重量%の濃度の固形分を含む、態様23-26のいずれか1項に記載の方法。
[態様28]
工程(1)において、サトウキビジュースの液量に対し、1-20容量%の抽出液を添加する、態様22-27のいずれか1項に記載の方法。
[態様29]
工程(2)の前に、工程(1)で得られた反応液を少なくとも30分間静置することを含む、態様22-28のいずれか1項に記載の方法。
[態様30]
石灰乳、凝集剤、あるいは、石灰乳と凝集剤の両方、をサトウキビジュースに添加する、ことを含まない、態様22-29のいずれか1項に記載の方法。
[態様31]
態様22-30のいずれか1項に記載の方法によって製造された、ポリフェノール含有組成物。
[態様32]
クマル酸及び/又はフェルラ酸を含む、態様31のポリフェノール含有組成物。 Although not necessarily limited, this invention includes the following aspects [Aspect 1]
(1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) A method for producing a clarified sugarcane juice, comprising obtaining a supernatant of the reaction solution in step (1).
[Aspect 2]
The production method according to
[Aspect 3]
The method according to
[Aspect 4]
The method according to any one of aspects 1-3, comprising the step of adjusting the pH of the sugarcane juice before adding the extract to a range of 3-9.
[Aspect 5]
The method according to any one of aspects 1-4, wherein the extract comprises solids at a concentration of 3-15% by weight.
[Aspect 6]
The method according to any one of aspects 1-5, wherein in step (1), 1-20% by volume of an extract is added to the amount of sugar cane juice.
[Aspect 7]
The method according to any one of Embodiments 1-6, comprising standing the reaction solution obtained in the step (1) for at least 30 minutes before the step (2).
[Aspect 8]
The method according to any one of aspects 1-7, comprising not adding lime milk, flocculant, or both lime milk and flocculant to sugarcane juice.
[Aspect 9]
A clarified sugar cane juice produced by the method according to any one of aspects 1-8.
[Aspect 10]
The clarified sugarcane juice according to aspect 9, which does not contain calcium derived from lime milk.
[Aspect 11]
The phosphate in the material sugarcane juice is substantially maintained. The clarified sugarcane juice according to
[Aspect 12]
A culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
[Aspect 13]
The culture medium according to aspect 12, for culturing microorganisms.
[Aspect 14]
The culture medium for culture according to aspect 12, wherein the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
[Aspect 15]
A method for culturing a microorganism or cell, comprising culturing the microorganism or cell in a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
[Aspect 16]
A method for culturing a microorganism, comprising culturing a microorganism in a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
[Aspect 17]
The method of
[Aspect 18]
A microorganism or cell obtained by culturing in a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
[Aspect 19]
A microorganism obtained by culturing in a culture medium comprising the clarified sugarcane juice according to any one of aspects 9-11.
[Aspect 20]
The microorganism of
[Aspect 21]
An extract of a microorganism obtained from the microorganism of
[Aspect 22]
(1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) Obtaining the supernatant of the reaction solution in step (1),
(3) Crystallize sugar from the supernatant obtained in step (2).
A sugar production method.
[Aspect 23]
(1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) A method for producing a polyphenol-containing composition, comprising removing the supernatant from the reaction liquid in step (1).
[Aspect 24]
The production method according to aspect 23, wherein the beans are selected from the group consisting of peas, soybeans, red beans, and peas.
[Aspect 25]
25. A method according to embodiment 23 or 24, wherein the wheat is selected from the group consisting of wheat, barley, rye and oats.
[Aspect 26]
The method according to any one of aspects 23-25, comprising the step of adjusting the pH of the sugarcane juice before adding the extract to a range of 3-9.
[Aspect 27]
27. A method according to any one of embodiments 23-26, wherein the extract comprises solids at a concentration of 3-15% by weight.
[Aspect 28]
28. The method according to any one of embodiments 22-27, wherein in step (1), 1-20% by volume of an extract is added to the amount of sugar cane juice.
[Aspect 29]
The method according to any one of aspects 22-28, comprising standing the reaction liquid obtained in step (1) for at least 30 minutes before step (2).
[Aspect 30]
30. A method according to any one of aspects 22-29, which does not include adding lime milk, flocculant, or both lime milk and flocculant to sugarcane juice.
[Aspect 31]
A polyphenol-containing composition produced by the method of any one of embodiments 22-30.
[Aspect 32]
The polyphenol containing composition of aspect 31 containing coumaric acid and / or ferulic acid.
本発明は一態様において、澄明化サトウキビジュースの製造方法に関する。 1. TECHNICAL FIELD The present invention, in one aspect, relates to a method for producing clarified sugar cane juice.
(1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液の上澄み液を得る
ことを含む。 Without limitation, the method for producing clarified sugarcane juice is:
(1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) including obtaining a supernatant of the reaction solution in step (1).
「サトウキビ」はイネ科サトウキビ属(Saccharum officinarum)の植物である。「サトウキビジュース」は、サトウキビから搾汁した液である。一般に、刈取りされたサトウキビの全茎又は内実部を、例えば、カッターやシュレッダー、ハンマークラッシャーにより細かく破砕した後、圧搾機によって搾汁することによって、得ることができる。 (1) Sugarcane juice “Sugar” is a plant belonging to the genus Saccharum officinarum. “Sugar cane juice” is a liquid squeezed from sugar cane. In general, the whole stem or the solid part of the cut sugar cane can be obtained by finely crushing it with, for example, a cutter, a shredder, or a hammer crusher, and then squeezing with a press.
「豆類抽出液」、「麦類抽出液」は、各々、豆類、麦類から抽出された液体である。豆類抽出液又は麦類抽出液のいずれか片方、あるいは、豆類抽出液と麦類抽出液の両方をサトウキビジュースに添加しうる。本明細書において、特に明記しない場合、「抽出液」は、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方を意味する。 (2) Extract liquid “Bean extract” and “Wheat extract” are liquids extracted from beans and wheat, respectively. Either the bean extract or the wheat extract, or both the bean extract and the wheat extract may be added to the sugar cane juice. In the present specification, unless otherwise specified, the “extract” means a bean extract, a wheat extract, or both a bean extract and a wheat extract.
澄明化サトウキビジュースの製造方法の工程(1)において、サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加する。 (3) Addition of Extract to Sugarcane Juice In step (1) of the method for producing clarified sugarcane juice, the sugarcane juice is added to the bean extract, the wheat extract, or both the bean extract and the wheat extract. Add the extract that is.
澄明化サトウキビジュースの製造方法は、工程(1)の反応液から上澄み液を得る工程を含む。工程(1)の反応液において、析出物が生じる。析出物を除くために、攪拌後、混合液から公知の固液分離法を用いて沈殿物を除去し、上澄み液を得る。このような固液分離法として、重力沈降、遠心沈降、濾過などが挙げられる。 (4) Obtaining a supernatant liquid The manufacturing method of clarified sugarcane juice includes the process of obtaining a supernatant liquid from the reaction liquid of a process (1). In the reaction solution of step (1), a precipitate is generated. In order to remove the precipitate, after stirring, the precipitate is removed from the mixed solution using a known solid-liquid separation method to obtain a supernatant. Examples of such a solid-liquid separation method include gravity sedimentation, centrifugal sedimentation, and filtration.
こうして得られた上澄み液が澄明化サトウキビジュースである。 Further, in order to increase the recovery rate of the supernatant liquid, the heavy liquid of the precipitate first separated by gravity sedimentation or centrifugal sedimentation may be filtered again, and the obtained filtrate may be mixed with the supernatant liquid.
The supernatant thus obtained is clarified sugarcane juice.
本発明は一態様において、澄明化サトウキビジュースに関する。一態様において、澄明化サトウキビジュースは、「1.澄明化サトウキビジュースの製造方法」に記載した製造方法によって製造される。 2. Clarified sugar cane juice The present invention, in one aspect, relates to clarified sugar cane juice. In one embodiment, the clarified sugarcane juice is produced by the production method described in “1. Method for producing clarified sugarcane juice”.
(i)従来のサトウキビジュースを石灰乳で処理した場合と比較して、凝集剤の使用が不要である。よって、石灰乳、凝集剤を添加するための工程が不要であり、効率的に製造できる。
(ii)サトウキビジュースを石灰乳で処理した場合と異なり、豆類等の抽出液の添加で得られた澄明化サトウキビジュースには、材料のサトウキビジュース中のリン酸が実質的に維持されている。よって、リン酸を豊富に含むため、培養用培地として使用する際、リン酸の添加量を減らすことができる。
(iii)従来の石灰乳処理したサトウキビジュース及びそれから得られる糖蜜は、カルシウムを豊富に含む。培養用培地として使用すると、カルシウムが不純物として析出してしまうため、カルシウムを除去する目的でリン酸を添加する必要が生じる。それに対し、豆類等の抽出液の添加で得られた澄明化サトウキビジュースは、リン酸を添加しなくてもよい。
(iv)豆類及び麦類の抽出液からビタミン類やミネラルが供給される。そのため、微生物等培養時にこれらの添加量を減らすことができる。 Without limitation, the clarified sugarcane juice obtained by adding an extract such as beans preferably has the following characteristics (advantages).
(I) Compared with the case where the conventional sugarcane juice is treated with lime milk, the use of a flocculant is unnecessary. Therefore, the process for adding lime milk and a flocculant is unnecessary, and it can manufacture efficiently.
(Ii) Unlike the case where sugarcane juice is treated with lime milk, the clarified sugarcane juice obtained by adding an extract such as beans substantially retains the phosphoric acid in the material sugarcane juice. Therefore, since it contains abundant phosphoric acid, the amount of phosphoric acid added can be reduced when used as a culture medium.
(Iii) Conventional lime milk-treated sugarcane juice and molasses obtained therefrom are rich in calcium. When used as a culture medium, calcium precipitates as an impurity, so that it is necessary to add phosphoric acid for the purpose of removing calcium. On the other hand, clarified sugarcane juice obtained by adding an extract such as beans does not have to be added with phosphoric acid.
(Iv) Vitamins and minerals are supplied from extracts of beans and wheat. Therefore, the amount of these added can be reduced during the cultivation of microorganisms and the like.
本発明は一態様において、「2.澄明化サトウキビジュース」に記載した澄明化サトウキビジュースを含む、培養用培地に関する。 3. Culture Medium The present invention, in one aspect, relates to a culture medium containing the clarified sugar cane juice described in “2. Clarified sugar cane juice”.
本発明は一態様において、「2.澄明化サトウキビジュース」に記載した澄明化サトウキビジュースを含む、培養用培地で培養する、ことを含む、微生物又は細胞の培養方法、に関する。 4). In one aspect, the present invention relates to a method for culturing microorganisms or cells, comprising culturing in a culture medium containing the clarified sugarcane juice described in “2. Clarified sugarcane juice”. .
本発明は、一態様において、「2.澄明化サトウキビジュース」に記載した澄明化サトウキビジュース、を含む、培養用培地中で培養する、ことによって得られた微生物又は細胞に関する。 5. Microorganism or cell In one aspect, the present invention relates to a microorganism or cell obtained by culturing in a culture medium containing the clarified sugarcane juice described in “2. Clarified sugarcane juice”.
本発明は、一態様において、本発明の微生物又は細胞より得られた微生物又は細胞のエキスに関する。 6). In one aspect, the present invention relates to a microorganism or cell extract obtained from the microorganism or cell of the present invention.
本発明は、一態様として、製糖方法に関する。製糖方法は、本発系の澄明化サトウキビジュースを製糖に利用するものである。非限定的に製糖方法は、
(1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液の上澄み液を得る、
(3)工程(2)で得られた上澄み液から糖を結晶化する、
ことを含む。 7). TECHNICAL FIELD The present invention relates to a sugar production method as one aspect. The sugar production method uses the clarified sugarcane juice of the present system for sugar production. Non-limiting sugar production methods include
(1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) Obtaining the supernatant of the reaction solution in step (1),
(3) Crystallize sugar from the supernatant obtained in step (2).
Including that.
本発明は、一態様において、ポリフェノール含有組成物の製造方法に関する。 8). TECHNICAL FIELD The present invention relates, in one aspect, to a method for producing a polyphenol-containing composition.
(1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液から上澄み液を除く
ことを含む。 Non-limiting methods for producing a polyphenol-containing composition include:
(1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) including removing the supernatant from the reaction solution of step (1).
本発明は、一態様において、ポリフェノール含有組成物に関する。 9. TECHNICAL FIELD In one aspect, the present invention relates to a polyphenol-containing composition.
乾燥したなた豆、大豆(通常の大豆及び脱脂大豆)、小豆及びえんどう豆の各種豆類、小麦及び大麦の各種麦類、並びに、玄米及びトウモロコシをミル等粉砕機で粉砕し、得られた乾燥粉砕物25gに対して60℃の蒸留水を200ml加え、攪拌機を用いて500rpmで90分間撹拌した後、3000rpmで5分間遠心分離を行った。その後、デカンテーションで上澄み液を得た。このようにして得られた上澄み液を豆類・麦類抽出液として試験に用いた。 Example 1 Preparation of Bean / Wheat Extracts and Sugarcane Juice Dried peas, soybeans (normal soybeans and defatted soybeans), various beans of red beans and peas, various wheats of wheat and barley, and Brown rice and corn are pulverized with a pulverizer such as a mill, and 200 ml of distilled water at 60 ° C. is added to 25 g of the obtained dry pulverized product, followed by stirring for 90 minutes at 500 rpm using a stirrer, followed by centrifugation at 3000 rpm for 5 minutes. went. Thereafter, a supernatant was obtained by decantation. The supernatant thus obtained was used as a legume / wheat extract in the test.
水酸化カリウム若しくはリン酸でpHを2-9に調製したサトウキビジュース45mlに実施例1で得られた各種豆類・各種麦類、玄米及びトウモロコシの各種抽出液5mlを加え、軽く撹拌した後、60℃で1時間静置した。その後、デカンテーションで上澄み液と沈殿物を分離した。 Example 2 Addition of various bean and barley extracts to sugarcane juice Various beans and various barleys obtained in Example 1 and brown rice and 45 ml of sugarcane juice prepared to pH 2-9 with potassium hydroxide or phosphoric acid After adding 5 ml of various extracts of corn and stirring gently, the mixture was allowed to stand at 60 ° C. for 1 hour. Thereafter, the supernatant and the precipitate were separated by decantation.
実施例1で得られた脱脂大豆抽出液を煮沸し、固形分が13.5%になるよう調整した。濃度調整済みの脱脂大豆抽出液を以下の表2に記載の量とり、液量が2mlとなるよう加水した。ここに、実施例1で得られたサトウキビジュースを13ml加え、軽く撹拌した後、60℃で2時間静置した。その後、遠心分離器で6000rpm、5分間処理し、デカンテーションで上澄み液と沈殿物を分離した。 Example 3 Influence of the addition amount of defatted soybean extract to sugarcane juice The defatted soybean extract obtained in Example 1 was boiled and adjusted so that the solid content was 13.5%. Concentrated defatted soybean extract was taken in the amounts shown in Table 2 below, and water was added so that the amount was 2 ml. To this, 13 ml of the sugarcane juice obtained in Example 1 was added, stirred gently, and then allowed to stand at 60 ° C. for 2 hours. Then, it processed at 6000 rpm for 5 minutes with the centrifuge, and isolate | separated the supernatant liquid and the deposit by decantation.
pH7に調製したサトウキビジュース45mlに実施例1で得られた脱脂大豆抽出液5mlを加え、軽く撹拌した後、以下の表に記載の温度で1時間静置した。その後、デカンテーションで上澄み液と沈殿物を分離した。 Example 4 Effect of temperature upon addition of various bean extracts to
また、pH5に調製したサトウキビジュース45mlに実施例1で得られた小豆抽出液 5mlを加え、軽く撹拌した後、以下の表に記載の温度で1時間静置した。その後、デカンテーションで上澄み液と沈殿物を分離した。
Further, 5 ml of the red bean extract obtained in Example 1 was added to 45 ml of sugarcane juice adjusted to
サトウキビジュース1700mlに対して、小豆抽出液(60℃、pH 7.5で抽出)180mlを添加し、室温(25℃-30℃)にて1時間撹拌の後静置することで、サトウキビジュースから湿重量として上記析出物を除去した澄明化サトウキビジュースを得た。 Example 5 Comparison of yeast culture efficiency of clarified sugarcane juice and molasses treated with beans / wheat extracts To 1700 ml of sugarcane juice, 180 ml of red bean extract (extracted at 60 ° C., pH 7.5) was added, and room temperature By stirring for 1 hour at (25 ° C.-30 ° C.), the mixture was allowed to stand to obtain a clarified sugar cane juice from which the precipitate was removed as a wet weight from the sugar cane juice.
乾燥菌体量(g):培養後の菌体を、オーブンを用いて105℃4時間の条件で乾燥させた場合の重量。
エキス固形分(g):得られた酵母エキスの乾燥重量。
エキス固形分当たりI+G(重量%):エキス固形分に含まれるイノシン酸とグアニル酸の合計重量%。高速液体クロマトグラフィー(HPLC)により測定した。
エキス中I+G総量(g):エキス固形分に含まれるイノシン酸とグアニル酸の合計量。
Dry microbial mass (g): Weight when cultivated microbial cells are dried in an oven at 105 ° C. for 4 hours.
Extract solid content (g): Dry weight of the obtained yeast extract.
I + G (% by weight) per extract solid content: Total weight% of inosinic acid and guanylic acid contained in the extract solid content. It was measured by high performance liquid chromatography (HPLC).
Total amount of I + G in extract (g): Total amount of inosinic acid and guanylic acid contained in the solid content of the extract.
実施例5の条件で培養後の、澄明化サトウキビジュース培養と糖蜜培養の培養母液について、吸光度(OD420nm)を測定した。また、培養母液中を、オーブンを用いて105℃4時間の条件で乾燥させ、得られた固形分の重量を測定した。表6の「固形分(%)」は、培養母液単位重量当たりに含まれる固形分の比率(%)である。さらに、培養母液の化学的酸素要求量(COD)を、定法(JIS K0102 17)により測定した。 Example 6 Comparison of Colors of Clarified Sugarcane Juice Treated with Beans and Wheat Extracts and Yeast Cells Cultured with Molasses Cultured mother liquors of clarified sugarcane juice culture and molasses culture after culturing under the conditions of Example 5 The absorbance (OD 420 nm) was measured. The culture mother liquor was dried using an oven at 105 ° C. for 4 hours, and the weight of the obtained solid content was measured. “Solid content (%)” in Table 6 is the ratio (%) of the solid content contained per unit weight of the culture mother liquor. Furthermore, the chemical oxygen demand (COD) of the culture mother liquor was measured by a conventional method (JIS K010217).
本実施例ではサトウキビジュースを各種豆類の抽出液を添加し、得られた析出物の成分を分析した。 Example 7 Analysis of Sugar Cane Juice Precipitate In this example, sugar bean juice was added with various bean extracts and the components of the resulting precipitate were analyzed.
カラム:InterSustain(登録商標) C18(ジーエルサイエンス株式会社)、直径2.1mmm×150mm、粒径3μm
移動相:水、アセトにトリス及び酢酸の混液
流量:0.2ml/分
カラム温度:40℃
イオン化法:エレクトロスプレー(負イオン検出モード)
設定質量数(m/z):163.0→119.0 The content of p-coumaric acid was measured as follows. First, 100 ml of methanol was added to 1 g of freeze-dried sugar cane juice precipitate in two portions, and extracted by heating and refluxing at 80 ° C. for 1 hour × 2 times. The extract was centrifuged and made up to volume with 250 ml. Subsequently, it analyzed by the liquid chromatography tandem type | mold mass spectrometry. The operating conditions of the liquid chromatography tandem mass spectrometer are as follows.
Column: InterSustain (registered trademark) C18 (GL Science Co., Ltd.), diameter 2.1 mm × 150 mm,
Mobile phase: Mixture of water, aceto with tris and acetic acid Flow rate: 0.2 ml / min Column temperature: 40 ° C
Ionization method: Electrospray (negative ion detection mode)
Set mass number (m / z): 163.0 → 119.0
カラム:COSMOSIL(登録商標) 3C18-AR-II(ナカライテスク社製)、直径4.6mm×100mm、粒径3μm
移動相:水、2-プロパノール、アセトニトリル、メタノール及び酢酸の混液
流量:1.0ml/分
カラム温度:40℃
測定波長:320nm Total ferulic acid was measured as follows. First, hydrolysis was carried out by adding 3 ml of water and 10 ml of a 1 mol / l potassium hydroxide-ethanol solution to 0.4 g of freeze-dried sugarcane juice precipitate and treating in a boiling water bath for 2 hours. To the reaction, 100 ml of water and 8 ml of 2 mol / hydrochloric acid were added. To this, 50 ml of ethyl acetate was added three times to dissolve in the ethyl acetate layer. After dehydrating and drying, 10 ml of methanol was added. Next, the operating conditions of the high performance liquid chromatography analyzed by high performance liquid chromatography are as follows.
Column: COSMOSIL (registered trademark) 3C 18 -AR-II (manufactured by Nacalai Tesque), diameter 4.6 mm × 100 mm,
Mobile phase: Water, 2-propanol, acetonitrile, methanol and acetic acid mixture Flow rate: 1.0 ml / min Column temperature: 40 ° C
Measurement wavelength: 320 nm
図3に示したフローチャートに基づき、サトウキビジュースに小豆抽出物を添加し、上澄み液及び沈殿物中のポリフェノールの含量を調べた。ポリフェノールの含量は、実施例7と同様に、フォーリン-チオカルト法を用いて測定し、没食子酸の検量線を標準として換算した。
BSL 15:黒砂糖を15%w/vに溶解した砂糖溶液。
黒砂糖の調整法 サトウキビジュースを煮詰めて、発生した澱を除去しながら濃縮する。水の蒸発がなくなる5倍濃縮で煮詰め作業を終了する。種晶が形成するように白みを帯びるまで充分攪拌する。一晩冷却して固化した黒砂糖を得る。
RBE 70:小豆を70℃蒸留水で30分処理した抽出物
TPP:ポリフェノール総量
湿潤残渣(wet residue)中のポリフェノール総量は0.04182gで、湿潤残渣中へのポリフェノール回収率は36.5%であった。 Example 8 Analysis of polyphenols in the supernatant and precipitate of sugarcane juice to which the red bean extract was added Based on the flowchart shown in FIG. 3, the red bean extract was added to the sugarcane juice and the polyphenols in the supernatant and precipitate were The content was examined. The content of polyphenol was measured using the foreign-thiocult method in the same manner as in Example 7 and converted using a calibration curve of gallic acid as a standard.
BSL 15: A sugar solution in which brown sugar is dissolved in 15% w / v.
How to prepare brown sugar Boil sugar cane juice and concentrate while removing the generated starch. The simmering operation is completed with a 5-fold concentration that eliminates water evaporation. Stir well until it turns white so that seed crystals form. Cool overnight to get solidified brown sugar.
RBE 70: Extract obtained by treating red beans with distilled water at 70 ° C. for 30 minutes TPP: Total amount of polyphenol The total amount of polyphenol in the wet residue was 0.04182 g, and the polyphenol recovery rate in the wet residue was 36.5% there were.
図4又は図5に示したフローチャートに基づき、サトウキビジュースに脱脂大豆抽出物を添加し、上澄み液及び沈殿物中のポリフェノールの含量を調べた。 Example 9 Analysis of polyphenol in the supernatant and precipitate of sugarcane juice to which defatted soybean extract was added Based on the flowchart shown in FIG. 4 or FIG. 5, the defatted soybean extract was added to sugarcane juice, and the supernatant and precipitate were added. The content of polyphenol in the product was examined.
図4、図5に示したように2回の試験を行った。1回目は、湿潤残渣(wet residue)中のポリフェノール総量は0.046gで、湿潤残渣中へのポリフェノール回収率は32.81%であった。2回目は、湿潤残渣(wet residue)中のポリフェノール総量は0.021gで、湿潤残渣中へのポリフェノール回収率は15.00%であった。 DFSBE 70: Extract obtained by treating defatted soybeans with distilled water at 70 ° C. for 30 minutes As shown in FIGS. 4 and 5, two tests were performed. In the first round, the total amount of polyphenols in the wet residue was 0.046 g, and the polyphenol recovery rate in the wet residue was 32.81%. The second time, the total amount of polyphenols in the wet residue was 0.021 g, and the polyphenol recovery rate in the wet residue was 15.00%.
本実施例では、脱脂大豆抽出物を添加したサトウキビジュースから得られた沈殿物を用いて、DPPHラジカル消去活性により抗酸化活性を測定した。 Example 9 Measurement of Antioxidant Activity of Precipitates Obtained from Sugarcane Juice In this example, antioxidant activity was measured by DPPH radical scavenging activity using precipitates obtained from sugarcane juice to which defatted soybean extract was added. did.
400μMのDPPH溶液は、先ずDPPHをエタノールで十分に溶解してから(0.5時間-1時間)、蒸留水を加えた。
In the 400 μM DPPH solution, DPPH was first sufficiently dissolved with ethanol (0.5 hour-1 hour), and distilled water was added thereto.
本実施例では、サトウキビジュースから得られた沈殿物を含む溶液の黄色の濃さを示す420nmの吸光度を調べた。サトウキビジュースから得られた沈殿物として、小豆抽出物を添加したサトウキビジュースの沈殿物(RBE)及び脱脂大豆抽出物を添加したサトウキビジュースの沈殿物(DFSBE)を用いた。対照として、MSX-100(三井製糖)及びMSX-1MF(三井製糖)を用いた。いずれのサンプルも1%可溶性固形分 溶液について測定した。 Example 10 Absorbance of a Solution Containing a Precipitate Obtained from Sugarcane Juice In this example, an absorbance at 420 nm indicating the yellowness of a solution containing a precipitate obtained from sugarcane juice was examined. As a precipitate obtained from sugarcane juice, a sugarcane juice precipitate (RBE) to which an adzuki bean extract was added and a sugarcane juice precipitate (DFSBE) to which a defatted soybean extract was added were used. As controls, MSX-100 (Mitsui Sugar) and MSX-1MF (Mitsui Sugar) were used. All samples were measured on a 1% soluble solids solution.
Claims (32)
- (1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液の上澄み液を得る
ことを含む、澄明化サトウキビジュースの製造方法。 (1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) A method for producing a clarified sugarcane juice, comprising obtaining a supernatant of the reaction solution in step (1). - 豆類が、なた豆、大豆、小豆及びえんどう豆からなる群から選択される、請求項1の製造方法。 The method according to claim 1, wherein the beans are selected from the group consisting of peas, soybeans, red beans, and peas.
- 麦類が、小麦、大麦、ライ麦及びえん麦からなる群から選択される、請求項1又は2に記載の方法。 The method according to claim 1 or 2, wherein the wheat is selected from the group consisting of wheat, barley, rye and oats.
- 抽出液を添加する前のサトウキビジュースのpHを3-9の範囲に調整する工程を含む、請求項1-3のいずれか1項に記載の方法。 The method according to any one of claims 1 to 3, comprising a step of adjusting the pH of sugarcane juice before adding the extract to a range of 3-9.
- 抽出液が3-15重量%の濃度の固形分を含む、請求項1-4のいずれか1項に記載の方法。 The method according to any one of claims 1 to 4, wherein the extract contains a solid content of 3 to 15% by weight.
- 工程(1)において、サトウキビジュースの液量に対し、1-20容量%の抽出液を添加する、請求項1-5のいずれか1項に記載の方法。 The method according to any one of claims 1 to 5, wherein in step (1), 1 to 20% by volume of an extract is added to the amount of sugar cane juice.
- 工程(2)の前に、工程(1)で得られた反応液を少なくとも30分間静置することを含む、請求項1-6のいずれか1項に記載の方法。 The method according to any one of claims 1 to 6, comprising standing the reaction solution obtained in step (1) for at least 30 minutes before step (2).
- 石灰乳、凝集剤、あるいは、石灰乳と凝集剤の両方、をサトウキビジュースに添加する、ことを含まない、請求項1-7のいずれか1項に記載の方法。 The method according to any one of claims 1 to 7, which does not include adding lime milk, flocculant, or both lime milk and flocculant to sugarcane juice.
- 請求項1-8のいずれか1項に記載の方法によって製造された、澄明化サトウキビジュース。 A clarified sugarcane juice produced by the method according to any one of claims 1-8.
- 石灰乳に由来するカルシウムを含まない、請求項9に記載の澄明化サトウキビジュース。 The clarified sugarcane juice according to claim 9, which does not contain calcium derived from lime milk.
- 材料のサトウキビジュース中のリン酸が実質的に維持されている。請求項9又は10に記載の澄明化サトウキビジュース。 The phosphoric acid in the material sugarcane juice is substantially maintained. The clarified sugarcane juice according to claim 9 or 10.
- 請求項9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地。 A culture medium comprising the clarified sugarcane juice according to any one of claims 9-11.
- 微生物を培養するための請求項12に記載の培養用培地。 The culture medium according to claim 12 for culturing microorganisms.
- 微生物が、酵母、乳酸菌、枯草菌、藻類からなる群から選択される、請求項12の培養用培地。 The culture medium according to claim 12, wherein the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- 請求項9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で微生物又は細胞を培養する、ことを含む、微生物又は細胞の培養方法。 A method for culturing microorganisms or cells, comprising culturing microorganisms or cells in a culture medium containing the clarified sugarcane juice according to any one of claims 9-11.
- 請求項9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で微生物を培養する、ことを含む、微生物の培養方法。 A method for culturing a microorganism, comprising culturing a microorganism in a culture medium containing the clarified sugarcane juice according to any one of claims 9-11.
- 微生物が、酵母、乳酸菌、枯草菌、藻類からなる群から選択される、請求項16の方法。 The method according to claim 16, wherein the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- 請求項9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で培養する、ことによって得られた微生物又は細胞。 A microorganism or cell obtained by culturing in a culture medium containing the clarified sugarcane juice according to any one of claims 9-11.
- 請求項9-11のいずれか1項に記載の澄明化サトウキビジュース、を含む、培養用培地中で培養する、ことによって得られた微生物。 A microorganism obtained by culturing in a culture medium containing the clarified sugarcane juice according to any one of claims 9-11.
- 微生物が、酵母、乳酸菌、枯草菌、藻類からなる群から選択される、請求項19の微生物。 The microorganism according to claim 19, wherein the microorganism is selected from the group consisting of yeast, lactic acid bacteria, Bacillus subtilis, and algae.
- 請求項19又は20の微生物より得られた微生物のエキス。 A microorganism extract obtained from the microorganism according to claim 19 or 20.
- (1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液の上澄み液を得る、
(3)工程(2)で得られた上澄み液から糖を結晶化する、
ことを含む、製糖方法。 (1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) Obtaining the supernatant of the reaction solution in step (1),
(3) Crystallize sugar from the supernatant obtained in step (2).
A sugar production method. - (1)サトウキビジュースに、豆類抽出液、麦類抽出液、あるいは、豆類抽出液と麦類抽出液の両方、である抽出液を添加し、
(2)工程(1)の反応液から上澄み液を除く
ことを含む、ポリフェノール含有組成物の製造方法。 (1) To sugarcane juice, add an extract that is a bean extract, a wheat extract, or both a bean extract and a wheat extract,
(2) A method for producing a polyphenol-containing composition, comprising removing the supernatant from the reaction liquid in step (1). - 豆類が、なた豆、大豆、小豆及びえんどう豆からなる群から選択される、請求項23の製造方法。 The method according to claim 23, wherein the beans are selected from the group consisting of peas, soybeans, red beans, and peas.
- 麦類が、小麦、大麦、ライ麦及びえん麦からなる群から選択される、請求項23又は24に記載の方法。 25. The method according to claim 23 or 24, wherein the wheat is selected from the group consisting of wheat, barley, rye and oats.
- 抽出液を添加する前のサトウキビジュースのpHを3-9の範囲に調整する工程を含む、請求項23-25のいずれか1項に記載の方法。 The method according to any one of claims 23 to 25, comprising a step of adjusting the pH of the sugarcane juice before adding the extract to a range of 3-9.
- 抽出液が3-15重量%の濃度の固形分を含む、請求項23-26のいずれか1項に記載の方法。 The method according to any one of claims 23 to 26, wherein the extract contains a solid content of 3 to 15% by weight.
- 工程(1)において、サトウキビジュースの液量に対し、1-20容量%の抽出液を添加する、請求項22-27のいずれか1項に記載の方法。 The method according to any one of claims 22 to 27, wherein in the step (1), 1 to 20% by volume of an extract is added to the amount of sugar cane juice.
- 工程(2)の前に、工程(1)で得られた反応液を少なくとも30分間静置することを含む、請求項22-28のいずれか1項に記載の方法。 The method according to any one of claims 22 to 28, comprising standing the reaction liquid obtained in the step (1) for at least 30 minutes before the step (2).
- 石灰乳、凝集剤、あるいは、石灰乳と凝集剤の両方、をサトウキビジュースに添加する、ことを含まない、請求項22-29のいずれか1項に記載の方法。 30. A method according to any one of claims 22-29, which does not comprise adding lime milk, flocculant, or both lime milk and flocculant to sugarcane juice.
- 請求項22-30のいずれか1項に記載の方法によって製造された、ポリフェノール含有組成物。 A polyphenol-containing composition produced by the method according to any one of claims 22-30.
- クマル酸及び/又はフェルラ酸を含む、請求項31のポリフェノール含有組成物。 32. The polyphenol-containing composition of claim 31, comprising coumaric acid and / or ferulic acid.
Priority Applications (3)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
BR112020016100-4A BR112020016100A2 (en) | 2018-02-28 | 2018-11-30 | CLARIFIED SUGARCANE SUGAR AND COMPOSITION CONTAINING POLYPHENOL |
CN201880090379.6A CN111936615A (en) | 2018-02-28 | 2018-11-30 | Clarified sugarcane juice and polyphenol-containing composition |
JP2020502811A JP7252193B2 (en) | 2018-02-28 | 2018-11-30 | Clarified sugarcane juice and polyphenol-containing composition |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2018035543 | 2018-02-28 | ||
JP2018-035543 | 2018-02-28 |
Publications (1)
Publication Number | Publication Date |
---|---|
WO2019167371A1 true WO2019167371A1 (en) | 2019-09-06 |
Family
ID=67805801
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/JP2018/044131 WO2019167371A1 (en) | 2018-02-28 | 2018-11-30 | Clarified sugar cane juice and polyphenol-containing composition |
Country Status (4)
Country | Link |
---|---|
JP (1) | JP7252193B2 (en) |
CN (1) | CN111936615A (en) |
BR (1) | BR112020016100A2 (en) |
WO (1) | WO2019167371A1 (en) |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016175235A1 (en) * | 2015-04-28 | 2016-11-03 | テーブルマーク株式会社 | Method for producing yeast extract, yeast extract obtained thereby, seasoning composition and food |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2010503417A (en) * | 2006-09-19 | 2010-02-04 | ホリズン サイエンス ピーティーワイ リミテッド | Extracts derived from sugarcane and methods for producing them |
CN105755180B (en) * | 2016-03-14 | 2019-05-10 | 广西大学 | The defecation method of sugar-cane juice |
-
2018
- 2018-11-30 WO PCT/JP2018/044131 patent/WO2019167371A1/en active Application Filing
- 2018-11-30 BR BR112020016100-4A patent/BR112020016100A2/en unknown
- 2018-11-30 JP JP2020502811A patent/JP7252193B2/en active Active
- 2018-11-30 CN CN201880090379.6A patent/CN111936615A/en active Pending
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2016175235A1 (en) * | 2015-04-28 | 2016-11-03 | テーブルマーク株式会社 | Method for producing yeast extract, yeast extract obtained thereby, seasoning composition and food |
Non-Patent Citations (2)
Title |
---|
KUMAR P.: "New approaches for jaggery production in Rajasthan", IND. J. SUGARCANE TECHNOL., vol. 30, no. 1, 2015, pages 6 - 11 * |
MUNGARE T.S. ET AL.: "Clarification technique for producing quality jaggery", COOPERATIVE SUGAR, vol. 32, no. 4, 2000, pages 283 - 285 * |
Also Published As
Publication number | Publication date |
---|---|
BR112020016100A2 (en) | 2020-12-08 |
CN111936615A (en) | 2020-11-13 |
JPWO2019167371A1 (en) | 2021-02-12 |
JP7252193B2 (en) | 2023-04-04 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
AU2004276123B2 (en) | Fermentation and culture method, fermented plant extract, fermented plant extract powder and composition containing the fermented plant extract | |
KR101025286B1 (en) | Method for Preparing Functional Natural Fermentation Condiment and Sea-Tangle Fermentation Powder Containing GABA Using Fermentation of Sea-Tangle | |
KR101377586B1 (en) | Method for producing fermented inonotus obliquus with increased antioxidant activity and total phenolic compounds by the solid-state fermentation | |
WO2016204120A1 (en) | Method for producing fermented bamboo extract and immunostimulator | |
KR101368688B1 (en) | A preparation method of rubus coreanus fruit wine for producing non-alcoholic fermentation product of rubus coreanus fruits | |
JP4671450B1 (en) | Diet food | |
KR101661495B1 (en) | The method of Eco-friendly alcohol-free, water-soluble propolis using natural honey | |
JPWO2019065396A1 (en) | Production method of purified Salacia plant extract and purified Salacia plant extract | |
Kartawiria et al. | Sorghum stalk juice pre-treatment method for bioethanol fermentation process | |
WO2009024316A1 (en) | Method of removal of bitter taste from olive juice extract | |
WO2019167371A1 (en) | Clarified sugar cane juice and polyphenol-containing composition | |
Upadhyaya et al. | Palmyrah palm and its products (Neera, Jaggery and Candy)—A Review on chemistry and technology | |
RU2613294C1 (en) | Method for producing melanin from sunflower husks | |
KR20140086901A (en) | Functional food for improving the treatment of diabetes and obesity comprising Momordica charantia, chounggukjang and sea tangle | |
KR20060060601A (en) | Process for preparing a fermentation food containing chlorella | |
KR101672098B1 (en) | Composition for removing hangover comprising fermented rice rinse water | |
KR102073303B1 (en) | Manufacture of scorched-rice water using germinated/fermented brown rice with high content of GABA | |
KR20130120562A (en) | Method for lentinula edodes extracts and the extract | |
Saparbekova | Extraction of resveratrol from the pomace of various grape varieties using an enzyme preparation | |
KR101060514B1 (en) | Functional natural fermented seasonings and kelp fermented powder from kelp fermentation products by yeast and preparation method thereof | |
KR101957573B1 (en) | Environmentally friendly feed for livestock using bokbunja byproducts and its preparation method | |
Mahmood et al. | Evaluation of physical and chemical properties and total phenolic content in baker’s yeast obtained from grape juice | |
JP2009044965A (en) | Sugarcane functional extract and method for producing the same | |
KR100392775B1 (en) | Alismatis Rhizoma tea of rosted type and process for prepatration thereof | |
CN106520873A (en) | Preparation method for peptide chelated calcium of clanis bilineata tsingtauica larva |
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: 18907653 Country of ref document: EP Kind code of ref document: A1 |
|
DPE1 | Request for preliminary examination filed after expiration of 19th month from priority date (pct application filed from 20040101) | ||
ENP | Entry into the national phase |
Ref document number: 2020502811 Country of ref document: JP Kind code of ref document: A |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
REG | Reference to national code |
Ref country code: BR Ref legal event code: B01A Ref document number: 112020016100 Country of ref document: BR |
|
ENP | Entry into the national phase |
Ref document number: 112020016100 Country of ref document: BR Kind code of ref document: A2 Effective date: 20200806 |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 18907653 Country of ref document: EP Kind code of ref document: A1 |