CN103627770A - Method used for biotransformation of pseudo-ginseng using two paecilomyces funguses - Google Patents
Method used for biotransformation of pseudo-ginseng using two paecilomyces funguses Download PDFInfo
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- CN103627770A CN103627770A CN201210301911.9A CN201210301911A CN103627770A CN 103627770 A CN103627770 A CN 103627770A CN 201210301911 A CN201210301911 A CN 201210301911A CN 103627770 A CN103627770 A CN 103627770A
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Abstract
The invention relates to a method used for biotransformation of pseudo-ginseng using two paecilomyces funguses. The method mainly comprises following steps: (1) preparation of Paecilomyce shepiali Chen and Dai, and Paecilomyces militaris Liang; (2) preparation of a pseudo-ginseng culture medium; (3) setting of fermentation cultivation conditions; and (4) product postprocessing and detection of main functional ingredients. According to the method, pseudo-ginseng is directly taken as a fermentation substrate, and medicinal components of both pseudo-ginseng and medicinal cordyceps fungi are obtained via one-step fermentation, so that the culturing product possess both the effects of pseudo-ginseng and the medicinal fungi.
Description
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Technical field
The invention belongs to microbial technology field, relate to two kinds of paecilomyces fungies and pseudo-ginseng is carried out to the method for bio-transformation.
Background technology
Pseudo-ginseng [
panax notoginseng(Burk.) F. H. Chen] be the traditional medicinal plant of Araliaceae Panax per nnial herb ，Shi China.Main active ingredient be Radix Notoginseng total arasaponins (
panax notoginsengsaponins, PNS).Radix Notoginseng total arasaponins is remarkable in hypoxia tolerance, pharmacological action anti-ageing, that improve the aspects such as immunity of organisms, and total saponins and partial monosomy saponin(e all have better activity at blood system, cardio-cerebrovascular, neural system, substance metabolism and anti-inflammatory, the aspect such as antitumor.Especially rare saponin(e shows good pharmacologically active at treatment cardiovascular and cerebrovascular diseases, anti-tumor aspect, as Ginsenoside Rd is unique to effects such as cardiovascular and cerebrovascular, neural system, immunity systems, is a kind of very promising drug candidate.Research shows: natural content of ginsenoside is low, and molecular structure is not best active condition, can reduce the glycosyl number in ginsenoside structure by bio-transformation, the saponin(es such as the Rg1 that in natural saponin(e, content is higher, Rb1, R1, Rc are converted into the very micro-rare saponin(e of content: Rd, Rh1, F1, F2, C-K etc., can improve its curative effect and physiologically active.
Peacilomyce hepiahi (
paecilomyces hepialichen & Dai) and pupa draw up mould (
paecilomyces militarisliang) be two kinds of important medicinal Cordyceps fungus, in this fungus fermentation products, contain that to take cordycepin and Chinese caterpillar fungus adenosine be main medicinal ingredients, there is the effect of antitumor, antibacterial, antiviral, immunomodulatory, removing free radical.
The present invention be take pseudo-ginseng as main medium composition, adopt two kinds of medicinal fungi peacilomyce hepiahis (
paecilomyces hepialichen & Dai) and pupa draw up mould (
paecilomyces militarisliang) pseudo-ginseng is carried out to bio-transformation, obtain medicinal rare ginsenoside, cordycepin and Chinese caterpillar fungus adenosine simultaneously, culture has pseudo-ginseng drug effect and fungi drug effect concurrently, has improved treatment and health-care effect.
Summary of the invention
The invention provides a kind of medicinal fungi fermentation pseudo-ginseng that utilizes, obtain the method for fungi and pseudo-ginseng active pharmaceutical ingredients simultaneously.
The operation steps of the method is as follows:
(1) bacterial classification preparation: by peacilomyce hepiahi (
paecilomyces hepialichen & Dai) and pupa draw up mould (
paecilomyces militarisliang) be inoculated into respectively PPDA slant medium, in 26 ℃ of constant temperature culture 7 days;
(2) pseudo-ginseng substratum: accurately take Radix Notoginseng powder that 1 g crosses 60 mesh sieves in 100 mL triangular flasks, add 0.2 mL inorganic salt: 10 %KH
2pO
4with 5 %MgSO
4, 20 mL water, natural pH, 121 ℃ of sterilizing 30 min;
(3) liquid fermentation and culture: by bacterial classification access step (2) the pseudo-ginseng substratum preparing in step (1), in 10-30 ℃, 120 r/min cultivate 4-16 d;
(4) liquid fermentate aftertreatment: by cultured liquid fermentate suction filtration in step (3), with 10 mL pure water, clean filter residue, merging filtrate, centrifugal 30 min of 4000 r/min, it is supernatant treatment solution that supernatant liquor adds pure water to be settled to 30mL.Get 10 mL supernatant treatment solutions and add equal-volume water-saturated n-butanol ultrasonic extraction 1 h, after stratification, collect 60 ℃ of upper strata propyl carbinol phases and revolve steaming to dry, with 5 mL 70% dissolve with methanol, adopt HPLC method to detect its saponin content.Get 20 mL supernatant treatment solutions and dry, get 0.1 g and should dry sample and add 1 mL 20% dissolve with methanol, employing HPLC method detects its cordycepin and Chinese caterpillar fungus adenosine content.
Key point of the present invention is: use medicinal Cordyceps fungus to carry out bio-transformation to pseudo-ginseng, one time fermentation process obtains fungi effective component and pseudo-ginseng effective component simultaneously, makes culture have the double effects of pseudo-ginseng and medicinal fungi concurrently.
Embodiment
Following examples are used for illustrating the present invention, but are not limitations of the present invention.
Embodiment 1: the impact of different culture temperature on tunning effective constituent kind and content
(1) bacterial classification preparation: by peacilomyce hepiahi (
paecilomyces hepialichen & Dai) and pupa draw up mould (
paecilomyces militarisliang) be inoculated into respectively PPDA slant medium, in 26 ℃ of constant temperature culture 7 days;
(2) pseudo-ginseng substratum: accurately take Radix Notoginseng powder that 1 g crosses 60 mesh sieves in 100 mL triangular flasks, add 0.2 mL inorganic salt: 10 %KH
2pO
4with 5 %MgSO
4, 20 mL water, natural pH, 121 ℃ of sterilizing 30 min;
(3) liquid fermentation and culture: by bacterial classification access step (2) the pseudo-ginseng substratum preparing in step (1), in 10 ℃ and 30 ℃, 120 r/min cultivate 10 d;
(4) liquid fermentate aftertreatment: by cultured liquid fermentate suction filtration in step (3), with 10 mL pure water, clean filter residue, merging filtrate, centrifugal 30 min of 4000 r/min, it is supernatant treatment solution that supernatant liquor adds pure water to be settled to 30 mL.Get 10 mL supernatant treatment solutions and add equal-volume water-saturated n-butanol ultrasonic extraction 1 h, after stratification, collect 60 ℃ of upper strata propyl carbinol phases and revolve steaming to dry, with 5 mL 70% dissolve with methanol, adopt HPLC method to detect its saponin content.Get 20 mL supernatant treatment solutions and dry, get 0.1 g and should dry sample and add 1 mL 20% dissolve with methanol, employing HPLC method detects its cordycepin and Chinese caterpillar fungus adenosine content.It the results are shown in Table 1.
The impact of the different culture temperature of table 1 on tunning effective constituent kind and content
Embodiment 2: the impact of different incubation times on tunning effective constituent kind and content
(1) bacterial classification preparation: by peacilomyce hepiahi (
paecilomyces hepialichen & Dai) and pupa draw up mould (
paecilomyces militarisliang) be inoculated into respectively PPDA slant medium, in 26 ℃ of constant temperature culture 7 days;
(2) pseudo-ginseng substratum: accurately take Radix Notoginseng powder that 1 g crosses 60 mesh sieves in 100 mL triangular flasks, add 0.2 mL inorganic salt: 10 %KH
2pO
4with 5 %MgSO
4, 20 mL water, natural pH, 121 ℃ of sterilizing 30 min;
(3) liquid fermentation and culture: by bacterial classification access step (2) the pseudo-ginseng substratum preparing in step (1), in 25 ℃, 120 r/min cultivate 4 d and 16 d;
(4) liquid fermentate aftertreatment: by cultured liquid fermentate suction filtration in step (3), with 10 mL pure water, clean filter residue, merging filtrate, centrifugal 30 min of 4000 r/min, it is supernatant treatment solution that supernatant liquor adds pure water to be settled to 30 mL.Get 10 mL supernatant treatment solutions and add equal-volume water-saturated n-butanol ultrasonic extraction 1 h, after stratification, collect 60 ℃ of upper strata propyl carbinol phases and revolve steaming to dry, with 5 mL 70% dissolve with methanol, adopt HPLC method to detect its saponin content.Get 20 mL supernatant treatment solutions and dry, get 0.1 g and should dry sample and add 1 mL 20% dissolve with methanol, employing HPLC method detects its cordycepin and Chinese caterpillar fungus adenosine content.It the results are shown in Table 2.
The impact of the different incubation times of table 2 on tunning effective constituent kind and content
Claims (1)
1. two kinds of paecilomyces fungies carry out the method for bio-transformation to pseudo-ginseng, it is characterized in that the method comprises:
(1) producing bacterial strain is peacilomyce hepiahi
paecilomyces hepialichen & Dai and pupa are drawn up mould
paecilomyces militarisliang;
(2) bacterial classification preparation: peacilomyce hepiahi and pupa are drawn up to mould and be inoculated into respectively PPDA slant medium, in 26 ℃ of constant temperature culture 7 days;
(3) pseudo-ginseng substratum: accurately take Radix Notoginseng powder that 1 g crosses 60 mesh sieves in 100 mL triangular flasks, add 0.2 mL inorganic salt: 10 %KH
2pO
4with 5 %MgSO
4, 20 mL water, natural pH, 121 ℃ of sterilizing 30 min;
(4) liquid fermentation and culture: by the bacterial classification access pseudo-ginseng substratum preparing, in 10-30 ℃, 120 r/min cultivate 4-16 d;
(5) liquid fermentate aftertreatment: by cultured liquid fermentate suction filtration, with 10 mL pure water, clean filter residue, merging filtrate, centrifugal 30 min of 4000 r/min, it is supernatant treatment solution that supernatant liquor adds pure water to be settled to 30 mL, get 10 mL supernatant treatment solutions and add equal-volume water-saturated n-butanol ultrasonic extraction 1 h, after stratification, collect 60 ℃ of upper strata propyl carbinol phases and revolve steaming to dry, with 5 mL70% dissolve with methanol, adopt HPLC method to detect its saponin content, getting 20 mL supernatant treatment solutions dries, getting 0.1 g should dry sample and added 1 mL 20% dissolve with methanol, adopt HPLC method to detect its cordycepin and Chinese caterpillar fungus adenosine content.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108676730A (en) * | 2018-05-25 | 2018-10-19 | 江西国药有限责任公司 | A kind of fermentation manufacturing technique of Paecilomyces hepiali chen Cs-4 |
| CN108865895A (en) * | 2018-06-15 | 2018-11-23 | 浙江工业大学 | Paecilomyces hepiali chen ZJB18001 and its application |
| CN109223835A (en) * | 2018-09-28 | 2019-01-18 | 贵州师范大学 | A kind of method of cordyceps sinensis Relative Fungi common fermentation production Radix Astragali Radix Notoginseng mycoplasma |
| CN110592161A (en) * | 2019-09-24 | 2019-12-20 | 长春工业大学 | Preparation method of polysaccharide, health-care oral liquid and preparation method |
| CN113151362A (en) * | 2021-03-26 | 2021-07-23 | 文山学院 | Preparation method of pseudo-ginseng solid fermentation product |
Citations (1)
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| CN102091109A (en) * | 2010-09-25 | 2011-06-15 | 郭景龙 | Medical composite for treating cardiovascular and cerebrovascular disease |
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| CN102091109A (en) * | 2010-09-25 | 2011-06-15 | 郭景龙 | Medical composite for treating cardiovascular and cerebrovascular disease |
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| 周超群等: "人参皂苷Rd高产突变株Paecilomuyces bainer sp229-7的生长特性和生物转化", 《复旦学报（医学版）》 * |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108676730A (en) * | 2018-05-25 | 2018-10-19 | 江西国药有限责任公司 | A kind of fermentation manufacturing technique of Paecilomyces hepiali chen Cs-4 |
| CN108676730B (en) * | 2018-05-25 | 2021-02-19 | 江西国药有限责任公司 | Fermentation production process of paecilomyces hepiali Cs-4 |
| CN108865895A (en) * | 2018-06-15 | 2018-11-23 | 浙江工业大学 | Paecilomyces hepiali chen ZJB18001 and its application |
| CN108865895B (en) * | 2018-06-15 | 2020-12-25 | 浙江工业大学 | Paecilomyces hepiali ZJB18001 and application thereof |
| CN109223835A (en) * | 2018-09-28 | 2019-01-18 | 贵州师范大学 | A kind of method of cordyceps sinensis Relative Fungi common fermentation production Radix Astragali Radix Notoginseng mycoplasma |
| CN110592161A (en) * | 2019-09-24 | 2019-12-20 | 长春工业大学 | Preparation method of polysaccharide, health-care oral liquid and preparation method |
| CN113151362A (en) * | 2021-03-26 | 2021-07-23 | 文山学院 | Preparation method of pseudo-ginseng solid fermentation product |
| CN113151362B (en) * | 2021-03-26 | 2022-11-01 | 文山学院 | Preparation method of pseudo-ginseng solid fermentation product |
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Application publication date: 20140312 |