WO2022088280A1 - 一种蚕蛹虫草的培养方法 - Google Patents
一种蚕蛹虫草的培养方法 Download PDFInfo
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- WO2022088280A1 WO2022088280A1 PCT/CN2020/129034 CN2020129034W WO2022088280A1 WO 2022088280 A1 WO2022088280 A1 WO 2022088280A1 CN 2020129034 W CN2020129034 W CN 2020129034W WO 2022088280 A1 WO2022088280 A1 WO 2022088280A1
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- silkworm
- parts
- cordyceps militaris
- controlled
- culturing
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Links
- 241000255789 Bombyx mori Species 0.000 title claims abstract description 53
- 241001264174 Cordyceps militaris Species 0.000 title claims abstract description 34
- 238000000034 method Methods 0.000 title claims abstract description 22
- 241000382353 Pupa Species 0.000 claims abstract description 10
- 239000007788 liquid Substances 0.000 claims abstract description 7
- 239000001963 growth medium Substances 0.000 claims abstract description 6
- 238000011081 inoculation Methods 0.000 claims abstract description 5
- 238000012258 culturing Methods 0.000 claims description 13
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
- VZCYOOQTPOCHFL-OWOJBTEDSA-N Fumaric acid Chemical compound OC(=O)\C=C\C(O)=O VZCYOOQTPOCHFL-OWOJBTEDSA-N 0.000 claims description 6
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 claims description 6
- 229910052709 silver Inorganic materials 0.000 claims description 6
- 239000004332 silver Substances 0.000 claims description 6
- 229920001817 Agar Polymers 0.000 claims 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 claims description 3
- 239000001888 Peptone Substances 0.000 claims description 3
- 108010080698 Peptones Proteins 0.000 claims description 3
- 240000008042 Zea mays Species 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 235000015278 beef Nutrition 0.000 claims description 3
- YYRMJZQKEFZXMX-UHFFFAOYSA-N calcium;phosphoric acid Chemical compound [Ca+2].OP(O)(O)=O.OP(O)(O)=O YYRMJZQKEFZXMX-UHFFFAOYSA-N 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- JZCCFEFSEZPSOG-UHFFFAOYSA-L copper(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Cu+2].[O-]S([O-])(=O)=O JZCCFEFSEZPSOG-UHFFFAOYSA-L 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000001530 fumaric acid Substances 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 3
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 3
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 3
- 239000002426 superphosphate Substances 0.000 claims description 3
- VZCYOOQTPOCHFL-UHFFFAOYSA-N trans-butenedioic acid Natural products OC(=O)C=CC(O)=O VZCYOOQTPOCHFL-UHFFFAOYSA-N 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 2
- KQLDDLUWUFBQHP-UHFFFAOYSA-N Cordycepin Natural products C1=NC=2C(N)=NC=NC=2N1C1OCC(CO)C1O KQLDDLUWUFBQHP-UHFFFAOYSA-N 0.000 abstract description 4
- 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 abstract description 4
- OFEZSBMBBKLLBJ-UHFFFAOYSA-N cordycepine Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(CO)CC1O OFEZSBMBBKLLBJ-UHFFFAOYSA-N 0.000 abstract description 4
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 abstract description 3
- 239000004472 Lysine Substances 0.000 abstract description 3
- 239000003242 anti bacterial agent Substances 0.000 abstract description 3
- 229940088710 antibiotic agent Drugs 0.000 abstract description 3
- 239000000122 growth hormone Substances 0.000 abstract description 3
- 230000008569 process Effects 0.000 abstract description 3
- 235000019750 Crude protein Nutrition 0.000 abstract 1
- 239000002253 acid Substances 0.000 abstract 1
- 241000190633 Cordyceps Species 0.000 description 5
- 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 3
- 239000002609 medium Substances 0.000 description 3
- FPVKHBSQESCIEP-UHFFFAOYSA-N (8S)-3-(2-deoxy-beta-D-erythro-pentofuranosyl)-3,6,7,8-tetrahydroimidazo[4,5-d][1,3]diazepin-8-ol Natural products C1C(O)C(CO)OC1N1C(NC=NCC2O)=C2N=C1 FPVKHBSQESCIEP-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 229960002340 pentostatin Drugs 0.000 description 2
- FPVKHBSQESCIEP-JQCXWYLXSA-N pentostatin Chemical compound C1[C@H](O)[C@@H](CO)O[C@H]1N1C(N=CNC[C@H]2O)=C2N=C1 FPVKHBSQESCIEP-JQCXWYLXSA-N 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- -1 t Species 0.000 description 2
- 241000233866 Fungi Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241001248610 Ophiocordyceps sinensis Species 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G18/00—Cultivation of mushrooms
- A01G18/20—Culture media, e.g. compost
Definitions
- the invention relates to the technical field of Cordyceps sinensis, in particular to a method for culturing Cordyceps militaris.
- Cordyceps militaris is a combination of insect and fungus artificially cultivated with wild natural Cordyceps militaris strain spores inoculated on silkworm chrysalis.
- Cordyceps militaris contains more cordycepin, cordycepic acid and SOD than natural cordyceps, northern cordyceps and silkworm cordyceps.
- the purpose of this invention is to provide a kind of preparation method of silkworm cordyceps militaris concentrate, it can improve the output and quality of silkworm cordyceps militaris.
- the technical scheme of the present invention is to design a culture method of Cordyceps militaris, comprising the following steps:
- step 2) before the injection, the silkworm pupae are pierced several times with a silver needle.
- the outer diameter of the silver needle is 2-3 mm.
- the liquid strain is obtained by shaking culture of Cordyceps militaris strains inserted into the culture medium.
- the medium is composed of the following components: 22-27 parts of glucose, 2.7-4.3 parts of ethanol, 2.3-3.8 parts of beef peptone, 0.7-1.4 parts of superphosphate, 3.6-5.7 parts of corn Peptide, 0.8-1.3 parts of magnesium sulfate, 1.8-2.6 parts of potassium dihydrogen phosphate, 1.2-1.8 parts of agar, 0.1-0.3 parts of fumaric acid, 0.1-0.2 parts of copper sulfate pentahydrate, 1.5-2.1 parts of yeast extract , 100 parts of distilled water.
- the temperature of the shaking culture is controlled at 18-20°C, and the time is controlled at 15-20 days.
- the temperature of the dark culture is controlled at 17-20°C
- the humidity is controlled at 55%-60%
- the time is controlled at 6-9 days.
- the temperature of the light culture is controlled at 16-19°C
- the humidity is controlled at 60%-75%
- the time is controlled at 20-25 days.
- the invention pierces the silkworm chrysalis for many times, which can shorten the grass-emergence time of the silkworm cordyceps militaris, and at the same time can increase the grass-emergence amount, and can also improve the effectiveness of cordycepin, cordycepic acid, cordyceps polysaccharide, SOD enzyme, pentostatin and the like in the silkworm cordyceps militaris. content of ingredients.
- the method for culturing silkworm Cordyceps militaris of the present invention uses fresh living silkworm pupa as a culture medium, does not add any growth hormones and antibiotics, and has no pollution in the whole process.
- %, lysine is not less than 2.478%, significantly higher than other Cordyceps militaris, two-way regulation, yin and yang complementarity, more comprehensive nutrition, and more significant efficacy.
- the invention provides a method for culturing Cordyceps militaris, comprising the following steps:
- the outer diameter of the silver needle is 2 to 3 mm;
- the liquid strain is obtained by the shaking culture of the Cordyceps militaris strain of the access medium;
- the medium is composed of the following components: 22-27 parts of glucose, 2.7-4.3 parts of ethanol, 2.3-3.8 parts of beef peptone, 0.7-1.4 parts of superphosphate, 3.6-5.7 parts of corn peptides, 0.8-1.3 parts of parts of magnesium sulfate, 1.8 to 2.6 parts of potassium dihydrogen phosphate, 1.2 to 1.8 parts of agar, 0.1 to 0.3 parts of fumaric acid, 0.1 to 0.2 parts of copper sulfate pentahydrate, 1.5 to 2.1 parts of yeast extract, and 100 parts of distilled water;
- the temperature of the shaking culture is controlled at 18-20°C, and the time is controlled at 15-20 days;
- the temperature of dark culture is controlled at 17-20°C, the humidity is controlled at 55%-60%, and the time is controlled at 6-9 days;
- Light culture light culture the silkworm chrysalis with white hyphae on the surface until the hyphae turn from white to orange;
- the temperature of light culture was controlled at 16-19°C, the humidity was controlled at 60%-75%, and the time was controlled at 20-25 days.
- the invention pierces the silkworm chrysalis for many times, which can shorten the grass-emergence time of the silkworm cordyceps militaris, and at the same time can increase the grass-emergence amount, and can also improve the effectiveness of cordycepin, cordycepic acid, cordyceps polysaccharide, SOD enzyme, pentostatin and the like in the silkworm cordyceps militaris. content of ingredients.
- the method for culturing silkworm Cordyceps militaris of the present invention uses fresh living silkworm pupa as a culture medium, does not add any growth hormones and antibiotics, and has no pollution in the whole process.
- %, lysine is not less than 2.478%, significantly higher than other Cordyceps militaris, two-way regulation, yin and yang complementarity, more comprehensive nutrition, and more significant efficacy.
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Mycology (AREA)
- Environmental Sciences (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Mushroom Cultivation (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
一种蚕蛹虫草的培养方法，包括如下步骤：1)挑选蚕蛹：挑选鲜活的蚕蛹；2)接种：用注射器将液体菌种注射至蚕蛹体内；3)暗培养：将接种后的蚕蛹放入恒温培养箱中避光培养，至蚕蛹表面长出白色菌丝；4)光培养：对表面长出白色菌丝的蚕蛹进行光照培养，至菌丝由白色转为橘黄色。该方法能提高蚕蛹虫草的产量和质量，且以新鲜活体桑蚕蛹为培养基，不添加任何生长激素和抗生素，全程无污染，培养所得的蚕蛹虫草中含有虫草酸不低于4.77%，粗蛋白不低于63.6%，赖氨酸不低于2.478%。
Description
本发明涉及虫草技术领域，具体涉及一种蚕蛹虫草的培养方法。
蚕蛹虫草是用野生天然蛹虫草（Cordycepsmilitaris）菌株孢子接种到蚕蛹上人工培育而成的虫菌结合体。
经分析检测证明：蚕蛹虫草所含虫草素、虫草酸、SOD等超过天然冬虫夏草、北虫草和榨蚕虫草，是一种真正能与野生冬虫夏草相媲美的虫草。
但传统的蚕蛹虫草的培养方法，培养时间长，产量和质量都有待提高。
本发明的目的在于提供一种蚕蛹虫草浓缩液的制作方法，其能提高蚕蛹虫草的产量和质量。
为实现上述目的，本发明的技术方案是设计一种蚕蛹虫草的培养方法，包括如下步骤：
1）挑选蚕蛹：挑选鲜活的蚕蛹；
2）接种：用注射器将液体菌种注射至蚕蛹体内；
3）暗培养：将接种后的蚕蛹放入恒温培养箱中避光培养，至蚕蛹表面长出白色菌丝；
4）光培养：对表面长出白色菌丝的蚕蛹进行光照培养，至菌丝由白色转为橘黄色。
优选的，步骤2）中，注射前，先用银针将蚕蛹刺穿多次。
优选的，所述银针的外径为2～3mm。
优选的，步骤2）中，液体菌种由接入培养基的蛹虫草菌种振荡培养而得。
优选的，按重量份计，所述培养基由以下组分组成：22～27份葡萄糖，2.7～4.3份乙醇，2.3～3.8份牛肉蛋白胨，0.7～1.4份过磷酸钙，3.6～5.7份玉米肽，0.8～1.3份硫酸镁，1.8～2.6份磷酸二氢钾，1.2～1.8份琼脂，0.1～0.3份反丁烯二酸，0.1～0.2份五水硫酸铜，1.5～2.1份酵母浸膏，100份蒸馏水。
优选的，所述振荡培养的温度控制在18～20℃，时间控制在15～20天。
优选的，步骤3）中，避光培养的温度控制在17～20℃，湿度控制在55%～60%，时间控制在6～9天。
优选的，步骤4）中，光照培养的温度控制在16～19℃，湿度控制在60%～75%，时间控制在20～25天。
本发明的优点和有益效果在于：提供一种蚕蛹虫草的培养方法，其能提高蚕蛹虫草的产量和质量。
本发明将蚕蛹刺穿多次，可缩短蚕蛹虫草的出草时间，同时也能增加出草量，还能提高蚕蛹虫草中虫草素、虫草酸、虫草多糖、SOD酶、喷司他丁等有效成分的含量。
本发明蚕蛹虫草的培养方法，以新鲜活体桑蚕蛹为培养基，不添加任何生长激素和抗生素，全程无污染，培养所得的蚕蛹虫草中含有虫草酸不低于4.77% ,粗蛋白不低于63.6%，赖氨酸不低于2.478%，明显高于其它蛹虫草，双向调节，阴阳互补，营养更全面，功效更显著。
下面结合实施例，对本发明的具体实施方式作进一步描述。以下实施例仅用于更加清楚地说明本发明的技术方案，而不能以此来限制本发明的保护范围。
本发明提供一种蚕蛹虫草的培养方法，包括如下步骤：
1）挑选蚕蛹：挑选鲜活的蚕蛹；
2）接种：用注射器将液体菌种注射至蚕蛹体内；
注射前，先用银针将蚕蛹刺穿多次（具体可为5～10次，且每次的刺穿角度都不同）；
银针的外径为2～3mm；
液体菌种由接入培养基的蛹虫草菌种振荡培养而得；
按重量份计，培养基由以下组分组成：22～27份葡萄糖，2.7～4.3份乙醇，2.3～3.8份牛肉蛋白胨，0.7～1.4份过磷酸钙，3.6～5.7份玉米肽，0.8～1.3份硫酸镁，1.8～2.6份磷酸二氢钾，1.2～1.8份琼脂，0.1～0.3份反丁烯二酸，0.1～0.2份五水硫酸铜，1.5～2.1份酵母浸膏，100份蒸馏水；
振荡培养的温度控制在18～20℃，时间控制在15～20天；
3）暗培养：将接种后的蚕蛹放入恒温培养箱中避光培养，至蚕蛹表面长出白色菌丝；
避光培养的温度控制在17～20℃，湿度控制在55%～60%，时间控制在6～9天；
4）光培养：对表面长出白色菌丝的蚕蛹进行光照培养，至菌丝由白色转为橘黄色；
光照培养的温度控制在16～19℃，湿度控制在60%～75%，时间控制在20～25天。
本发明将蚕蛹刺穿多次，可缩短蚕蛹虫草的出草时间，同时也能增加出草量，还能提高蚕蛹虫草中虫草素、虫草酸、虫草多糖、SOD酶、喷司他丁等有效成分的含量。
本发明蚕蛹虫草的培养方法，以新鲜活体桑蚕蛹为培养基，不添加任何生长激素和抗生素，全程无污染，培养所得的蚕蛹虫草中含有虫草酸不低于4.77% ,粗蛋白不低于63.6%，赖氨酸不低于2.478%，明显高于其它蛹虫草，双向调节，阴阳互补，营养更全面，功效更显著。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明技术原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。
Claims (8)
- 一种蚕蛹虫草的培养方法，其特征在于，包括如下步骤：1）挑选蚕蛹：挑选鲜活的蚕蛹；2）接种：用注射器将液体菌种注射至蚕蛹体内；3）暗培养：将接种后的蚕蛹放入恒温培养箱中避光培养，至蚕蛹表面长出白色菌丝；4）光培养：对表面长出白色菌丝的蚕蛹进行光照培养，至菌丝由白色转为橘黄色。
- 根据权利要求1所述的蚕蛹虫草的培养方法，其特征在于，步骤2）中，注射前，先用银针将蚕蛹刺穿多次。
- 根据权利要求2所述的蚕蛹虫草的培养方法，其特征在于，所述银针的外径为2～3mm。
- 根据权利要求3所述的蚕蛹虫草的培养方法，其特征在于，步骤2）中，液体菌种由接入培养基的蛹虫草菌种振荡培养而得。
- 根据权利要求4所述的蚕蛹虫草的培养方法，其特征在于，按重量份计，所述培养基由以下组分组成：22～27份葡萄糖，2.7～4.3份乙醇，2.3～3.8份牛肉蛋白胨，0.7～1.4份过磷酸钙，3.6～5.7份玉米肽，0.8～1.3份硫酸镁，1.8～2.6份磷酸二氢钾，1.2～1.8份琼脂，0.1～0.3份反丁烯二酸，0.1～0.2份五水硫酸铜，1.5～2.1份酵母浸膏，100份蒸馏水。
- 根据权利要求5所述的蚕蛹虫草的培养方法，其特征在于，所述振荡培养的温度控制在18～20℃，时间控制在15～20天。
- 根据权利要求6所述的蚕蛹虫草的培养方法，其特征在于，步骤3）中，避光培养的温度控制在17～20℃，湿度控制在55%～60%，时间控制在6～9天。
- 根据权利要求7所述的蚕蛹虫草的培养方法，其特征在于，步骤4）中，光照培养的温度控制在16～19℃，湿度控制在60%～75%，时间控制在20～25天。
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