CN1082089C - Fermentation technology of xylaria nigripes - Google Patents

Fermentation technology of xylaria nigripes Download PDF

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Publication number
CN1082089C
CN1082089C CN96106492A CN96106492A CN1082089C CN 1082089 C CN1082089 C CN 1082089C CN 96106492 A CN96106492 A CN 96106492A CN 96106492 A CN96106492 A CN 96106492A CN 1082089 C CN1082089 C CN 1082089C
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powder
tank
xylaria nigripes
seed
jar
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CN1157852A (en
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俞永信
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Zhou Lili
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Abstract

The present invention discloses xylaria nigripes sacc which can be cultured under manual conditions, and a method for carrying out large-scale industrialization production. Mycelia which are completely similar to wild bacteria can be produced by a manual culture medium under the culture conditions.

Description

Fermentation technology of Xylaria nigripes
The present invention is about the deep layer fermenting process of Xylaria nigripes.
Xylaria nigripes is a kind of medicinal fungi of preciousness, mainly is distributed in ground such as Sichuan, Yunnan, is famous qi-invigorating herb.It parasitizes on the black wing termitary of depleted, and apart from ground 2-3 rice, resource is few.Its extract has anti-tumor activity, strengthens cell and humoral immune reaction, and can promote to grow, and its medicinal products is wild bacterium at present, because natural resources is very rare, still can not satisfy needs of society.In order to discover and use this useful medicinal fungus, need carry out artificial culture, to satisfy the human needs that give protection against cancer, control cancer, health care.
The inventor is through groping repeatedly for many years, from numerous wild strains, be separated to a strain can be on specific artificial medium the bacterial strain of growth and breeding, and developed the technology that adopts artificial this bacterial strain of submerged fermentation, obtain the mycelium with the identical pharmacological action of wild bacterium tool, and obtained to contain the fermented liquid of the meta-bolites that abundant nutrition is worth.
The bacterial strain Xylaria nigripes that the present invention is separated to (Xyiaria nigripes (KL) Sacc) has been preserved in Chinese typical culture collection center, and preserving number is: CCTCC.NO93028.
One, the continuous fermentation process of charcoal angle bacterium
(1) substratum:
Solid culture: glucose 40 grams, yeast powder 20 grams, peptone 5 grams, MgSO 40.5 gram, agar 20 grams, water 1000 grams after substratum is sterilized routinely, insert the bacterial classification that the present invention is separated to, and cultivate 4 days under 24-26 ℃ of constant temperature, grow the living mycelia of greyish white color base on the inclined-plane, continue to cultivate, and grow dense aerial hyphae on the inclined-plane.
Liquid culture based formulas: Semen Maydis powder 60 grams, analysis for soybean powder 15 grams, yeast powder 15 grams, dried silkworm chrysalis meal 5 grams, MgSO 40.3 gram, pH=6.5, add water 100 grams, configuration is in proportion poured triangular flask into and is cultivated 1/3 of fiduciary point capacity after the sterilization, insert inclined-plane, front bacterium piece, under 26 ℃ of constant temperatures, shaking culture 48 hours, quite dense bacterium ball appears in the cultivation initial stage in the substratum of triangular flask, and later bacterium ball density increases rapidly, through the microscopy no bacteria pollution, promptly can be used as the seeding tank seed.
(2) fermentative production
1. seed tank culture
Substratum: Semen Maydis powder 60kg, yeast powder 15kg, analysis for soybean powder 20kg, dried silkworm chrysalis meal 5kg, MgSO 40.5kg, water 1000kg, pH6.5, culture condition: temperature 26-28 ℃, air flow is 1: 1, and do not stir early stage, and the rotating speed of later stage with per minute 100 times stirs seed tank culture 60 hours.
During the seeding tank enlarged culturing, carry out once sugar after the 8th hour, nitrogen detects and microscopy, checks once every 8 hours later on, moves into fermentor tank expansion behind the microscopy no bacteria pollution and produces.
2. fermentor cultivation
Substratum: Semen Maydis powder 80kg, yeast powder 20kg, dried silkworm chrysalis meal 10kg, analysis for soybean powder 20kg, MgSO 40.5kg, KH 2PO 41.0kg, water 1000kg, pH6.5.
Behind the preparation substratum, sterilization routinely (125 ℃ continue 20 minutes) is cooled to 26 ℃, the inoculation after the immigration seed tank culture, and inoculum size is 10%, air flow 1: 1.
Stirring fells out of use, rotating speed is per minute 100 times after 16 hours, fermentation time is 84 hours, leavening temperature is 26 ± 2 ℃, gets fermented liquid every 8 hours after the 16th hour, detects reducing sugar and nitrogen, whether microscopic examination has bacterial contamination, fermenting, sugar is almost utilized fully after 84 hours, and the sugar of detection is below 0.1%, and pH drops to about 5.5.Fermented liquid is also thicker, and illustrating ferments finishes, and can go out jar, and jar interior surplus 15% fermented liquid, is pressed into interior continuation of fermentor tank and ferments the cooling nutrient solution after the sterilization as the seed of next jar fermentation.
3. aftertreatment
Fermented liquid is pressed into plate-and-frame filter press, separate fermentation liquid and mycelium, mycelium drying, pulverizing, standby.
Fermented liquid includes a large amount of meta-bolitess and comprises polysaccharide, VITAMIN, amino acid utilizes the great disparity of above-mentioned product and molecular weight water, by film separate, dry, standby.
Two, the composition of Xylaria nigripes
1. polysaccharide: contain polysaccharide such as N.F,USP MANNITOL, rhamnosyl, lactose and reach 9.23%.
2. amino acid: charcoal angle bacterium has 18 seed amino acids, and total amount is 9.001%.
3. micro-: trace elements such as Fe, Mn, Sn, Cr, Ni, Mo, Cu, Zn, Co, Sr, B, Ca, K, Na, P.
4. VITAMIN: Vd27.4ppm, Ve21.7ppm, Vk6.8ppm, beta carotene 1.4ppm.
Three, toxicological test
Acute toxicity test shows, the actual nontoxicity of this product.Use 50mg, 200mg, 1000mg/kg jar stomach, the trimestral long term toxicity test of administration is to the heart, liver, stomach, kidney vitals and the equal non-toxic reaction of hemopoietic system." teratogenesis, carcinogenic, mutagenesis " test shows that this product is all negative, no teratogenesis, carcinogenic, mutagenesis influence.
Four, pharmacological testing
1. raise immunity suppresses tumour
Test shows, this product is remarkable activating macrophage (MQ) in vivo, thereby after strengthening that its cytotoxic effect shows scavenger cell and tumour cell is external and hatching jointly, the propagation of tumour cell obviously is subjected to press down, inhibiting rate can reach more than 85%, in the supernatant liquor that this product and scavenger cell effect were gathered in the crops after 4 hours, contain the activity of lymphokine IL-1 (interleukin 1), show that it can improve the thymus gland propagation that ConA stimulates.In the immunity system of body, scavenger cell occupies an important position, its auxiliary lymphocyte responses, IL-1 that the activatory scavenger cell is secreted and Interferon, rabbit, it is again the stimulating factor of proliferative T cell NK cytoactive, scavenger cell can produce specificity and non-specific killer factor to tumour cell in tumour immunity, this product is activating macrophage in vivo, thereby tumour cell is produced cytotoxic effect, and confirm that normal mice administration and the administration of band knurl mouse all can promote the cytotoxic effect of scavenger cell, suppress the propagation of tumour cell.
2. cerebral protection
The mouse broken end is panted to test and is shown, 250mg, and the 500mg/kg subcutaneous injection all has tangible cerebral protection, and the dysmnesia that the mouse electroshock is caused have significant protective effect.
3. promote growth
50-200mg/kg irritates stomach.From first week, rat body weight obviously increases, and the grown promoter action of this product to young mouse is described.
4. protect effects such as liver
Can significantly reduce SGPT.The 50mg-200mg/kg administration after 1.5 months oxyphorase obviously improve, and significantly reduce serum urea nitrogen.
Five. clinical verification
Clinical verification shows, this product is to dysmenorrhes, menoxenia, and multiple gynaecological disease such as menorrhagia is very few and man's prostatitis have obvious curative effects.

Claims (5)

1. Xylaria nigripes CCTCC.NO93028.
2. the continuous ferment process of Xylaria nigripes, after it is characterized in that Xylaria nigripes CCTCC.NO93028 cultivated in seeding tank, the inoculum size with 10% inserts fermentor cultivation.
3. according to the described continuous ferment process of claim 2, it is characterized in that the seed tank culture condition is temperature 26-28 ℃, air flow is 1: 1,26 ± 2 ℃ of fermentor cultivation condition leavening temperatures, and the rotating speed that ferments after 16 hours with per minute 100 times stirs.
4. according to the described continuous ferment process of claim 2, it is characterized in that the seed tank culture base: Semen Maydis powder 62kg, yeast powder 15kg, analysis for soybean powder 20kg, dried silkworm chrysalis meal 5kg, MgSO 40.5kg, water 1000kg, pH6.5, fermention medium: Semen Maydis powder 80kg, yeast powder 20kg, dried silkworm chrysalis meal 10kg, analysis for soybean powder kg, MgSO 40.5kg, KH 2PO 41.0kg, water 1000kg, pH6.5.
5. according to the described continuous ferment process of claim 3, it is characterized in that to go out jar after the ferment tank 84 hours.Surplus 15% fermented liquid, is pressed into and continues fermentation in the fermentor tank sterile medium as the seed of next jar fermentation in jar.
CN96106492A 1996-08-16 1996-08-16 Fermentation technology of xylaria nigripes Expired - Fee Related CN1082089C (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN96106492A CN1082089C (en) 1996-08-16 1996-08-16 Fermentation technology of xylaria nigripes

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Application Number Priority Date Filing Date Title
CN96106492A CN1082089C (en) 1996-08-16 1996-08-16 Fermentation technology of xylaria nigripes

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CN1157852A CN1157852A (en) 1997-08-27
CN1082089C true CN1082089C (en) 2002-04-03

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Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1699549B (en) * 2004-05-22 2011-06-08 朱志熊 Xylaria nigripes and rejuvenation, slant culture and variation degradation prevention technology
CN103999691B (en) * 2014-05-30 2016-03-30 杭州桐君堂生物科技有限公司 A kind of method simultaneously obtaining rich selenium solid fermentation mycelium and rich selenium crow spirit ginseng sporophore
TWI619499B (en) * 2015-07-17 2018-04-01 靜宜大學 Neuroprotection application of xylaria nigripes mycelium extract
CN110241145B (en) * 2019-06-27 2023-04-25 安阳师范学院 Fermentation preparation method of mannitol

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1104249A (en) * 1994-08-25 1995-06-28 俞永信 Deep fermentation technology for cordyceps sinensis sacc

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1104249A (en) * 1994-08-25 1995-06-28 俞永信 Deep fermentation technology for cordyceps sinensis sacc

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