JP5800211B1 - Artificial culture method of fruit body - Google Patents
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- JP5800211B1 JP5800211B1 JP2015026299A JP2015026299A JP5800211B1 JP 5800211 B1 JP5800211 B1 JP 5800211B1 JP 2015026299 A JP2015026299 A JP 2015026299A JP 2015026299 A JP2015026299 A JP 2015026299A JP 5800211 B1 JP5800211 B1 JP 5800211B1
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- 235000013399 edible fruits Nutrition 0.000 title claims abstract description 46
- 238000012136 culture method Methods 0.000 title claims description 9
- 239000001963 growth medium Substances 0.000 claims abstract description 30
- 238000000034 method Methods 0.000 claims abstract description 22
- 239000002609 medium Substances 0.000 claims description 19
- 235000007164 Oryza sativa Nutrition 0.000 claims description 8
- 235000009566 rice Nutrition 0.000 claims description 8
- 230000001954 sterilising effect Effects 0.000 claims description 7
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims description 5
- 241000190633 Cordyceps Species 0.000 claims description 4
- 235000019764 Soybean Meal Nutrition 0.000 claims description 4
- 244000144972 livestock Species 0.000 claims description 4
- 239000004455 soybean meal Substances 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 235000001674 Agaricus brunnescens Nutrition 0.000 claims description 3
- 239000004480 active ingredient Substances 0.000 claims description 3
- 239000013589 supplement Substances 0.000 claims description 3
- 238000009423 ventilation Methods 0.000 claims description 3
- 241000894006 Bacteria Species 0.000 claims description 2
- 241000382353 Pupa Species 0.000 claims description 2
- 241001264174 Cordyceps militaris Species 0.000 claims 1
- 240000007594 Oryza sativa Species 0.000 claims 1
- 230000008569 process Effects 0.000 abstract description 11
- 235000013305 food Nutrition 0.000 abstract description 8
- 239000006227 byproduct Substances 0.000 abstract description 5
- 235000015701 Artemisia arbuscula Nutrition 0.000 abstract description 4
- 235000002657 Artemisia tridentata Nutrition 0.000 abstract description 4
- 235000003261 Artemisia vulgaris Nutrition 0.000 abstract description 4
- 240000006891 Artemisia vulgaris Species 0.000 abstract description 4
- 238000012258 culturing Methods 0.000 abstract description 2
- 239000004464 cereal grain Substances 0.000 abstract 1
- 239000002054 inoculum Substances 0.000 description 12
- 235000013339 cereals Nutrition 0.000 description 10
- 238000002360 preparation method Methods 0.000 description 9
- 230000000052 comparative effect Effects 0.000 description 8
- 241000209094 Oryza Species 0.000 description 7
- 239000000203 mixture Substances 0.000 description 6
- 239000000843 powder Substances 0.000 description 5
- 235000017166 Bambusa arundinacea Nutrition 0.000 description 3
- 235000017491 Bambusa tulda Nutrition 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 244000082204 Phyllostachys viridis Species 0.000 description 3
- 235000015334 Phyllostachys viridis Nutrition 0.000 description 3
- 244000061456 Solanum tuberosum Species 0.000 description 3
- 235000002595 Solanum tuberosum Nutrition 0.000 description 3
- 239000011425 bamboo Substances 0.000 description 3
- 230000000975 bioactive effect Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 230000035784 germination Effects 0.000 description 3
- 239000011521 glass Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 238000010438 heat treatment Methods 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 229920001817 Agar Polymers 0.000 description 2
- KQLDDLUWUFBQHP-UHFFFAOYSA-N Cordycepin Natural products C1=NC=2C(N)=NC=NC=2N1C1OCC(CO)C1O KQLDDLUWUFBQHP-UHFFFAOYSA-N 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- 206010017533 Fungal infection Diseases 0.000 description 2
- 241000233866 Fungi Species 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 239000013065 commercial product Substances 0.000 description 2
- OFEZSBMBBKLLBJ-BAJZRUMYSA-N cordycepin Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)C[C@H]1O OFEZSBMBBKLLBJ-BAJZRUMYSA-N 0.000 description 2
- OFEZSBMBBKLLBJ-UHFFFAOYSA-N cordycepine Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)CC1O OFEZSBMBBKLLBJ-UHFFFAOYSA-N 0.000 description 2
- 239000002537 cosmetic Substances 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000003203 everyday effect Effects 0.000 description 2
- 208000024386 fungal infectious disease Diseases 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 235000013402 health food Nutrition 0.000 description 2
- 238000010979 pH adjustment Methods 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 description 1
- 235000001270 Allium sibiricum Nutrition 0.000 description 1
- 229920002498 Beta-glucan Polymers 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
- 241000222350 Pleurotus Species 0.000 description 1
- 235000013999 Prunus japonica Nutrition 0.000 description 1
- 241000392950 Prunus japonica Species 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 239000003674 animal food additive Substances 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 235000013527 bean curd Nutrition 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
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- 239000010794 food waste Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000003308 immunostimulating effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 239000013028 medium composition Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 150000002989 phenols Chemical class 0.000 description 1
- 239000000047 product Substances 0.000 description 1
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- 239000010902 straw Substances 0.000 description 1
- 230000000304 vasodilatating effect Effects 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Landscapes
- Mushroom Cultivation (AREA)
- Fodder In General (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
【課題】食品工業副産物を利用したサナギタケ子実体の培養培地、さらにサナギタケ子実体の人工培養方法を提供する。【解決手段】本発明に係るサナギタケ子実体の培養培地は、おからを主成分とし、さらに少なくとも、穀物の糠、油粕を含むことを特徴とするサナギタケ子実体の培養培地とした。また、本発明に係るサナギタケ子実体の人工培養方法は、食品工業由来のおからを主成分とする滅菌した培養培地に、サナギタケの菌糸培養液を接種し、それぞれ異なる培養環境である、菌糸体培養工程、芽出し工程、子実体育成工程を経てサナギタケの子実体を培養する方法である。【選択図】図１The present invention provides a culture medium for the fruit body of Sanagitake using a food industry by-product, and a method for artificial culture of the fruit body of Sanagitake. The culture medium for the fruit body of Sanagitake according to the present invention is a culture medium for the fruit body of Sanagitake, characterized in that it contains okara as a main component and further contains at least cereal grains and oil cake. In addition, the method for artificially cultivating fruit bodies of the present invention comprises inoculating a sterilized culture medium mainly composed of okara derived from the food industry with a mycelium culture medium of sanagitake, and mycelia that are different culture environments. This is a method for cultivating the fruit body of a sagebrush through a culturing process, a sprouting process, and a fruiting body growing process. [Selection] Figure 1
Description
本発明は、食品工業副産物を利用したサナギタケ子実体の培養培地、さらに野生のサナギタケ子実体と同様の生理活性成分を含有させつつ、容易、低廉かつ迅速に培養し、健康食品、医薬品、化粧品等に用いることができるサナギタケ子実体の人工培養方法に関する。 The present invention is a culture medium for the fruit body of Sanagitake using the food industry by-product, and further, easily, inexpensively and rapidly cultured while containing the same physiologically active components as the fruit body of wild Sanagitake, health food, pharmaceuticals, cosmetics, etc. The present invention relates to a method for artificially cultivating fruit bodies of the sagebrush that can be used in the field.
サナギタケ（蛹茸 学名：Ｃｏｒｄｙｃｅｐｓ ｍｉｌｉｔａｒｉｓ（Ｖｕｉｌｌ．）Ｆｒ．）は、冬虫夏草属に属する菌類の一種であり、野生ではチョウ目の幼虫や蛹に菌が感染し、体内に菌糸体が蔓延し、その菌糸体から子実体（キノコ部）が発生する。サナギタケ子実体はオレンジ色の棍棒状である。 Sanatake mushroom (Scientific name: Cordyceps militaryis (Vuill.) Fr.) is a kind of fungus belonging to the genus Cordyceps. Fruit bodies (mushrooms) are generated from the mycelium. Sanagitake fruiting body has an orange club shape.
サナギタケ子実体には、抗腫瘍性を示すコルジセピンをはじめ、血管拡張作用を有するとされるＤ−マンニトール、免疫賦活作用があるとされるβ−グルカン、活性酸素を消去するＳＯＤなどの生理活性成分が豊富に含まれていることが知られている。 The fruit body of Sanagitake includes physiologically active components such as cordycepin showing antitumor properties, D-mannitol, which is said to have a vasodilatory effect, β-glucan, which is said to have an immunostimulatory effect, and SOD which eliminates active oxygen Is known to be abundant.
そのため、昨今、絶滅状態にあった冬虫夏草（Ｃｏｒｄｙｃｅｐｓ ｓｉｎｅｎｓｉｓ（Ｂｅｒｋｅｌｅｙ） Ｓａｃｃａｒｄｏ）の代用品として世界中に注目されている。 Therefore, it has been attracting attention all over the world as a substitute for Cordyceps sinensis (Berkeley) Sacccardo, which has been extinct recently.
サナギタケ子実体を人工的に形成させる研究は、米国などの各国で成功例が報告されており、特に中国では人工的に形成したサナギタケ子実体を漢方薬剤として使用する研究が活発になされ、現在では実用段階にある。また、韓国内でも人工サナギタケ子実体を形成する研究が近年活発になされており、既に商業的目的で利用するための多様な研究が進行中である。 There have been reports of successful examples of the formation of fruit bodies of sanagitake in various countries such as the United States, especially in China, where research on the use of fruit bodies of sanagitake artificially formed as a traditional Chinese medicine has been active. It is in a practical stage. In Korea, research on the formation of artificial sanagitake fruit bodies has been actively carried out in recent years, and various researches for use for commercial purposes are already underway.
しかしながら、特許文献１、２等で開示されているように、人工的なサナギタケ子実体の培養培地の構成が複雑で、コストが高く、これまでのサナギタケ子実体の人工培養方法は普及していない。 However, as disclosed in Patent Documents 1 and 2 and the like, the structure of the artificial culture medium for the fruit body of Sanagitake is complex and high in cost, and the conventional artificial culture method for the fruit body of Sanagitake is not widespread. .
そこで、本発明は、食品工業副産物を利用したサナギタケ子実体の培養培地、野生のサナギタケ子実体と同様の生理活性成分を含有させつつ、容易、低廉かつ迅速に培養し、健康食品、医薬品、化粧品等に用いることができるサナギタケ子実体の人工培養方法を提供することを目的とする。 Accordingly, the present invention provides a culture medium for the fruit body of Sanagitake using the food industry by-product, a physiologically active ingredient similar to that of the wild fruit tree, and easily, inexpensively and rapidly cultured, It is an object of the present invention to provide a method for artificially cultivating the fruit body of Pleurotus that can be used for the like.
上述の課題を解決するため本発明の構成は、
（１）
おからを主成分とし、さらに少なくとも、穀物の糠、油粕を含むことを特徴とするサナギタケ子実体の培養培地。
（２）
おからを主成分とし、さらに少なくとも、穀物の茎の粉末、油粕を含むことを特徴とするサナギタケ子実体の培養培地。
（３）
おから５０〜９０重量部と、
穀物の糠又は穀物の茎の粉末４０〜９重量部と、
油粕１０〜１重量部からなることを特徴とする
サナギタケ子実体の培養培地。
（４）
（１）〜（３）の何れかに記載のサナギタケ子実体の培養培地を、滅菌した後、サナギタケの菌糸培養液を前記滅菌培地に接種し、
培養環境条件を、空中湿度６０％以上、温度２０−２５℃及び暗条件として、サナギタケ菌糸体を培養し、
前記サナギタケ菌糸体が前記滅菌培地の略全体に蔓延した後、
培養環境条件を、空中湿度８０％以上、１５℃以上２０℃より低くい温度及び８０−２５０Ｌｕｘの光強度の間欠若しくは連続的な光照射に切り替え、サナギタケ子実体の芽を発育させ、
前記サナギタケ子実体の芽が出た後、
培養環境条件を、空中湿度８０％以上、温度２０−２５℃、間欠的な換気、及び１５０Ｌｕｘ以上の光強度の間欠若しくは連続的な照射に切り替え、サナギタケ子実体を育成することを特徴とする
サナギタケ子実体の人工培養方法。
（５）
前記サナギタケが、Ｃｏｒｄｙｃｅｐｓ属に属する菌であることを特徴とする（４）に記載のサナギタケの人工培養方法。
（６）
（４）又は（５）に記載のサナギタケ子実体の人工培養方法で培養したサナギタケ子実体及び／又はそれから抽出した生理活性成分を含むことを特徴とするサプリメント。
（７）
（４）又は（５）に記載のサナギタケ子実体の人工培養方法で発生したことを特徴とする菌糸体を含む菌床。
（８）
（７）に記載の菌糸体を含む菌床を用いて作成されたことを特徴とする家畜飼料。
とした。
In order to solve the above problems, the configuration of the present invention is as follows.
(1)
A culture medium for fruit bodies of chives, comprising okara as a main component and further containing at least cereal and oil cakes.
(2)
A culture medium for the fruit body of Prunus japonica, comprising okara as a main component, and further comprising at least cereal stem powder and oil cake.
(3)
50 to 90 parts by weight of okara,
40-9 parts by weight of cereal bran or cereal stalk powder;
A culture medium for the fruit body of the sagebrush, comprising 10 to 1 part by weight of oil cake.
(4)
(1) After sterilizing the culture medium of the fruit body of Sanagitake according to any of (3), inoculating the mycelium culture medium of Sanagitake to the sterile medium,
Cultivating Sanagitake mycelium with the culture environment conditions of air humidity 60% or higher, temperature 20-25 ° C. and dark conditions,
After the Sanagitake mycelium has spread throughout substantially the entire sterile medium,
The culture environment condition is switched to an air humidity of 80% or higher, a temperature not lower than 15 ° C. and lower than 20 ° C., and intermittent or continuous light irradiation with a light intensity of 80-250 Lux.
After the shoots of the pupa
The culture environment condition is changed to an air humidity of 80% or higher, a temperature of 20-25 ° C., intermittent ventilation, and intermittent or continuous irradiation with light intensity of 150 Lux or higher, and the fruit body of the water fly is cultivated. Artificial culture method for fruiting bodies.
(5)
The method for artificial culture of sanagitake according to (4), wherein the sanagitake is a bacterium belonging to the genus Cordyceps .
(6)
A supplement comprising the fruit body of Sanagitake cultivated by the artificial culture method for fruit bodies of Sanagitake according to (4) or (5) and / or a physiologically active ingredient extracted therefrom.
(7)
A fungus bed comprising a mycelium, which is generated by the method for artificially cultivating a fruit body of Sanagitake according to (4) or (5).
(8)
A livestock feed produced using a mycelium containing the mycelium according to (7).
It was.
本発明であるはサナギタケ子実体の人工培養方法では、食品工業由来のおから、さらには、少なくとも穀物の糠又は穀物の茎の粉末、及び油粕の内から選ばれる１種または２種以上を含む培地を滅菌し、サナギタケの菌糸培養液を接種し、菌糸体培養工程、芽出し工程、育成工程を経てサナギタケ子実体を培養するもので、既存の人工培養方法（例えば：特許文献１、２）よりも容易、低廉かつ迅速にサナギタケ子実体を培養することができる。 In the method for artificial culture of the fruit tree of the present invention, it includes okara derived from the food industry, and at least one or more selected from among cereal straw or cereal powder and oil cake. Sterilize the medium, inoculate the mycelium culture solution of sanagitake, and cultivate the fruit body of sanagitake through the mycelium culture process, the sprouting process, and the growing process. From the existing artificial culture methods (for example: Patent Documents 1 and 2) In addition, it is possible to cultivate the fruit body of Sanagitake easily, inexpensively and quickly.
本発明によれば、野生のサナギタケと同様の形態の子実体を培養することができる。また、野生のサナギタケ子実体と同様の生理活性成分量を含有するサナギタケ子実体を効率よく安価に得ることができる。この人工培養サナギタケ子実体をそのまま粉砕し加工したり、必要に応じて、生理活性成分を抽出したりして、医薬品、サプリメントなどの健康食品、化粧品などに利用することができる。 According to the present invention, a fruiting body having the same form as that of wild bamboo shoots can be cultured. In addition, it is possible to efficiently and inexpensively obtain the fruit body of Sanagitake, which contains the same amount of physiologically active components as the fruit body of wild Sanagitake. The artificially cultured Sanagitake fruiting body can be crushed and processed as it is, or a physiologically active component can be extracted as necessary, and used for health foods such as pharmaceuticals and supplements, cosmetics, and the like.
さらには、菌糸体（菌糸塊）にも、多糖類、フェノール類、ＳＯＤ様物質などのサナギタケの特有の生理活性成分、その他機能生物質が含有するため、培養残渣である培地菌糸体を含み、家畜を対象とした飼料の添加物に応用することもできる。 Furthermore, the mycelium (mycelium mass) also contains a medium mycelium that is a culture residue, because it contains polysaccharides, phenols, bioactive components peculiar to bamboo shoots such as SOD-like substances, and other functional biomaterials. It can also be applied to feed additives for livestock.
つまり、本発明は、食品工業副産物を利用してサナギタケ子実体を培養することで食品廃棄物を減少させることができるとともに、培養済みの菌糸体を含む培地は家畜飼料にすることで、培地を廃棄物とすることのない、環境にやさしいサナギタケ子実体の人工培養方法を提供することができる。 That is, according to the present invention, food waste can be reduced by culturing Sanagitake fruiting bodies using food industry by-products, and the medium containing the cultured mycelium is used as a livestock feed. It is possible to provide an environment-friendly artificial culture method for the fruit body of Sanagitake that does not become waste.
以下、本発明を実施するための形態を実施例及び図面に基づいて具体的に説明する。なお、本発明はそれら実施形態に限定されるものではない。 DESCRIPTION OF EMBODIMENTS Hereinafter, embodiments for carrying out the present invention will be specifically described based on examples and drawings. Note that the present invention is not limited to these embodiments.
実施の一例であるサナギタケ子実体の人工培養方法は、以下の手順で行った。 An example of the method for artificially cultivating the fruit body of the sagebrush was as follows.
１）［試管培地の作製］
・試管培地組成
グルコース ２０ｇ
寒天 ２０ｇ
ペプトン １０ｇ
ポテトエキス ４ｇ
ＫＨ２ＰＯ４ ３ｇ
ＭｇＳＯ４・７Ｈ２Ｏ １．５ｇ
蒸留水 １０００ｍｌ
1) [Preparation of tube culture medium]
・ Test tube medium composition Glucose 20g
Agar 20g
Peptone 10g
Potato extract 4g
KH 2 PO 4 3g
MgSO 4 · 7H 2 O 1.5g
1000ml distilled water
グルコース、寒天、ペプトン、ポテトエキス、ＫＨ２ＰＯ４、ＭｇＳＯ４・７Ｈ２Ｏは市販品を用いた（以下同じ）。 Glucose, agar, peptone, potato extract, KH 2 PO 4 , MgSO 4 .7H 2 O were commercially available products (the same applies hereinafter).
・試管培地調整
前記試管培地の組成物を攪拌した（ｐＨ調整なし）。これを１２１℃に設定した殺菌釜で２０分間加熱殺菌（オートクレーブ）し、前記組成を１５ｍｌほど試験管に流し込み、水平から約１０°に傾けたまま冷却ゲル化させ、固形の試管培地を得た。なお、培地の殺菌は、オートクレーブ以外の方法であってもよい（以下同様）。
-Preparation of test tube medium The composition of the test tube medium was stirred (no pH adjustment). This was sterilized by heating (autoclave) for 20 minutes in a sterilization pot set at 121 ° C., about 15 ml of the composition was poured into a test tube, and cooled and gelled while tilting at about 10 ° from the horizontal to obtain a solid test tube medium. . The medium may be sterilized by a method other than autoclaving (the same applies hereinafter).
２）［試管種菌（菌母）の作製］
市販の種菌（スラント培地培養の菌糸状種菌）を、前記１）［試管培地の作製］で作製した試管培地（ゲル）に移植して菌糸体を培養し、試管種菌（菌母）を完成させる。
2) [Preparation of test tube inoculum (mycosis mother)]
A commercially available inoculum (mycelium inoculum in slant medium culture) is transplanted to the test medium (gel) prepared in 1) [Preparation of test medium], and the mycelium is cultured to complete the test inoculum (mycosis mother). .
３）［培養種菌の作製］
・種菌培養液体培地（以下「菌糸培養液」という）組成
グルコース ２０ｇ
ペプトン １０ｇ
ポテトエキス ４ｇ
ＫＨ２ＰＯ４ ３ｇ
ＭｇＳＯ４・７Ｈ２Ｏ １．５ｇ
蒸留水 １０００ｍｌ
3) [Preparation of cultured inoculum]
Inoculum culture liquid medium (hereinafter referred to as “mycelium culture solution”) composition glucose 20 g
Peptone 10g
Potato extract 4g
KH 2 PO 4 3g
MgSO 4 · 7H 2 O 1.5g
1000ml distilled water
・菌糸培養液の調整
前記菌糸培養液組成を攪拌した（ｐＨ調整なし）。これを１２１℃に設定した殺菌釜で２０分間加熱殺菌（オートクレーブ）し、クリーンベンチに移し替えて室温まで冷却し、菌糸培養液を作製した。
-Preparation of mycelium culture solution The composition of the mycelium culture solution was stirred (no pH adjustment). This was sterilized by heating (autoclave) for 20 minutes in a sterilization pot set at 121 ° C., transferred to a clean bench, and cooled to room temperature to prepare a mycelia culture solution.
この菌糸培養液に、前記２）［試管種菌（菌母）の作製］で作成し、菌母を接種して菌糸体が十分に回って白変した試管培地を移し入れる。その分量は菌糸培養液１０００ｍｌに対して試管種菌１本（１５ｍｌ／ゲル）の割合とした。その後、温度を２３℃に設定して電磁攪拌培養した。約１週間が経過する頃、菌糸培養液中に球状の菌糸体（培養種菌）の広がりが確認できた。 To this mycelium culture solution, the tube medium prepared in 2) [Preparation of test tube inoculum (mycelium)], inoculated with mycelia, and the mycelium turned sufficiently white is transferred. The amount was set to a ratio of one tube inoculum (15 ml / gel) to 1000 ml of the mycelium culture solution. Thereafter, the temperature was set to 23 ° C. and the magnetic stirring culture was performed. When about 1 week passed, the spread of spherical mycelium (cultured inoculum) was confirmed in the mycelium culture solution.
４）［サナギタケ子実体の培養培地の作製］
・培養培地組成
おから ９０５ｇ（約９０％）
米糠 ８６ｇ（約９％）
大豆かす ９ｇ（約１％）
合計１０００ｇ（１００％）
4) [Preparation of culture medium of bamboo fruit body]
・ Culture medium composition Okara 905g (about 90%)
86g of rice bran (about 9%)
Soybeans 9g (about 1%)
Total 1000g (100%)
おからは豆腐店から入手した。米糠は穀物の糠の例として、精米所から若しくは市販品を入手した。大豆かすは油粕の例として、市販品を入手した。おからと米糠は乾燥せず、そのまま使用した。大豆かすは乾燥して、粉砕して使用した。米糠の他、他の穀物糠も使用でき、さらに穀物の茎の粉末であってもよい。穀物の茎の粉末であれば、米糠などのような他の食品等へ利用が全くない未利用バイオマスであるので、極めて安価に入手でき、培養コストの抑制に繋がり好ましい。また大豆かすを他の油粕に置換してもよい。 Okara was obtained from a tofu store. Rice bran was obtained from rice mills or as a commercial product as an example of grain koji. Soybean meal was obtained as a commercial product as an example of oil cake. Okara and rice bran were not dried and used as they were. The soybean meal was dried and ground before use. In addition to rice bran, other grain bran can be used, and a grain stem powder may be used. Grain stalk powder is preferable because it is an unused biomass that is not used for other foods such as rice bran, and can be obtained at a very low cost, leading to a reduction in culture costs. In addition, soybean meal may be replaced with another oil cake.
・培養培地の調整
前記の培養培地組成成分を混合し、ガラス瓶４（容量：３６８ｍｌ、直径：７１ｍｍ、高さ：１２２ｍｍ）に、混合培養培地を５０ｇ入れ、通気性を有するキャップ５を被着し、１２１℃に設定した殺菌釜で２０分間加熱殺菌した。その後、容器を殺菌釜から取り出してクリーンベンチに運び入れ、前記キャップ５を開けて、紫外線ランプ照射下で、培養培地を自然的に冷却した。
-Preparation of culture medium The above culture medium composition components were mixed, 50 g of mixed culture medium was put into a glass bottle 4 (volume: 368 ml, diameter: 71 mm, height: 122 mm), and a cap 5 having air permeability was attached. And sterilized by heating in a sterilization pot set at 121 ° C. for 20 minutes. Thereafter, the container was taken out of the sterilization pot and carried to a clean bench, the cap 5 was opened, and the culture medium was naturally cooled under irradiation with an ultraviolet lamp.
・種菌接種
培養培地の温度が２５℃を下回ったことを確認してから、前記３）で作製した培養種菌（培地重量比５％−１０％）を培養培地に接種した。
Inoculum Inoculation After confirming that the temperature of the culture medium was below 25 ° C, the culture inoculum (medium weight ratio 5% -10%) prepared in 3) above was inoculated into the culture medium.
５）［培養］
・菌糸体培養工程
培養種菌を接種した培養培地を温度２０−２５℃、湿度６０％以上に設定した培養室で、暗条件下で培養した。暗条件下で培養すると、菌糸体が培養培地全体に蔓延し、茶色の培養培地が乳白色に変化する。
5) [Culture]
-Mycelium culture step The culture medium inoculated with the culture inoculum was cultured under dark conditions in a culture chamber set at a temperature of 20-25 ° C and a humidity of 60% or more. When cultured under dark conditions, mycelium spreads throughout the culture medium and the brown culture medium turns milky white.
・芽出し工程
培地の表面に米粒状の突起（子実体原基）が確認できたら、１５℃以上２０℃より低くい温度で８０−２５０Ｌｕｘの照明を１２時間間欠若しくは連続的に照射して子実体の発芽を促進させ、突起がオレンジ色に変化する発芽を確認した。光源として、ＬＥＤ、蛍光ランプ若しくは自然光などが例示できる。
・ Sprouting process When rice granular protrusions (fruit body primordia) are confirmed on the surface of the culture medium, 80-250 Lux illumination is irradiated intermittently or continuously for 12 hours at a temperature of 15 ° C. or higher and lower than 20 ° C. Germination was confirmed, and germination in which the protrusions changed to orange was confirmed. Examples of the light source include an LED, a fluorescent lamp, and natural light.
・子実体育成工程
発芽した後、空中湿度８０％以上、温度２０−２５℃、毎日３０分間の換気、毎日１２時間の１５０Ｌｕｘ以上の間欠若しくは連続的な照射条件に切り替え、培養を継続した。
-Fruiting body growing process After germination, the culture was continued by switching to an air humidity of 80% or more, a temperature of 20-25 ° C, ventilation for 30 minutes every day, and intermittent or continuous irradiation conditions of 150 lux or more for 12 hours every day.
６）［成熟／収穫（培養終了）］
サナギタケ子実体の成熟は、子実体と子嚢殻の形成が確認された時点とし、その前後に収穫を行った。
6) [Maturation / harvest (culture end)]
The maturity of the fruit body of Sanagitake was at the time when the formation of the fruit body and the ascollis was confirmed, and harvesting was performed before and after that.
収穫したサナギタケ子実体の生理活性物質を測定し、従来の培地（比較例３、２）、野生採取（比較例１）の含有量と比較した結果を図４に示した。実施例では、アデノシンにおいては比較例３の３．５−４倍、コルジセピンにおいては比較例１の約３倍、比較例２、３の約１０倍に及ぶものもあった。 FIG. 4 shows the result of measuring the physiologically active substance of the harvested fruit body of Sanagitake and comparing it with the contents of the conventional medium (Comparative Examples 3 and 2) and wild collection (Comparative Example 1). In Examples, adenosine was 3.5-4 times that of Comparative Example 3, cordycepin was about 3 times that of Comparative Example 1, and about 10 times that of Comparative Examples 2 and 3.
多糖類においては、野生型の半分程度であるものの、比較例２と同等、比較例３の約５倍の含有量であった。 In the polysaccharide, although it was about half of the wild type, it was equivalent to Comparative Example 2 and about 5 times the content of Comparative Example 3.
本発明は、食品工業副産物のおから等の再利用、さらに機能性物質の生産に貢献できる。 INDUSTRIAL APPLICABILITY The present invention can contribute to the reuse of food industry by-products such as okara and the production of functional substances.
１ サナギタケ菌糸体
２ 子実体原基
３ サナギタケ子実体
４ ガラス瓶
５ キャップ
1 Sanagitake mycelium 2 Fruiting body primitive 3 Sanagitake fruiting body 4 Glass bottle 5 Cap
Claims (5)
培養環境条件を、空中湿度６０％以上、温度２０−２５℃及び暗条件として、サナギタケ菌糸体を培養し、
前記サナギタケ菌糸体が前記滅菌培地の略全体に蔓延した後、
培養環境条件を、空中湿度８０％以上、１５℃以上２０℃より低くい温度及び８０−２５０Ｌｕｘの光強度の間欠若しくは連続的な光照射に切り替え、サナギタケ子実体の芽を発育させ、
前記サナギタケ子実体の芽が出た後、
培養環境条件を、空中湿度８０％以上、温度２０−２５℃、間欠的な換気、及び１５０Ｌｕｘ以上の光強度の間欠若しくは連続的な照射に切り替え、サナギタケ子実体を育成することを特徴とする
サナギタケ子実体の人工培養方法。 After sterilizing the culture medium of Sanagitake fruiting body consisting of 905 parts by weight of okara, 86 parts by weight of rice bran and 9 parts by weight of soybean meal, the sterilization medium is inoculated with the mycelium culture medium of Sanagitake,
Cultivating Sanagitake mycelium with the culture environment conditions of air humidity 60% or higher, temperature 20-25 ° C. and dark conditions,
After the Sanagitake mycelium has spread throughout substantially the entire sterile medium,
The culture environment condition is switched to an air humidity of 80% or higher, a temperature not lower than 15 ° C. and lower than 20 ° C., and intermittent or continuous light irradiation with a light intensity of 80-250 Lux.
After the shoots of the pupa
The culture environment condition is changed to an air humidity of 80% or higher, a temperature of 20-25 ° C., intermittent ventilation, and intermittent or continuous irradiation with light intensity of 150 Lux or higher, and the fruit body of the water fly is cultivated. Artificial culture method for fruiting bodies.
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