MD4122C1 - Strain of Spirulina platensis (Nordst) Geitl alga as a source of biologically active substances - Google Patents

Strain of Spirulina platensis (Nordst) Geitl alga as a source of biologically active substances Download PDF

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MD4122C1
MD4122C1 MDA20110016A MD20110016A MD4122C1 MD 4122 C1 MD4122 C1 MD 4122C1 MD A20110016 A MDA20110016 A MD A20110016A MD 20110016 A MD20110016 A MD 20110016A MD 4122 C1 MD4122 C1 MD 4122C1
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geitl
nordst
alga
biologically active
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MD4122B1 (en
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Валериу РУДИК
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Валериу РУДИК
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Abstract

The invention relates to biotechnology and can be used for production of biologically active substances.Strain of Spirulina platensis (Nordst) Geitl alga is deposited with the National Collection of Nonpathogenic Microorganisms of the Republic of Moldova of the Institute of Microbiology and Biotechnology of the ASM under the number CNMN-CB-11 and can be used as a source of glutamic and γ - linoleic acids, sulphated polysaccharides, phosphatidylinositol and phosphatidylcholine.The result is to increase the productivity of alga and the quantity of glutamic and γ - linoleic acids, sulphated polysaccharides, phosphatidylinositol and phosphatidylcholine.

Description

Invenţia se referă la biotehnologie şi poate fi utilizată pentru obţinerea substanţelor biologic active. The invention refers to biotechnology and can be used to obtain biologically active substances.

Biomasa de spirulină oferă un spectru deplin al aminoacizilor, prezenţi atât în stare liberă, cât şi legată. Un interes practic major îl prezintă aminoacizii imunoactivi, care joacă un rol esenţial în procesele de protecţie a organismului. Pe lângă cele menţionate, acidul glutamic mai posedă şi funcţia de neuroprotecţie. Polizaharidele sulfatate prezente în biomasa de spirulină manifestă activitate antitumorală, antivirală şi imunostimulatoare. Fracţia fosfolipidelor esenţiale include şi fosfatidilinozitolul, care este recunoscut în calitate de normolipemiant şi cardioprotector. Un alt component bioactiv din spirulină este acidul gras polinesaturat γ-linolenic, care se caracterizează prin acţiune antieterogenă pronunţată. Spirulina biomass offers a full spectrum of amino acids, present both in free and bound state. Of major practical interest are the immunoactive amino acids, which play an essential role in the body's protection processes. In addition to the mentioned, glutamic acid also has the function of neuroprotection. The sulfated polysaccharides present in spirulina biomass exhibit antitumor, antiviral and immunostimulating activity. The fraction of essential phospholipids also includes phosphatidylinositol, which is recognized as a normolipemiant and cardioprotector. Another bioactive component of spirulina is the γ-linolenic polyunsaturated fatty acid, which is characterized by pronounced antiheterogeneous action.

Este cunoscută tulpina Spirulina platensis (Nordst) Geitl, CALU-835. Biomasa absolut uscată a acestei tulpini conţine 65…70% proteine, 10…15% hidraţi de carbon, 3…5% lipide, 0,42…0,44% β-carotină, 80…100 mg/% acid ascorbic, 30…35 mg/% tocoferol, vitamine din grupa B, 0,36% acid glutamic, fitohormoni şi alte substanţe biologic active [1]. The strain Spirulina platensis (Nordst) Geitl, CALU-835, is known. The absolutely dry biomass of this strain contains 65...70% proteins, 10...15% carbohydrates, 3...5% lipids, 0.42...0.44% β-carotene, 80...100 mg/% ascorbic acid, 30... 35 mg/% tocopherol, group B vitamins, 0.36% glutamic acid, phytohormones and other biologically active substances [1].

Mai este cunoscută tulpina Spirulina platensis (Nordst) Geitl, CNM-CB-01, cu biomasa absolut uscată, care conţine: 1,3…3,3% aminoacizi liberi, 5,5…6,5% oligopeptide, 62…70% proteine, 9…11% ficobiliproteide, 10…15% polizaharide, 3…5% lipide, 0,42…0,44% β-carotină, 30…35 mg/% tocoferol, 80…100 mg/% acid ascorbic, vitaminele grupei B, 1,0…1,2% acid γ-linolenic; 6,0…6,3% acid glutamic, fitohormoni şi alte substanţe bioactive [2]. Also known is the strain Spirulina platensis (Nordst) Geitl, CNM-CB-01, with absolutely dry biomass, which contains: 1.3...3.3% free amino acids, 5.5...6.5% oligopeptides, 62...70% proteins, 9...11% phycobiliproteins, 10...15% polysaccharides, 3...5% lipids, 0.42...0.44% β-carotene, 30...35 mg/% tocopherol, 80...100 mg/% ascorbic acid, vitamins group B, 1.0...1.2% γ-linolenic acid; 6.0...6.3% glutamic acid, phytohormones and other bioactive substances [2].

Dezavantajele tulpinilor menţionate mai sus constau în conţinutul redus al compuşilor bioactivi de interes major, aşa ca acidul glutamic, acidul γ-linolenic, fosfatidilinozitolul şi polizaharidele sulfatate. Valorile acestor indici sunt date în tabel. The disadvantages of the strains mentioned above consist in the low content of bioactive compounds of major interest, such as glutamic acid, γ-linolenic acid, phosphatidylinositol and sulfated polysaccharides. The values of these indices are given in the table.

Tabel Table

Conţinutul unor substanţe bioactive în biomasa a două tulpini de spirulină, % biomasă absolut uscată The content of some bioactive substances in the biomass of two spirulina strains, % absolute dry biomass

Tulpina Acidul glutamic Acidul γ-linolenic Fosfatidilinozitolul Fosfatidil-colină Polizaharide sulfatate Spirulina platensis (Nordst) Geitl, CALU-835 Date din literatură 0,36 - - - - Indici estimaţi 0,34…0,42 0,54…0,60 0,45…0,48 0,94…1,05 1,12…1,36 Spirulina platensis (Nordst) Geitl CNM-CB-01 Date din literatură 6,00…6,30 1,00…1,20 - - - Indici estimaţi 4,70…6,40 1,15…1,22 0,60…0,73 1,35…1,62 2,33…3,45 Strain Glutamic acid γ-linolenic acid Phosphatidylinositol Phosphatidyl-choline Sulfated polysaccharides Spirulina platensis (Nordst) Geitl, CALU-835 Literature data 0.36 - - - - Estimated indices 0.34...0.42 0.54...0.60 0 .45…0.48 0.94…1.05 1.12…1.36 Spirulina platensis (Nordst) Geitl CNM-CB-01 Literature data 6.00…6.30 1.00…1.20 - - - Estimated indices 4.70...6.40 1.15...1.22 0.60...0.73 1.35...1.62 2.33...3.45

Problema pe care o rezolvă prezenta invenţie constă în obţinerea unei tulpini noi de Spirulina platensis, biomasa căreia s-ar caracteriza printr-un conţinut înalt de substanţe biologic active, în special de acid glutamic, acid γ-linolenic, fosfatidilinozitol, fosfatidilcolină şi polizaharide sulfatate. The problem that the present invention solves consists in obtaining a new strain of Spirulina platensis, the biomass of which would be characterized by a high content of biologically active substances, especially glutamic acid, γ-linolenic acid, phosphatidylinositol, phosphatidylcholine and sulfated polysaccharides .

Esenţa invenţiei constă în faptul că se propune o tulpină de algă Spirulina platensis (Nordst) Geitl care este depozitată în Colecţia Naţională de Microorganisme Nepatogene a Republicii Moldova a Institutului de Microbiologie şi Biotehnologie al AŞM cu numărul CNMN-CB-11 şi poate fi utilizată în calitate de sursă de acizi glutamic şi γ-linolenic, polizaharide sulfatate, fosfatidilinozitol şi fosfatidilcolină. The essence of the invention consists in the fact that a strain of Spirulina platensis (Nordst) Geitl algae is proposed which is stored in the National Collection of Non-pathogenic Microorganisms of the Republic of Moldova of the Institute of Microbiology and Biotechnology of AŞM with the number CNMN-CB-11 and can be used in as a source of glutamic and γ-linolenic acids, sulfated polysaccharides, phosphatidylinositol and phosphatidylcholine.

Rezultatul constă în sporirea productivităţii algei şi a cantităţii de acizi glutamic şi γ-linolenic, polizaharide sulfatate, fosfatidilinozitol şi fosfatidilcolină. The result consists in increasing the productivity of the algae and the amount of glutamic and γ-linolenic acids, sulfated polysaccharides, phosphatidylinositol and phosphatidylcholine.

Rezultatul tehnic al invenţiei, în comparaţie cu cea mai apropiată soluţie, constă în sporirea conţinutului a cinci componente bioactive de bază în biomasa absolut uscată a cianobacteriei: acid glutamic 8,10…9,64% (faţă de 4,70…6,40% la cea mai apropiată soluţie); acid γ-linolenic 1,36…1,45% (faţă de 1,15…1,22% la cea mai apropiată soluţie); polizaharide sulfatate 4,03…5,01% (faţă de 2,33…3,45% la cea mai apropiată soluţie); fosfatidilinozitol 0,70…0,85% (faţă de 0,60…0,73% la cea mai apropiată soluţie) şi fosfatidilcolină 1,86…2,00% (faţă de 1,35…l,62% la cea mai apropiată soluţie). The technical result of the invention, compared to the closest solution, consists in increasing the content of five basic bioactive components in the absolutely dry biomass of the cyanobacterium: glutamic acid 8.10...9.64% (compared to 4.70...6.40 % to the closest solution); γ-linolenic acid 1.36...1.45% (compared to 1.15...1.22% for the closest solution); sulfated polysaccharides 4.03...5.01% (compared to 2.33...3.45% for the closest solution); phosphatidylinositol 0.70...0.85% (compared to 0.60...0.73% at the closest solution) and phosphatidylcholine 1.86...2.00% (compared to 1.35...l.62% at the closest approximate solution).

Rezultatul tehnic obţinut se datorează particularităţilor fiziologice şi biochimice ale tulpinii, în special proceselor biosintetice, care decurg în acest organism. Ca rezultat al adaptării tulpinii la o temperatură mai joasă de cultivare are loc modificarea ciclurilor biosintetice cu deplasarea lor spre produsele nominalizate. The technical result obtained is due to the physiological and biochemical particularities of the strain, especially the biosynthetic processes that take place in this organism. As a result of the adaptation of the strain to a lower cultivation temperature, the modification of the biosynthetic cycles takes place with their displacement towards the nominated products.

Tulpina propusă a fost colectată dintr-un bazin acvatic artificial din apropierea satului Căpriana. The proposed strain was collected from an artificial water basin near the village of Căpriana.

În cultura algologic pură tulpina se multiplică intens pe mediul mineral lichid cu următoarea componenţă a macroelementelor, g/L: NaNO3 - 2,25; NaHCO3 - 8,0; NaCl - 1,01; K2SO4 - 0,3; NaHPO4 - 0,20; MgSO3·7H2O - 0,20; CaCl2 - 0,024; soluţia de microelemente 1 mL/L, ce conţine în mg/L: H3BO3 - 2,86; MnCl2·4H2O - 1,81; ZnSO4·7H2O - 0,22; CuSO4·5H2O - 0,08; MoO3 - 0,015; Fe-EDTA - l,0 mL/L. In the pure algal culture, the strain multiplies intensively on the liquid mineral medium with the following composition of macroelements, g/L: NaNO3 - 2.25; NaHCO3 - 8.0; NaCl - 1.01; K2SO4 - 0.3; NaHPO4 - 0.20; MgSO3·7H2O - 0.20; CaCl2 - 0.024; microelement solution 1 mL/L, which contains in mg/L: H3BO3 - 2.86; MnCl2·4H2O - 1.81; ZnSO4·7H2O - 0.22; CuSO4·5H2O - 0.08; MoO3 - 0.015; Fe-EDTA - l.0 mL/L.

În condiţii optime de cultivare tulpina respectivă are o productivitate de 1,5…1,6 g/L cu conţinut de acid glutamic 8,10…9,64%; acid γ-linolenic 1,36…1,45%; polizaharide sulfatate 4,03…5,01%, fosfatidilinozitol 0,70…0,85%, şi fosfatidilcolină 1,86…2,00%. In optimal cultivation conditions, the respective strain has a productivity of 1.5...1.6 g/L with a glutamic acid content of 8.10...9.64%; γ-linolenic acid 1.36...1.45%; sulfated polysaccharides 4.03...5.01%, phosphatidylinositol 0.70...0.85%, and phosphatidylcholine 1.86...2.00%.

Caracterele morfo - culturale ale tulpinii Morpho-cultural characters of the strain

Tulpina prezintă forme filamentoase homocite de culoare verde închisă. Trihomele sunt alcătuite din celule cilindrice (lungimea 8…10 µm, lăţimea 6…8 µm) absolut identice, cu excepţia celulelor terminale, care se îngustează spre capătul trihomului şi sunt uşor rotunjite. Creşterea lor are loc ca rezultat al diviziunii transversale a celulelor. Celulele conţin granule, situate în apropierea pereţilor celulari. The stem shows dark green homocytic filamentous forms. The trichomes are made up of cylindrical cells (length 8...10 µm, width 6...8 µm) absolutely identical, except for the terminal cells, which narrow towards the end of the trichome and are slightly rounded. Their growth occurs as a result of the transverse division of cells. The cells contain granules, located near the cell walls.

Trihomele sunt separate, nu se ramifică şi nu se răsucesc în formă de spirală. Diametrul trihomei este de 6…8 µm, iar lungimea de 0,14 …1,30 mm. Ele nu formează teci mucozitare. The trichomes are separate, do not branch and do not twist in a spiral shape. The diameter of the trichome is 6...8 µm, and the length is 0.14...1.30 mm. They do not form mucocytic sheaths.

În mediu lichid formează suspensie practic uniformă cu tendinţă neînsemnată de acumulare la hotarul lichid-aer. In a liquid environment, it forms a practically uniform suspension with an insignificant tendency to accumulate at the liquid-air boundary.

Caracterele fiziologo - biochimice Physiological - biochemical characters

Tulpina microalgei verzi Spirulina platensis (Nordst) GEITL CNMN-CB-11 creşte bine pe mediul SP1, componenţa căruia a fost menţionată mai sus. The green microalga strain Spirulina platensis (Nordst) GEITL CNMN-CB-11 grows well on the SP1 medium, the composition of which was mentioned above.

Temperatura optimă este de 26±2°C, pH 8…9. Intensitatea optimă a luminii este de 1500…2000 lx. The optimal temperature is 26±2°C, pH 8...9. The optimal light intensity is 1500...2000 lx.

Tulpina se adaptează uşor la variaţii neînsemnate ale condiţiilor de cultivare. The strain easily adapts to insignificant variations in cultivation conditions.

În condiţii optime de cultivare tulpina respectivă are o productivitate de 1,5…1,6 g/L cu conţinut de acid glutamic 8,10…9,64%; acid γ-linolenic 1,36…1,45%; polizaharide sulfatate 4,03…5,01%, fosfatidilinozitol 0,70…0,85% şi fosfatidilcolină 1,86…2,00%. In optimal cultivation conditions, the respective strain has a productivity of 1.5...1.6 g/L with a glutamic acid content of 8.10...9.64%; γ-linolenic acid 1.36...1.45%; sulfated polysaccharides 4.03...5.01%, phosphatidylinositol 0.70...0.85% and phosphatidylcholine 1.86...2.00%.

Gradul de puritate a tulpinii The degree of purity of the strain

Tulpina a fost obţinută prin metoda "pasaje repetate" în cultura algologic pură şi acumulată în mediul lichid şi agarizat SP1. La cultivarea în masă poate fi infectată cu alge şi cianobacterii. Contaminarea poate fi evitată prin respectarea condiţiilor de alcalinitate a mediului de cultură. The strain was obtained by the "repeated passages" method in pure algal culture and accumulated in SP1 liquid and agar medium. In mass cultivation it can be infected with algae and cyanobacteria. Contamination can be avoided by respecting the alkalinity conditions of the culture medium.

Exemplu de realizare a invenţiei Example of realization of the invention

În vasele de cultură cu un volum de 1 L cu mediu mineral lichid al macro- şi microelementelor: macroelemente cu următoarea componenţă, g/L: NaNO3 - 2,25; NaHCO3 - 8,0; NaCl - 1,01; K2SO4 - 0,3; NaHPO4 - 0,20; MgSO3·7H2O - 0,20; CaCl2 - 0,024; microelemente - 1 mL/L, cu următoarea componenţă, mg/L: Н3ВО3 - 2,86; MnCl2·4H2O - 1,81; ZnSO4·7H2O - 0,22; CuSO4·5H2O - 0,08; MoO3 - 0,015; Fe-EDTA - l,0 mL/L, pregătit pe apă distilată, se introduce inoculum de 0,40…0,45 g/L în recalcul la biomasa absolut uscată. Cultivarea se realizează la intensitatea luminii de 1500…2000 lx timp de 6 zile, la temperatura de 26±1°C, în ziua a 6-ea productivitatea ajunge până la 1,6 g/L biomasă absolut uscată, cu conţinut de acid glutamic 9,3%; acid γ-linolenic 1,39%; polizaharide sulfatate 4,76%, fosfatidilinozitol 0,79% şi fosfatidilcolină 1,94%. In culture vessels with a volume of 1 L with liquid mineral medium of macro- and microelements: macroelements with the following composition, g/L: NaNO3 - 2.25; NaHCO3 - 8.0; NaCl - 1.01; K2SO4 - 0.3; NaHPO4 - 0.20; MgSO3·7H2O - 0.20; CaCl2 - 0.024; trace elements - 1 mL/L, with the following composition, mg/L: Н3ВО3 - 2.86; MnCl2·4H2O - 1.81; ZnSO4·7H2O - 0.22; CuSO4·5H2O - 0.08; MoO3 - 0.015; Fe-EDTA - 1.0 mL/L, prepared on distilled water, introduce inoculum of 0.40...0.45 g/L in recalculation to absolutely dry biomass. Cultivation is carried out at a light intensity of 1500...2000 lx for 6 days, at a temperature of 26±1°C, on the 6th day the productivity reaches up to 1.6 g/L absolutely dry biomass, with glutamic acid content 9.3%; γ-linolenic acid 1.39%; sulfated polysaccharides 4.76%, phosphatidylinositol 0.79% and phosphatidylcholine 1.94%.

1. MD 169 C2 1995.03.31 1. MD 169 C2 1995.03.31

2. Rudic V., Rudi L., Cepoi L. Aspecte biotehnologice de reglare a biosintezei acizilor graşi polienici la Spirulina platensis CNM-CB-01. Buletinul Academiei de Ştiinţe a Moldovei. Seria Ştiinţele vieţii. Chişinău, 2007, nr.1 (301), p. 135-142 2. Rudic V., Rudi L., Cepoi L. Biotechnological aspects of regulating the biosynthesis of polyenic fatty acids in Spirulina platensis CNM-CB-01. Bulletin of the Academy of Sciences of Moldova. Life Sciences Series. Chisinau, 2007, no. 1 (301), p. 135-142

Claims (1)

Tulpină de algă Spirulina platensis (Nordst) Geitl CNMN-CB-11 în calitate de sursă de substanţe biologic active.Algae strain Spirulina platensis (Nordst) Geitl CNMN-CB-11 as a source of biologically active substances.
MDA20110016A 2011-02-24 2011-02-24 Strain of Spirulina platensis (Nordst) Geitl alga as a source of biologically active substances MD4122C1 (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
MD4334C1 (en) * 2014-02-03 2015-10-31 Государственный Университет Молд0 Strain of blue-green microalga Cylindrospermum licheniforme (Bory) Kütz - source of glucides
MD4351C1 (en) * 2014-04-08 2016-01-31 Государственный Университет Молд0 Strain of blue-green microalga Calothrix elenkinii Kossinsk. - source of glucides

Families Citing this family (1)

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Publication number Priority date Publication date Assignee Title
MD4319C1 (en) * 2013-10-17 2015-08-31 Государственный Университет Молд0 Bis[(iminodiacetate)oxothiomolybdate(V)]tetraphenylphosphonium hydrate and process for cultivation of Spirulina platensis cyanobacterium with the use thereof

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MD169C2 (en) * 1985-04-10 1995-07-31 Государственный Университет Молд0 The stem of alga Spirulina platensis (Nordst) Geitl CALU - biomass producer

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MD169C2 (en) * 1985-04-10 1995-07-31 Государственный Университет Молд0 The stem of alga Spirulina platensis (Nordst) Geitl CALU - biomass producer

Non-Patent Citations (1)

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Title
Rudic V., Rudi L., Cepoi L. Aspecte biotehnologice de reglare a biosintezei acizilor graşi polienici la Spirulina platensis CNM-CB-01. Buletinul Academiei de Ştiinţe a Moldovei. Seria Ştiinţele vieţii. Chişinău, 2007, nr.1 (301), p. 135-142 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
MD4334C1 (en) * 2014-02-03 2015-10-31 Государственный Университет Молд0 Strain of blue-green microalga Cylindrospermum licheniforme (Bory) Kütz - source of glucides
MD4351C1 (en) * 2014-04-08 2016-01-31 Государственный Университет Молд0 Strain of blue-green microalga Calothrix elenkinii Kossinsk. - source of glucides

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Owner name: "VIORICA-COSMETIC" S.A., MD STR. MESAGER NR.1, MD-

Free format text: PREVIOUS PROPRIETOR: RUDIC VALERIU, MD