CN102559735A - Method for producing nervonic acid by utilizing transgenic microalgae - Google Patents
Method for producing nervonic acid by utilizing transgenic microalgae Download PDFInfo
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- CN102559735A CN102559735A CN2011104466596A CN201110446659A CN102559735A CN 102559735 A CN102559735 A CN 102559735A CN 2011104466596 A CN2011104466596 A CN 2011104466596A CN 201110446659 A CN201110446659 A CN 201110446659A CN 102559735 A CN102559735 A CN 102559735A
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- acid
- selacholeic acid
- little algae
- selacholeic
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Abstract
The invention relates to a method for producing nervonic acid by utilizing transgenic microalgae. The method comprises the steps that the gene of a key enzyme, namely 3-ketoacyl CoA (coenzyme A) synthase, with high nervonic acid production capacity of fatty acid prolonged anabolism pathway in organism is transferred to a host microalgae cell, so that the microalgae is endowed with or enhanced in the nervonic acid synthesis capability; by cultivating the transgenic microalgae and promoting oil and fat accumulation of the microalgae, oil and fat with high content of nervonic acid can be extracted from the cultivated microalgae cells, and further, nervonic acid can be extracted from the oil and fat.
Description
Technical field
The present invention relates to a kind of working method of Selacholeic acid, be specifically related to a kind of method of producing Selacholeic acid, belong to Selacholeic acid genes produce technical field through the little algae of transgenic.
Background technology
Selacholeic acid, the chemical name nervonie acid is the long-chain monounsaturated fatty acids that a kind of ω 9 is, molecular formula is C
24H
46O
2, relative molecular mass is 366.6, is the white plates crystal at normal temperatures, can be dissolved in alcohol, and is water insoluble, 39 ~ 40 ℃ of fusing points.
Selacholeic acid is the important component in brain nervous cell and the nervous tissue, and in Mammals, Selacholeic acid is distributed widely in the sphingolipid of vertebrates tissue, and wherein Selacholeic acid combines with sheath amine alcohol through amido linkage.Be the white matter of animal brain and the important composition composition of peripheral nerve tissue.
Research shows; Selacholeic acid can promote damaged nerve tissue to repair and regeneration; The essential nutrients that is particularly brain cell, optic cell, peripheral nerve-cell growth of neurocyte, grows and keep again is to the active degree that improves cranial nerve, prevent that the cranial nerve aging from playing an important role.Selacholeic acid can be used as the adjuvant therapy medicaments or the supplementary of demyelinations such as multiple sclerosis, adrenal medullary atrophy disease, optic neuromyelitis disease, acute diffusivity myelencephalon inflammation and Zellweger syndromes.In addition, research shows also in the serum that the Selacholeic acid level is low has remarkable relation with sudden sensorineural deafness.
Selacholeic acid level and Psychiatric disorders are closely related.Psychiatric disorders, like schizophrenia, psychosis, attention disappearance etc., the level of Selacholeic acid is lower in patient's tissue and the blood.Selacholeic acid pair and fat relevant metabolic disease patient, accept hormone after climacteric and replace curer, alcoholics's etc. coronary artery disease danger prophylactic effect is arranged.
Selacholeic acid except have the treatment nervous system disorders, the effect that also has enhancing immunity.Research shows that Selacholeic acid is inhibited to the DNA polymerase beta, and Selacholeic acid can also suppress the active of HIV-1 ThermoScript II and show the dose-effect dependency.Even have and report that the Acertruncatum Bunge oil that is rich in Selacholeic acid has enhancing immunity, suppresses the effect of tumor growth and treatment AIDS therefore.
Because the important biomolecule of Selacholeic acid is active, Selacholeic acid production and have broad application prospect in the application aspect medicine and the nutrition.Add Selacholeic acid in the meals and can support myelinic synthetic and performance function in brain and the nervous tissue.The complementary diets that contains Selacholeic acid all has important meaning for heredity peroxisomal disease (like adrenoleukodystrophy, Zellweger ' s comprehensively demonstrate,proves) patient's lipid metabolism, multiple sclerosis patient's rehabilitation and the normal development of accepting the pronatis of infant formulas nutritious food.The comprehensive card of grease treatment Zellweger ' s with being rich in Selacholeic acid has also been obtained clinical efficacy.The level of Selacholeic acid and infant nutrition and development condition have dependency in the blood plasma; The Selacholeic acid level is than agalactous baby's height in breast-fed infant's red blood corpuscle; No matter the nutritionist is term birth or premature infant if recommending; If artificial feeding should be added Selacholeic acid, DPA (DPA) and DHA (docosahexenoic acid) in its milk-substitute.In order to increase the Selacholeic acid content of cow's milk, even the grease that has human to contain neural acid esters adds the Selacholeic acid content that improves milk in the milk cow feed to.Because Selacholeic acid is present in the whey, so these cow's milk that are rich in Selacholeic acid are particularly suitable for making the pleasant nutritional fortification sour milk of taste with special nutrition.Selacholeic acid and ARA (eicosatetraenoic acid), DHA be the same with linolenic acid to have the biological activity that promotes human health, can become the grease additive of protective foods.It is believed that Selacholeic acid will be the sour nutritional factor of biological activity fat that helps human health that will have the huge applications potentiality future, Selacholeic acid also will welcome the huge market opportunity with the grease that is rich in Selacholeic acid.
Yet; Up to the present; Worldwide also there is not large-scale production technology; A spot of Selacholeic acid mainly extracts from limited plant resources, perhaps the process of producing erucic acid from rapeseed oil, extracts the extremely low Selacholeic acid of content, and this can be through the situation formation sharp contrast of little algae scale operation with DHA's.So research and development have very important significance through the method that little algae produces Selacholeic acid.
Summary of the invention
The purpose of this invention is to provide a kind of method of producing Selacholeic acid through the little algae of transgenic.
The present invention is through transgenic technology, makes up the little algae algae kind with high yield Selacholeic acid ability, utilizes transgenic micro-algae large-scale cultivating and producing Selacholeic acid then.
Content of the present invention comprises; The gene that will have the fatty acid prolonging of biology of the high yield Selacholeic acid key enzyme (3-ketoacyl CoA synthase) in synthetic changes in host's microalgae cell; Little algae is had or strengthen the ability of synthetic Selacholeic acid; Thereby realization is through cultivating the little algae of transgenic and impel its accumulation grease, from little algae, extract grease or the further processing Selacholeic acid that is rich in Selacholeic acid.
The gene of 3-ketoacyl CoA synthase can come from the plant with high yield Selacholeic acid ability, like malania oleifera, acer truncatum, Longhairy Antenoron Herb, the mustard etc. of cracking rice.The host has stronger accumulation grease ability; And oleic acid content is more much higher than other fatty acid content in the fatty acid component in the grease; Or 22 monounsaturated fatty acids content below the carbon account for most little algae of unsaturated fatty acids total amount, as the new green alga of rich oil (
Neochloris oleoabundans) etc.
The present invention with respect to the beneficial effect of prior art is: at first, can improve the content of Selacholeic acid, the content that present people also do not grasp any little its Selacholeic acid of algae has commercial value, can obtain little algae algae kind of high yield Selacholeic acid through present technique; Secondly; Can solve the very low present situation of Selacholeic acid scarcity of resources thereby output; It is found that at present and have only a small amount of rare higher plant to have higher Selacholeic acid content, but because the harsh growth of these plant growing conditions is slow, so the output of Selacholeic acid can't improve; Micro algae growth is fast, culture condition is simple, can industriallization cultivate, so be hopeful to realize the high yield of Selacholeic acid through technology of the present invention; The 3rd, apply present technique and help protecting rare plant; The 4th; The technology of existing extraction Selacholeic acid is separation and Extraction from the very low vegetables oil of Selacholeic acid content mainly, and cost is very high, and the price of Selacholeic acid is very high; Be unfavorable for applying; Promote that technology of the present invention can reduce production costs, productivity gain, thereby can reduce product price, help the application of Selacholeic acid.
Embodiment
Through embodiment the present invention is explained further details below, these embodiment only are used for explaining the present invention, do not limit the scope of the invention.
Embodiment 1
Get the malania oleifera tissue, extract total RNA as masterplate, the mRNA with the primer RT-PCR that designs obtains 3-ketoacyl CoA synthase obtains total length with the RACE technology; Be connected to carrier, carrier injected the new chlorella cell of rich oil with particle gun, the screening transformant, transformant spreads cultivation; Collect microalgae cell, extract grease, hydrolysate oil; Analyze lipid acid with GC-MS and form, microalgae grease content 45% (pair cell dry weight) as a result, Selacholeic acid accounts for more than 40% of total fatty acids.
Embodiment 2
Get the acer truncatum tissue, extract total RNA, the mRNA with the primer RT-PCR that designs obtains 3-ketoacyl CoA synthase obtains total length with the RACE technology; Be connected to carrier, with utilizing Agrobacterium to change carrier over to chlorella cells, screen transformant, transformant spreads cultivation; Collect microalgae cell, extract grease, hydrolysate oil; Analyze lipid acid with GC-MS and form, microalgae grease content 30% (pair cell dry weight) as a result, Selacholeic acid accounts for more than 35% of total fatty acids.
Claims (6)
1. one kind is passed through the method that the little algae of transgenic produces Selacholeic acid, it is characterized in that:
With fatty acid prolonging in synthetic key enzyme---3-ketoacyl CoA synthase gene changes in host's microalgae cell; Little algae is had or strengthen the ability of synthetic Selacholeic acid; Thereby realize through cultivating this genetically modified little algae and impelling its accumulation grease; From the cultured microalgae cell, extract then and obtain the very high grease of Selacholeic acid content, from this grease, extract and obtain Selacholeic acid.
2. a kind of method of producing Selacholeic acid through the little algae of transgenic according to claim 1, it is characterized in that: the gene of described 3-ketoacyl CoA synthase comes from the biology with high yield Selacholeic acid ability.
3. a kind of method of producing Selacholeic acid through the little algae of transgenic according to claim 2 is characterized in that: described biology include but not limited to the plant malania oleifera (
Malania oleifera), acer truncatum (
Acer truncatum), Longhairy Antenoron Herb (
Lunaria annua), the mustard of cracking rice (
Cardamine graeca).
4. a kind of method of producing Selacholeic acid through the little algae of transgenic according to claim 1 is characterized in that: described host has accumulation grease ability to reach and surpass the little algae more than 10% that the cell fat content accounts for dried cell weight.
5. according to claim 1 or 4 described a kind of methods of producing Selacholeic acid through the little algae of transgenic; It is characterized in that: be used for the greasy fatty acid component of natural product as little algae of transformed host; Oleic acid content is higher than other any fatty acid content, or the ratio that the monounsaturated fatty acids content below 22 carbon accounts for the unsaturated fatty acids total amount is greater than 30%.
6. according to claim 1 or 5 described a kind of methods of producing Selacholeic acid, it is characterized in that through the little algae of transgenic: described host for but be not limited to the new green alga of rich oil (
Neochloris oleoabundans) and chlorella (
ChlorillaSp.).
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| CN2011104466596A CN102559735A (en) | 2011-12-28 | 2011-12-28 | Method for producing nervonic acid by utilizing transgenic microalgae |
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| CN2011104466596A CN102559735A (en) | 2011-12-28 | 2011-12-28 | Method for producing nervonic acid by utilizing transgenic microalgae |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104754954A (en) * | 2012-08-23 | 2015-07-01 | 特郎萨格以色列有限公司 | Transgenic microalgae and use thereof for oral delivery of proteins |
| CN106574255A (en) * | 2014-07-10 | 2017-04-19 | 泰拉瑞亚控股公司 | Ketoacyl acp synthase genes and uses thereof |
| US10480002B2 (en) | 2014-02-12 | 2019-11-19 | Transalgae Israel Ltd. | Algal based edible vaccines |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102220241A (en) * | 2011-05-06 | 2011-10-19 | 中国科学院青岛生物能源与过程研究所 | Cultivated fresh water microalgae and application thereof in production of biodiesel and nervonic acid |
-
2011
- 2011-12-28 CN CN2011104466596A patent/CN102559735A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102220241A (en) * | 2011-05-06 | 2011-10-19 | 中国科学院青岛生物能源与过程研究所 | Cultivated fresh water microalgae and application thereof in production of biodiesel and nervonic acid |
Non-Patent Citations (5)
| Title |
|---|
| 《Plant Mol Biol》 20091231 Yiming Guo et al. Increase in nervonic acid content in transformed yeast and transgenic plants by introduction of a Lunaria annua L. 3-ketoacyl-CoA synthase (KCS) gene 全文 1-6 , * |
| DAVID C. TAYLOR ET AL.: "Molecular cloning and characterization of a KCS gene from Cardamine graeca and its heterologous expression in Brassica oilseeds to engineer high nervonic acid oils for potential medical and industrial use", 《PLANT BIOTECHNOLOGY JOURNAL》, vol. 7, no. 9, 31 December 2009 (2009-12-31) * |
| YIMING GUO ET AL.: "Increase in nervonic acid content in transformed yeast and transgenic plants by introduction of a Lunaria annua L. 3-ketoacyl-CoA synthase (KCS) gene", 《PLANT MOL BIOL》, 31 December 2009 (2009-12-31) * |
| 王性炎 等: "神经酸研究现状及应用前景", 《中国油脂》, vol. 35, no. 3, 31 December 2010 (2010-12-31) * |
| 范勇 等: "利用微藻Mychonastes sp. HSO-3生产神经酸的研究初探", 《中国藻类学会第八次会员代表大会暨第十六次学术讨论会论文摘要集》, 11 November 2011 (2011-11-11), pages 2 - 4 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104754954A (en) * | 2012-08-23 | 2015-07-01 | 特郎萨格以色列有限公司 | Transgenic microalgae and use thereof for oral delivery of proteins |
| US9827280B2 (en) | 2012-08-23 | 2017-11-28 | Transalgae Israel Ltd. | Transgenic microalgae and use thereof for oral delivery of proteins |
| US11344590B2 (en) | 2012-08-23 | 2022-05-31 | Transalgae Israel Ltd. | Transgenic microalgae and use thereof for oral delivery of proteins |
| US10480002B2 (en) | 2014-02-12 | 2019-11-19 | Transalgae Israel Ltd. | Algal based edible vaccines |
| CN106574255A (en) * | 2014-07-10 | 2017-04-19 | 泰拉瑞亚控股公司 | Ketoacyl acp synthase genes and uses thereof |
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Application publication date: 20120711 |