JP6995206B2 - How to increase the yield of extracellular polysaccharides of edible and medicinal fungi by adding phenylalanine - Google Patents
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Description
本発明は、フェニルアラニンの添加により食用・薬用真菌の細胞外多糖の収量を高める方法に関し、微生物発酵分野に属する。 The present invention belongs to the field of microbial fermentation with respect to a method for increasing the yield of extracellular polysaccharides of edible and medicinal fungi by adding phenylalanine.
食用菌は重要な食料源の１つであり、多種のビタミン、ミネラルおよび食物繊維に富み、高タンパク質、低脂肪、低カロリーで、老若に適している食品であり、また、２１世紀の人類食料ではめったにないグリーンフードでもある。食用・薬用真菌に含まれる真菌多糖、ポリペプチド類及びその他の極めて豊富な生物活性成分には、抗腫瘍、血中脂質低下、生体の免疫調節、心臓・脳血管保護など、さまざまな薬用価値がある。たとえば、霊芝（Ｇａｎｏｄｅｒｍａ ｌｕｃｉｄｕｍ）は、優れた栄養機能及び薬用価値を備えた貴重な食用・薬用真菌である。霊芝多糖は霊芝の主要な生物活性物質の一つであり、免疫調節、抗腫瘍、抗酸化、血糖低下、肝臓保護などの活性を有する。これらの生物活性は、霊芝多糖が食品、医薬及び化粧品の分野に巨大な応用可能性を有することになる。 Edible fungi are one of the important food sources, rich in various vitamins, minerals and dietary fiber, high in protein, low in fat, low in calories, suitable for young and old, and human food in the 21st century. It's also a rare green food. Fungal polysaccharides, polypeptides and other extremely abundant bioactive ingredients contained in edible and medicinal fungi have various medicinal values such as antitumor, blood lipid lowering, biological immunomodulation, and cardiovascular protection. be. For example, Ganoderma lucidum is a valuable edible and medicinal fungus with excellent nutritional function and medicinal value. Reishi polysaccharide is one of the major bioactive substances of Reishi, and has activities such as immunomodulation, antitumor, antioxidant, hypoglycemic, and liver protection. These biological activities will allow Reishi polysaccharides to have enormous potential in the fields of food, medicine and cosmetics.
現在、液体深部発酵技術による食用・薬用真菌の培養は、食用・薬用の真菌多糖を得るための主な発酵方式となっており、固相発酵に比べて発酵サイクルが短く、抽出コストが低くなるものの、多糖の収量は比較的低く、食用・薬用真菌多糖の発酵収量を如何に大幅に増加させて製造コストをさらに低減し、上記分野での幅広い用途を達成することは、食用・薬用真菌の液体発酵において解決すべき急務となっている。 Currently, culturing edible and medicinal fungi using deep liquid fermentation technology is the main fermentation method for obtaining edible and medicinal fungal polysaccharides, and the fermentation cycle is shorter and the extraction cost is lower than solid-phase fermentation. However, the yield of polysaccharides is relatively low, and how to significantly increase the fermentation yield of edible and medicinal fungi polysaccharides to further reduce manufacturing costs and achieve a wide range of applications in the above fields is the ability of edible and medicinal fungi. There is an urgent need to solve this in liquid fermentation.
本発明の第１目的は、真菌発酵の初期又は発酵中にフェニルアラニンを添加する真菌細胞外多糖の収量を高める方法を提供することである。 A first object of the present invention is to provide a method for increasing the yield of fungal extracellular polysaccharides to which phenylalanine is added in the early stage of fungal fermentation or during fermentation.
本発明の一実施形態では、前記真菌は食用・薬用真菌である。 In one embodiment of the invention, the fungus is an edible / medicinal fungus.
本発明の一実施形態では、前記食用・薬用真菌には、霊芝、キクラゲ、シイタケ、チョレイマイタケ、シロキクラゲ、マイタケ、マツホド、カワラタケ、ヤマブシタケ、冬虫夏草及び通常の生物分類学的に近縁種である又は形質が近い食用・薬用真菌の子実体又は菌糸体が含まれるが、これらに制限されない。 In one embodiment of the present invention, the edible and medicinal fungi include Reishi, Kikurage, Shiitake, Choreimaitake, White fungus, Maitake, Matsuhodo, Kawaratake, Yamabushitake, Fuyu-mushi summer grass and ordinary bioclassical relatives. Includes, but is not limited to, fruiting bodies or mycelia of edible / medicinal fungi with or with similar traits.
本発明の一実施形態では、前記真菌には、霊芝（Ｇａｎｏｄｅｒｍａ ｌｕｃｉｄｕｍ）、アギタケ（Ｐｌｅｕｒｏｔｕｓ ｆｅｒｕｌａｅ）、アラゲカワラタケ（Ｃｏｒｉｏｌｕｓｈｉｓｕｔｕｓ）、スエヒロタケ（Ｓｃｈｉｚｏｐｈｙｌｌｕｍｃｏｍｍｕｎｅ）が含まれるが、これらに制限されない。 In one embodiment of the invention, the fungus includes, but is not limited to, Ganoderma lucidum, Pleurotus ferulae, Coriolus siustus, and Schizophyllum commune.
本発明の一実施形態では、前記方法において、発酵の０～９６ｈでフェニルアラニンを添加する。 In one embodiment of the invention, in the above method, phenylalanine is added from 0 to 96 hours of fermentation.
本発明の一実施形態では、前記フェニルアラニンの最終濃度が０．０５～３．５ｇ／Ｌである。 In one embodiment of the invention, the final concentration of phenylalanine is 0.05-3.5 g / L.
本発明の一実施形態では、前記方法では、真菌を発酵培地に接種して、発酵培地にフェニルアラニンを添加する。 In one embodiment of the invention, in the method, the fungus is inoculated into the fermentation medium and phenylalanine is added to the fermentation medium.
本発明の一実施形態では、発酵培地は、食用・薬用真菌によく使用される液体発酵培地である。 In one embodiment of the invention, the fermentation medium is a liquid fermentation medium often used for edible and medicinal fungi.
本発明の一実施形態では、前記液体発酵培地１Ｌあたり、グルコース２０ｇ、トリプトン５ｇ、アミノ酸不含酵母（ＹＮＢ）５ｇ、リン酸二水素カリウム４．５ｇ、硫酸マグネシウム七水和物２ｇが含まれ、この培地は自然ｐＨである。 In one embodiment of the present invention, 20 g of glucose, 5 g of tryptone, 5 g of amino acid-free yeast (YNB), 4.5 g of potassium dihydrogen phosphate, and 2 g of magnesium sulfate heptahydrate are contained in 1 L of the liquid fermentation medium. This medium has a natural pH.
本発明の第２目的は、真菌細胞外多糖の製造方法を提供し、前記方法では、真菌を発酵培地に接種し、真菌発酵の初期又は発酵中に発酵培地へフェニルアラニンを添加し、２５～３３℃、１５０～２００ｒ・ｍｉｎ－１で５～７日間、発酵させる。 A second object of the present invention is to provide a method for producing a fungal extracellular polysaccharide, in which the fungus is inoculated into a fermentation medium and phenylalanine is added to the fermentation medium in the early stage or during fermentation of the fungus, 25-33. Ferment at 150-200 r · min -1 at ° C for 5-7 days.
本発明の一実施形態では、３～６ｇ菌糸体湿重量／Ｌ培地の接種量で接種する。 In one embodiment of the present invention, the inoculation amount is 3 to 6 g of mycelium wet weight / L medium.
本発明は、細胞外多糖を含む製品を製造するための、前記方法の使用をさらに提供する。 The present invention further provides the use of said method for producing products comprising extracellular polysaccharides.
有益な効果は以下のとおりである。本発明では、発酵中にフェニルアラニンを添加することにより、元の発酵サイクルを延ばすことなく、食用・薬用真菌の細胞外多糖の収量を著しく増加させ、最大では４０％以上向上させ、食用・薬用真菌多糖の製造コストを大幅に低減させ、工業的生産や製品の応用に寄与する。 The beneficial effects are: In the present invention, the addition of phenylalanine during fermentation significantly increases the yield of extracellular polysaccharides of edible and medicinal fungi without prolonging the original fermentation cycle, improving by up to 40% or more, and edible and medicinal fungi. It significantly reduces the cost of producing polysaccharides and contributes to industrial production and product application.
多糖の抽出
発酵液のろ液を１００ｍＬ採取し、４倍の９５％アルコールを加え、２０ｍｉｎ撹拌して、４０００ｒ・ｍｉｎ－１で５ｍｉｎ遠心分離し、タンパク質を除去して、上澄み液に２．２５倍の９５％アルコールを加え、２０ｍｉｎ撹拌後、４℃で一晩静置した。溶液を１００００ｒ・ｍｉｎ－１で５ｍｉｎ遠心分離し、上澄みを除去して、沈殿に蒸留水３０ｍＬを加えて振とうして溶解させ、１００００ｒ・ｍｉｎ－１で１０ｍｉｎ遠心分離し、透明液として水溶性多糖溶液を得た。
多糖の測定
多糖の含有量測定には、フェノール硫酸法が使用され、測定系は、サンプル溶液２ｍＬ、６％フェノール１ｍＬ、濃硫酸５ｍＬである。反応終了後、冷却し、その後、４９０ｎｍ波長でＯＤ値を測定した。
Extraction of polysaccharides Collect 100 mL of the filtrate of the fermentation broth, add 4 times 95% alcohol, stir for 20 min, centrifuge at 4000 r · min -1 for 5 min, remove the protein, and add 2.25 to the supernatant. Twice as much 95% alcohol was added, and the mixture was stirred for 20 minutes and then allowed to stand at 4 ° C. overnight. Centrifuge the solution at 10000 r · min -1 for 5 min, remove the supernatant, add 30 mL of distilled water to the precipitate, shake to dissolve, centrifuge at 10000 r · min -1 for 10 min, and water-soluble as a clear solution. A polysaccharide solution was obtained.
Measurement of polysaccharide The phenol-sulfuric acid method is used to measure the content of polysaccharide, and the measurement system is 2 mL of sample solution, 1 mL of 6% phenol, and 5 mL of concentrated sulfuric acid. After completion of the reaction, the mixture was cooled, and then the OD value was measured at a wavelength of 490 nm.
実施例１
霊芝のシード培養
サイズ０．５ｃｍ２の菌塊を採取し、シード培地の液体仕込み量が８０ｍＬ／２５０ｍＬの三角フラスコに接種して、１５０ｒ・ｍｉｎ－１、３０℃で７日間、培養した。
霊芝の発酵培養
５００ｍＬ三角フラスコに発酵培地１５０ｍＬを加え、１１５℃で２０分殺菌した。発酵培地に湿重量０．５ｇの霊芝菌糸体を接種して、１５０ｒ・ｍｉｎ－１、３０℃で７日間、培養した。
前記シード培地と発酵培地の配合は以下のとおりである。１Ｌあたり、グルコース２０ｇ、トリプトン５ｇ、無アミノ基酵母（ＹＮＢ）５ｇ、リン酸二水素カリウム４．５ｇ、硫酸マグネシウム七水和物２ｇが含まれ、この培地は自然ｐＨである。
発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、７日間培養した霊芝多糖の収量は０．３２０ｇ／Ｌ発酵上澄み液であった。
Example 1
Reishi seed culture A bacterial mass having a size of 0.5 cm 2 was collected, inoculated into an Erlenmeyer flask having a liquid charge of 80 mL / 250 mL, and cultured at 150 r · min -1 , 30 ° C. for 7 days.
Fermentation culture of Reishi A 150 mL of fermentation medium was added to a 500 mL Erlenmeyer flask and sterilized at 115 ° C. for 20 minutes. The fermentation medium was inoculated with Reishi mycelium having a wet weight of 0.5 g and cultured at 150 r · min -1 at 30 ° C. for 7 days.
The composition of the seed medium and the fermentation medium is as follows. Each L contains 20 g of glucose, 5 g of tryptone, 5 g of amino-free yeast (YNB), 4.5 g of potassium dihydrogen phosphate, and 2 g of magnesium sulfate heptahydrate, and this medium has a natural pH.
During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide cultured for 7 days was 0.320 g / L fermentation supernatant.
実施例２
０ｈに発酵培地へフェニルアラニンを培地中のフェニルアラニンの最終濃度が０．０５ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１と同様であった。発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、霊芝多糖の収量は０．４００ｇ／Ｌ発酵上澄み液であり、実施例１よりも２５％向上した。
Example 2
The medium and culture method were the same as in Example 1 except that phenylalanine was added to the fermentation medium at 0 h so that the final concentration of phenylalanine in the medium was 0.05 g / L. During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide was 0.400 g / L fermentation supernatant, which was 25% higher than that of Example 1.
実施例３
０ｈに発酵培地へフェニルアラニンをその最終濃度が３．５ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１と同様であった。発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、霊芝多糖の収量は０．３８４ｇ／Ｌ発酵上澄み液であり、実施例１よりも２０％向上した。
Example 3
The medium and culture method were the same as in Example 1 except that phenylalanine was added to the fermentation medium at 0 h so that the final concentration was 3.5 g / L. During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide was 0.384 g / L fermentation supernatant, which was 20% higher than that of Example 1.
実施例４
０ｈに発酵培地へフェニルアラニンをその最終濃度が１ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１と同様であった。発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、霊芝多糖の収量は０．４３２ｇ／Ｌ発酵上澄み液であり、実施例１よりも３５％向上した。
Example 4
The medium and culture method were the same as in Example 1 except that phenylalanine was added to the fermentation medium at 0 h so that the final concentration was 1 g / L. During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide was 0.432 g / L fermentation supernatant, which was 35% higher than that of Example 1.
実施例５
４８ｈの発酵でフェニルアラニンをその最終濃度が２ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１と同様であった。発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、霊芝多糖の収量は０．４４８ｇ／Ｌ発酵上澄み液であり、実施例１よりも４０％向上した。
Example 5
The medium and culture method were the same as in Example 1 except that phenylalanine was added so as to have a final concentration of 2 g / L in the fermentation for 48 hours. During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide was 0.448 g / L fermentation supernatant, which was 40% higher than that of Example 1.
実施例６
９６ｈの発酵でフェニルアラニンをその最終濃度が１ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１と同様であった。発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、霊芝多糖の収量は０．４１６ｇ／Ｌ発酵上澄み液であり、実施例１よりも３０％向上した。
Example 6
The medium and culture method were the same as in Example 1 except that phenylalanine was added so as to have a final concentration of 1 g / L in the fermentation for 96 hours. During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide was 0.416 g / L fermentation supernatant, which was 30% higher than that of Example 1.
実施例７
霊芝のシード培養
サイズ０．５ｃｍ２の菌塊を採取し、シード培地の液体仕込み量が８０ｍＬ／２５０ｍＬの三角フラスコに接種して、１５０ｒ・ｍｉｎ－１、３０℃で７日間、培養した。
霊芝の発酵培養
５００ｍＬの三角フラスコに発酵培地１５０ｍＬを加え、１１５℃で２０分殺菌した。発酵培地に湿重量０．５ｇの霊芝菌糸体を接種して、１５０ｒ・ｍｉｎ－１、３０℃で７日間、培養した。
前記シード培地と発酵培地の配合は以下のとおりである。１Ｌあたり、グルコース２０ｇ、トウモロコシ粉１０ｇ、ふすま１０ｇ、リン酸二水素カリウム３ｇ、硫酸マグネシウム七水和物２ｇが含まれ、この培地は自然ｐＨである。
発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、７日間、発酵させたとき、霊芝多糖の収量は０．３６８ｇ／Ｌ発酵上澄み液であった。
Example 7
Reishi seed culture A bacterial mass having a size of 0.5 cm 2 was collected, inoculated into an Erlenmeyer flask having a liquid charge of 80 mL / 250 mL, and cultured at 150 r · min -1 , 30 ° C. for 7 days.
Fermentation culture of Reishi A 150 mL of fermentation medium was added to a 500 mL Erlenmeyer flask and sterilized at 115 ° C. for 20 minutes. The fermentation medium was inoculated with Reishi mycelium having a wet weight of 0.5 g and cultured at 150 r · min -1 at 30 ° C. for 7 days.
The composition of the seed medium and the fermentation medium is as follows. Each liter contains 20 g of glucose, 10 g of corn flour, 10 g of bran, 3 g of potassium dihydrogen phosphate and 2 g of magnesium sulfate heptahydrate, and this medium has a natural pH.
During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, when fermented for 7 days, the yield of Reishi polysaccharide was 0.368 g / L fermented supernatant.
実施例８
０ｈに発酵培地へフェニルアラニンをその最終濃度が１ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例７と同様であった。発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、霊芝多糖の収量は０．４４２ｇ／Ｌ発酵上澄み液であり、実施例７よりも２０％向上した。
Example 8
The medium and culture method were the same as in Example 7 except that phenylalanine was added to the fermentation medium at 0 h so that the final concentration was 1 g / L. During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide was 0.442 g / L fermentation supernatant, which was 20% higher than that of Example 7.
実施例９
霊芝のシード培養
サイズ０．５ｃｍ２の菌塊を採取し、シード培地の液体仕込み量が８０ｍＬ／２５０ｍＬの三角フラスコに接種して、１５０ｒ・ｍｉｎ－１、３０℃で７日間、培養した。
霊芝の発酵培養
５Ｌ発酵タンクに発酵培地３Ｌを加え、１１５℃で２０分殺菌した。湿重量１０ｇの霊芝菌糸体を接種し、１５０ｒ・ｍｉｎ－１、通気量１．５Ｌ・ｍｉｎ－１、３０℃で７日間、培養した。
前記シード培地と発酵培地の配合は以下のとおりである。１Ｌあたり、グルコース２０ｇ、トウモロコシ粉１０ｇ、ふすま１０ｇ、リン酸二水素カリウム３ｇ、硫酸マグネシウム七水和物２ｇが含まれ、この培地は自然ｐＨである。
２４～９６時間の発酵中には、濃度２０ｇ／Ｌのフェニルアラニンを連続流加して、７日間、発酵させ、合計１６０ｍＬ流加し、その最終濃度が１ｇ／Ｌとなるようにした。発酵中に、霊芝細胞外多糖の収量を測定した。その結果から明らかなように、霊芝多糖の収量は０．４７１ｇ／Ｌであり、実施例７よりも２８％向上した。
Example 9
Reishi seed culture A bacterial mass having a size of 0.5 cm 2 was collected, inoculated into an Erlenmeyer flask having a liquid charge of 80 mL / 250 mL, and cultured at 150 r · min -1 , 30 ° C. for 7 days.
Fermentation culture of Reishi A 3L fermentation medium was added to a 5L fermentation tank and sterilized at 115 ° C. for 20 minutes. Reishi mycelium having a wet weight of 10 g was inoculated and cultured at 150 r · min -1 with an aeration rate of 1.5 L · min -1 at 30 ° C. for 7 days.
The composition of the seed medium and the fermentation medium is as follows. Each liter contains 20 g of glucose, 10 g of corn flour, 10 g of bran, 3 g of potassium dihydrogen phosphate and 2 g of magnesium sulfate heptahydrate, and this medium has a natural pH.
During the 24-96 hours fermentation, phenylalanine at a concentration of 20 g / L was continuously fed and fermented for 7 days, and a total of 160 mL was fed so that the final concentration was 1 g / L. During fermentation, the yield of Reishi extracellular polysaccharide was measured. As is clear from the results, the yield of Reishi polysaccharide was 0.471 g / L, which was 28% higher than that of Example 7.
実施例１０
アギタケのシード培養
サイズ０．５ｃｍ２の菌塊を採取し、シード培地の液体仕込み量が８０ｍＬ／２５０ｍＬの三角フラスコに接種して、１５０ｒ・ｍｉｎ－１、２８℃で７日間、培養した。
アギタケの発酵培養
５００ｍＬの三角フラスコに発酵培地１５０ｍＬを加え、１１５℃で２０分殺菌した。発酵培地に湿重量０．５ｇの菌糸体を接種して、１５０ｒ・ｍｉｎ－１、２８℃で７日間、培養した。
前記シード培地と発酵培地１の配合は以下のとおりである。１Ｌあたり、グルコース２０ｇ、トリプトン５ｇ、無アミノ基酵母（ＹＮＢ）５ｇ、リン酸二水素カリウム４．５ｇ、硫酸マグネシウム七水和物２ｇが含まれ、この培地は自然ｐＨである。
発酵中に、アギタケ細胞外多糖の収量を測定し、その結果から明らかなように、アギタケ細胞外多糖の収量は０．２０１ｇ／Ｌ発酵上澄み液であった。
Example 10
Seed culture of Agitake A bacterial mass having a size of 0.5 cm 2 was collected, inoculated into an Erlenmeyer flask having a liquid charge of 80 mL / 250 mL, and cultured at 150 r · min -1 at 28 ° C. for 7 days.
Fermentation culture of Agitake 150 mL of fermentation medium was added to a 500 mL Erlenmeyer flask and sterilized at 115 ° C. for 20 minutes. The fermentation medium was inoculated with mycelium having a wet weight of 0.5 g and cultured at 150 r · min -1 , 28 ° C. for 7 days.
The composition of the seed medium and the fermentation medium 1 is as follows. Each L contains 20 g of glucose, 5 g of tryptone, 5 g of amino-free yeast (YNB), 4.5 g of potassium dihydrogen phosphate, and 2 g of magnesium sulfate heptahydrate, and this medium has a natural pH.
During the fermentation, the yield of Agitake extracellular polysaccharide was measured, and as is clear from the results, the yield of Agitake extracellular polysaccharide was 0.201 g / L fermentation supernatant.
実施例１１
０ｈに発酵培地へフェニルアラニンをその最終濃度が１ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１０と同様であった。発酵中に、アギタケ細胞外多糖の収量を測定し、その結果から明らかなように、アギタケ細胞外多糖の収量は０．２６４ｇ／Ｌ発酵上澄み液であり、実施例１０よりも３１％向上した。
Example 11
The medium and culture method were the same as in Example 10 except that phenylalanine was added to the fermentation medium at 0 h so that the final concentration was 1 g / L. During the fermentation, the yield of Agitake extracellular polysaccharide was measured, and as is clear from the results, the yield of Agitake extracellular polysaccharide was 0.264 g / L fermentation supernatant, which was 31% higher than that of Example 10.
実施例１２
実験菌株にはアラゲカワラタケが使用される以外、培地及び培養方法は実施例１０と同様であった。発酵中に、アラゲカワラタケ細胞外多糖の収量を測定し、その結果から明らかなように、アラゲカワラタケ細胞外多糖の収量は０．２８０ｇ／Ｌ発酵上澄み液であった。
Example 12
The medium and culture method were the same as in Example 10 except that Aragekawaratake was used as the experimental strain. During fermentation, the yield of Aragekawaratake extracellular polysaccharide was measured, and as is clear from the results, the yield of Aragekawaratake extracellular polysaccharide was 0.280 g / L fermentation supernatant.
実施例１３
実験菌株にはアラゲカワラタケが使用され、且つ、０ｈに発酵培地へフェニルアラニンをその最終濃度が２ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１０と同様であった。発酵中に、アラゲカワラタケ細胞外多糖の収量を測定し、その結果から明らかなように、アラゲカワラタケ細胞外多糖の収量は０．３２２ｇ／Ｌ発酵上澄み液であり、実施例１２よりも１５％向上した。
Example 13
The medium and culture method were the same as in Example 10 except that Aragekawaratake was used as the experimental strain and phenylalanine was added to the fermentation medium at 0 h so that the final concentration was 2 g / L. During fermentation, the yield of Aragekawaratake extracellular polysaccharide was measured, and as is clear from the results, the yield of Aragekawaratake extracellular polysaccharide was 0.322 g / L fermentation supernatant, which was 15% higher than that of Example 12. did.
実施例１４
実験菌株にはスエヒロタケが使用される以外、培地及び培養方法は実施例１０と同様であった。発酵中に、スエヒロタケ細胞外多糖の収量を測定し、その結果から明らかなように、スエヒロタケ細胞外多糖の収量は０．４５２ｇ／Ｌ発酵上澄み液であった。
Example 14
The medium and culture method were the same as in Example 10 except that Schizophyllum communis was used for the experimental strain. During fermentation, the yield of exopolysaccharide of Schizophyllum communis was measured, and as is clear from the results, the yield of exopolysaccharide of Schizophyllum communis was 0.452 g / L fermentation supernatant.
実施例１５
実験菌株にはスエヒロタケが使用され、且つ、０ｈに発酵培地へフェニルアラニンをその最終濃度が３ｇ／Ｌとなるように添加する以外、培地及び培養方法は実施例１０と同様であった。発酵中に、スエヒロタケ細胞外多糖の収量を測定し、その結果から明らかなように、スエヒロタケ細胞外多糖の収量は０．５６３ｇ／Ｌ発酵上澄み液であり、実施例１４よりも２５％向上した。
Example 15
The medium and culture method were the same as in Example 10 except that Schizophyllum communis was used as the experimental strain and phenylalanine was added to the fermentation medium at 0 h so that the final concentration was 3 g / L. The yield of exopolysaccharide of Schizophyllum communis was measured during fermentation, and as is clear from the results, the yield of exopolysaccharide of Schizophyllum communis was 0.563 g / L fermentation supernatant, which was 25% higher than that of Example 14.
実施例１６
実施例１～１５と同様な方策によって、キクラゲ、シイタケ、チョレイマイタケ、シロキクラゲ、マツホド、カワラタケ、ヤマブシタケ、冬虫夏草などの食用・薬用真菌の発酵中にフェニルアラニンをそれぞれ添加し、その結果から明らかなように、細胞外多糖の収量は、異なる程度の向上を有する。
比較例１
発酵液にアミノ酸を最終濃度が１ｇ／Ｌとなるようにそれぞれ添加する以外、実施例１と同様にして、細胞外多糖の収量を測定し、結果を下記表1に示した。本比較例では、実施例1と比較して、添加したアミノ酸をトリプトファン、アルギニン、ヒスチジン、リジン、イソロイシン、プロリン、バリンに変えた。
By the same measures as in Examples 1 to 15, phenylalanine was added during the fermentation of edible and medicinal fungi such as fungus, shiitake mushroom, polyporus umbellatus, white fungus, pine root, coriolus versicolor, lion's mane, and winter worm summer grass, respectively, as is clear from the results. In addition, the yield of extracellular polysaccharides has different degrees of improvement.
Comparative Example 1
The yield of extracellular polysaccharide was measured in the same manner as in Example 1 except that amino acids were added to the fermentation broth so that the final concentration was 1 g / L, and the results are shown in Table 1 below. In this comparative example, the added amino acids were changed to tryptophan, arginine, histidine, lysine, isoleucine, proline, and valine as compared with Example 1.
本願に記載の食用・薬用真菌は、今のところ知られている細胞外多糖を産生するための食用・薬用真菌に限定されず、既知の食用・薬用真菌に類似性のある既知又は未知の属種の真菌は、いずれも本願の方法に適用でき、細胞外多糖の収量向上には類似した効果を達成することができる。 The edible / medicinal fungi described in the present application are not limited to the edible / medicinal fungi for producing extracellular polysaccharides known so far, and the known or unknown genus having similarities to the known edible / medicinal fungi. All of the fungi of the species can be applied to the methods of the present application and can achieve similar effects in improving the yield of extracellular polysaccharides.
以上、本発明を好適な実施例により開示したが、これらは本発明を限定するものではなく、当業者であれば、本発明の趣旨及びその範囲から逸脱することなく、様々な変更や修飾を行うことができるため、本発明の特許範囲は、特許請求の範囲によって定められるものとする。 Although the present invention has been disclosed by preferred embodiments as described above, these are not limited to the present invention, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the present invention. The scope of the invention of the present invention shall be determined by the scope of claims.
Claims (8)
前記真菌が、霊芝（Ｇａｎｏｄｅｒｍａ ｌｕｃｉｄｕｍ）、アギタケ（Ｐｌｅｕｒｏｔｕｓ ｆｅｒｕｌａｅ）、アラゲカワラタケ（Ｃｏｒｉｏｌｕｓ ｈｉrｓｕｔｕｓ）又はスエヒロタケ（Ｓｃｈｉｚｏｐｈｙｌｌｕｍ ｃｏｍｍｕｎｅ）を含むことを特徴とする真菌細胞外多糖の収量を高める方法。 Adding phenylalanine early in or during fungal fermentation , and
A method for increasing fungal exopolysaccharide, characterized in that the fungus comprises Ganoderma lucidum, Pleurotus ferulae, Coriolus hirsutus or Schizophyllum commune .
前記真菌が、霊芝（Ｇａｎｏｄｅｒｍａ ｌｕｃｉｄｕｍ）、アギタケ（Ｐｌｅｕｒｏｔｕｓ ｆｅｒｕｌａｅ）、アラゲカワラタケ（Ｃｏｒｉｏｌｕｓ ｈｉrｓｕｔｕｓ）又はスエヒロタケ（Ｓｃｈｉｚｏｐｈｙｌｌｕｍ ｃｏｍｍｕｎｅ）を含むことを特徴とする食用・薬用真菌の細胞外多糖の製造方法。 Fermentation is performed by inoculating the fungus into the fermentation medium, and phenylalanine is added to the fermentation medium in 0 to 96 hours of fermentation, and
An edible / medicinal fungus for producing an edible / medicinal fungus, wherein the fungus contains Ganoderma lucidum, Pleurotus ferulae, Coriolus hirsutus, or Schizophyllum commune .
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