CN1035627C - Preparation method for r-sub anacsthesia and biological products composed mainly of it - Google Patents
Preparation method for r-sub anacsthesia and biological products composed mainly of it Download PDFInfo
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- CN1035627C CN1035627C CN91106190A CN91106190A CN1035627C CN 1035627 C CN1035627 C CN 1035627C CN 91106190 A CN91106190 A CN 91106190A CN 91106190 A CN91106190 A CN 91106190A CN 1035627 C CN1035627 C CN 1035627C
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- linolenic acid
- linoleic acid
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Abstract
The present invention relates to a preparation method of r-linoleic acid and a biological preparation with the major ingredients of the r-linoleic acid, which belongs to the technical field of microorganism. The present invention is mainly characterized in that cunninghamella echinulata is adopted; a mutagenesis strain with high yield lipid and r-linoleic acid is obtained through microwave and ultraviolet mutation treatment; then, the biological preparation with the main ingredients of the r-linoleic acid is prepared through shake flask fermentation or tank (500 kL) fermentation; after being treated by a petroleum ether extraction method, r-linoleic acid grease is obtained. The biological preparation obtained by the present invention can be used in the technical fields of medicine, foodstuff, etc.
Description
The present invention system belongs to microorganism field, and promptly providing a kind of r-linolenic acid to reach with it is the preparation method and the biotechnological formulation of the biotechnological formulation of major ingredient.
R-linolenic acid (r-Linolenic acid) is a kind of C 18 unsaturated fatty acid, is called for short GLA.GLA is the precursor with highly bioactive prostaglandin(PG) (prostag landins), and it has the important physical effect in body.Therefore, over past ten years, the attention that the research of r-linolenic acid unsaturated fatty acids and application are subjected to countries in the world.Oenothera oil is present more welcome control high blood cholesterol drug, it is by extracting in the seed of Radix Oenotherae erythrosepalae, and its main component is a linolic acid, secondly is the r-linolenic acid, because linoleic acid content is higher than r-linolenic acid 7-8 doubly, linolenic physiological function of r-and drug effect have therefore been limited.In addition, since the influence of natural condition, the resource-constrained of root of Redsepal Eveningprimrose.Therefore, countries such as day, English have adopted microbial method to produce GLA, mainly utilize mortierella (Mortlerella), head mold (Rhlzopus) suitability for industrialized production r-linolenic acid, but the meta-bolites and the cell content that obtain can not used thereupon.
In order to solve the natural resources deficiency, the Microbial resources of development and use China, the invention provides a kind of r-linolenic acid and be the preparation method of the biotechnological formulation of major ingredient, and the tunning (comprising by product) of preparation is applied to fields such as medical science and food with it.
Principal character of the present invention is that it is made up of following operation: 1, separate the cunninghamella echinulata original strain from the wild-type strain that produces unsaturated fatty acids; 2, through microwave treatment 10-20 second, shake-flask culture is selected the high bacterial strain of r-linolenic acid output; 3, carry out mutagenic treatment with ultraviolet irradiation, select through shaking bottle again; 4,, under the temperature 26-30 ℃ of condition, be carbon source with glucose or sucrose, starch, molasses, acetate with strain culturing at PH6-5.5; With ammonium sulfate or urea, peptone is nitrogenous source, and contain vitamin H, in the matrix of VITAMIN and potassium magnesium inorganic salt oscillation and fermentation 96-144 hour or to adopt sucrose or the molasses of 6-10% be carbon source, C: N=30: 1, temperature is 26-28 ℃, and stirring velocity is that 200 rev/mins 500 liter jars fermented 140-150 hour; 5, filtration, washing, drying.Can obtain the r-linolenic acid by preparation method provided by the present invention is major ingredient, is equipped with the biotechnological formulation of secondary meta-bolites-polysaccharide, each seed amino acid, polyoxide dismutase and VITMAIN B1, B2 and the E of mutagenic fungi.In addition the product that aforesaid method obtained is handled with sherwood oil, can be extracted the linolenic grease of contained r-.
Original strain-cunninghamella echinulata of mentioning among the above-mentioned preparation method, a plant height that obtains after microwave and ultraviolet mutagenesis processing produces lipid and the higher mutagenic fungi of r-linolenic acid ability, this bacterial strain was cultivated 2 days under 28 ℃ of temperature in the murphy juice nutrient agar, colony diameter 3.0-4.0cm, mycelia is loose, the cotton wool shape, white, there is part white conidium to form, cultivated 4 days, colony diameter 4.0-6.0cm, mycelia is by leucismus lime look, and conidium is the lark or the colour of camel's hair, cultivated 6 days, mycelia is covered with incubator, is lark or drabon look, and sophisticated conidium is lark or tawny.Microscopically is observed the flourishing multi-branched of visible vegetative hyphae, diameter 10-20 micron, and conidiophore is upright, and the bubble bundle that the branch top forms has stigma, has formed the conidium of garden type or the very thin thorn of ellipse garden type band on the stigma, diameter 10-15 micron.
Below by embodiment in detail the linolenic preparation method of r-provided by the present invention and biotechnological formulation and preparation method thereof are described in detail.
The preparation method of the embodiment 1-linolenic shake flask fermentation of r-
(1) the r-linolenic acid produces the screening of bacterium
Take all factors into consideration according to the speed of growth, lipid and r-linolenic acid content, select the cunninghamella echinulata bacterial classification.
(2) microwave and ultraviolet radiation mutagenesis are handled
With the spore of the original strain that filters out, in microwave oven, handled for 15 seconds, shake-flask culture is measured lipid and r-linolenic acid content then, selects r-linolenic acid bacterial strain, uses uviolizing again, thereby has obtained high yield lipid, the linolenic bacterial strain of r-.
(3) shake flask fermentation
Above-mentioned bacterial strains is cultivated at glucose 4%, urea 0.12%, potassium hydrogen phosphate (KH
2PO
4) 0.05 sal epsom (MgSO
4.7H
2O) 0.025%, in the matrix of vitamins B, vitamin H trace, shaking culture is 4 days under 28-30 ℃ temperature.
(4) filtration, washing, drying.
(5) obtain the linolenic grease of r-with the Petroleum ether extraction method.
The preparation method of embodiment 2-r-linolenic acid 500 liter jars fermentation
(1) (2) step is identical with embodiment 1.
(3) adopting the sucrose (or molasses) with 10% be carbon source, C: N=30: 1, and temperature is 28 ℃, stirring velocity is that 200 rev/mins 500 liter jars ferment hour.
(4) (5) are also identical with embodiment 1.
One kind of embodiment 3-is biological agent of major ingredient and preparation method thereof with the r-linolenic acid
Different need not to obtain the linolenic grease of r-with the Petroleum ether extraction method present embodiment with embodiment 1,2, but will be after filtration, washing, dried product directly be used for as biotechnological formulation, its major ingredient is the r-linolenic acid, and all the other are multiple meta-bolites and cell content.The concrete composition of biotechnological formulation provided by the present invention is: grease yield 31-36%, and r-linolenic acid content 12-13%, dry cell weight 25-27 grams per liter contains 20 multiple amino acids, polysaccharide, many oxydase dismutase (SOD) and vitamins B in addition
1, B
2, metabolism product such as PP and E.
What the present invention will obtain is main component with the r-linolenic acid, and the biotechnological formulation that the above-mentioned multiple meta-bolites of proportioning is made has carried out the toxicity and the test of pesticide effectiveness of bacterial strain.With various dose abdominal injection and filling stomach mode, to 70 small white mouses, carried out acute toxicity test, by testing to sample continuously of a large amount of animals of 251 days (small white mouse and big white mouse), growth, survival and death, routine blood test, liver function, renal function index to animal detect, and prove no chronic toxicity and cumulative toxicity.
Carry out the test of pesticide effectiveness by isolated rabbit and animal being raised for a long time institute, its result proves, biotechnological formulation provided by the present invention has anticoagulant, reduce plasma viscosity, improve blood middle-high density lipoprotein, the effect of triglyceride reducing, whole blood viscosity and hematocrit in addition, also helps improving hemorheology status and changes atherosclerosis.
Moreover, biotechnological formulation provided by the present invention also is used to 30 patient's clinical observations 3 months, the result proves, but said preparation is triglyceride reducing and hematoblastic aggregation but also can make whole blood viscosity and cholesterol that tangible reduction is arranged not only, therefore, opened up new approach (seeing Appendix 1,2,3,4,5,6) for the control of cardiovascular and cerebrovascular diseases.
Can find out in sum, the linolenic preparation method of r-provided by the present invention than The advantages such as it is simple that prior art has technology, and the bacterial classification source is abundant, prepared biologic product can Be widely used in medical science, in the fields such as food.
Claims (1)
1. the preparation method of a gamma-linolenic acid is to adopt bacterial classification to send out technology pure, and products therefrom filtration, washing, dry extraction contain the grease of gamma-linolenic acid, it is characterized in that:
(1) bacterial classification is for producing the cunninghamella echinulata of gamma-linolenic acid;
(2) at PH:6-5.5, under the temperature 26-30 ℃ of condition, being carbon source, be nitrogenous source with urea, peptone with glucose or sucrose, starch, molasses, acetate with bacterial strain, to contain vitamin H, cultivated 96-144 hour in the matrix of VITAMIN and potassium magnesium inorganic salt;
(3) products therefrom filters, washing, and drying contains the grease of gamma-linolenic acid with Petroleum ether extraction.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN91106190A CN1035627C (en) | 1991-07-30 | 1991-07-30 | Preparation method for r-sub anacsthesia and biological products composed mainly of it |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN91106190A CN1035627C (en) | 1991-07-30 | 1991-07-30 | Preparation method for r-sub anacsthesia and biological products composed mainly of it |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1069070A CN1069070A (en) | 1993-02-17 |
| CN1035627C true CN1035627C (en) | 1997-08-13 |
Family
ID=4907625
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN91106190A Expired - Fee Related CN1035627C (en) | 1991-07-30 | 1991-07-30 | Preparation method for r-sub anacsthesia and biological products composed mainly of it |
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Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100348514C (en) * | 2005-03-29 | 2007-11-14 | 华中科技大学 | Deep-processing method of using starch watewater |
| CN1329519C (en) * | 2006-03-29 | 2007-08-01 | 王景恒 | Gamma-linolenic acid dry microbial powder enriched with organic celenium and its producing process |
| CN102676594A (en) * | 2012-05-25 | 2012-09-19 | 厦门大学 | Preparation method of gamma-linolenic acid |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0252716A2 (en) * | 1986-07-08 | 1988-01-13 | Suntory Limited | Process for production of Bishomo- Gamma-Linolenic acid and eicosapentaenoic acid |
| EP0322227A2 (en) * | 1987-12-21 | 1989-06-28 | Suntory Limited | Process for production of bishomo-gamma-linolenic acid |
| EP0399494A1 (en) * | 1989-05-24 | 1990-11-28 | Idemitsu Petrochemical Co. Ltd. | Process for production of dihomo-gamma-linolenic acid and inhibitor for unsaturation reaction at delta 5-position of fatty acid |
-
1991
- 1991-07-30 CN CN91106190A patent/CN1035627C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0252716A2 (en) * | 1986-07-08 | 1988-01-13 | Suntory Limited | Process for production of Bishomo- Gamma-Linolenic acid and eicosapentaenoic acid |
| EP0322227A2 (en) * | 1987-12-21 | 1989-06-28 | Suntory Limited | Process for production of bishomo-gamma-linolenic acid |
| US4916066A (en) * | 1987-12-21 | 1990-04-10 | Suntory Limited | Process for production of bishomo-gamma-linolenic acid |
| EP0399494A1 (en) * | 1989-05-24 | 1990-11-28 | Idemitsu Petrochemical Co. Ltd. | Process for production of dihomo-gamma-linolenic acid and inhibitor for unsaturation reaction at delta 5-position of fatty acid |
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| Publication number | Publication date |
|---|---|
| CN1069070A (en) | 1993-02-17 |
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