EA017602B1 - Arthrospira maxima buta strain for production of biomass - Google Patents

Abstract

The invention relates to biotechnology, in particular to the production of biomass for use in different spheres of human activity, for example in medicine, cosmetology, sports, livestock farming, bee-keeping, fish breeding, poultry, veterinary etc. A thermophilic strain of CCAP 1406/3 Arthrospira maxima Buta cyanobacteria is produced from a natural relict water reservoir at Apsheron peninsula of the Azerbaijan Republic. A method for producing blue-green microalga biomass Arthrospira maxima Buta by photosynthesis in aqueous nutrient medium is characterized in growing, mixing of microalga suspension, periodic feeding of the medium by the nutrient solution and biomass takeoff after which the aqueous nutrient medium is reclaimed by reversed osmosis.

Description

(57) The invention relates to biotechnology, to the production of biomass with a wide range of uses in various fields of human activity: medicine, cosmetics, sports, animal husbandry, beekeeping, fish farming, poultry farming, veterinary medicine, etc. The thermophilic strain of cyanobacteria Aygokrya mine Vi1a, SSAR 1406/3 , bred in a natural relict reservoir on the Absheron peninsula of the Republic of Azerbaijan. The method of producing biomass of the blue-green microalgae Aygokrya Vi1a mine by photosynthesis in an aqueous nutrient medium consists in growing, mixing a suspension of microalgae, periodically feeding the medium with a nutrient solution and selecting biomass, after which the aqueous nutrient medium is subjected to reclamation by reverse osmosis.

017 602 Β1

017 602 B1

The invention relates to biotechnology, specifically to the cyanobacteria Lt1to8r1ga takh1ta, and is intended for the production and processing of biomass as an additive in feed for poultry, livestock, fish farming and sericulture. The biomass of AgStokrya tah1ta [4, 9, 11, 12, 13] (as a finished product for consumption) is used in various fields of human activity: medicine, cosmetics, sports, animal husbandry, beekeeping, fish farming, poultry farming, veterinary medicine, etc.

A known strain of blue-green algae AtITocrt tachlata (Suaiabispa, OxShaFpakk) 8c (cbc11 c1 Sagbieg 81gat (strain) COMRECE 1968/3786 isolated from Lake Bod (C1ab, 1aque Bob) in Chad (Africa) ) [pkShShs o! Vo! aiu, Asabstu o! 8s1spss5 o! 11s SkhesN KsriBs, Sow o! Р1усо1ода, ЭikeНк, 135, C2-379 82, and 8ры1ш ркИсщий (Sot.) Сс11 (deposited at the Institute of Physiology Institute Russian Federation under registration number 1РРАЗ В-437), whose biomass is used as an additive to animal and poultry feeds. This strain possesses typical morphological, physiological and biochemical characteristics inherent in the blue-green algae of the genus Zryi Nia.The relative disadvantage of this strain is the relatively low level of biomass accumulation under industrial cultivation of 0.3-0.5 g (abs. Dry matter) / l per day and in instability in conditions of a long continuous culture of pronounced polymorphism, a significant decrease in productivity.

The objective of the invention is to obtain a thermophilic strain of cyanobacteria AiItokrt takh1ta, with increased ability to accumulate biomass and ensuring its large-scale industrial cultivation on a non-waste basis.

The task is achieved by the thermophilic strain of cyanobacteria AgStokrya takh1ta, the author's name Vya, bred in the conditions of the natural relict lake Boyuk Shor, Absheron peninsula (Azerbaijan Republic). Atstokrt ottoman Vi! A is a filamentous planktonic cyanobacterium (Suaiioispa). The strain AtShtokri-a ottoman Vi1a is deposited in CiIs SoPssyop about! A1das apb PrgoYuhoa 8AM8 Keksatsy Zeguyusk Yb. Zsoiin Akkoaaboi! Og Mapis 8s1ssiss Zsoiyin Mapis 1p5Si1s EipSd, Agdu11, PA37 1CA, uk \ Ush glad: \ y \ y \ u.5at5.as.ik / Sa1a1odis \ uh uh: 1Wp: // \ y \ y \ y. ssar.as.ik TS1: (44) 1631 559000, Epesus! b1a1: (44) 1631 559386, Eax: (44) 1631 559001 under the SSAR number 1406/3.

Morphological signs. Apitokrya takhta Vya, like all prokaryotes, has a low level of cellular differentiation (there are no chromatophores, a true nucleus, nucleoli, vacuoles, mitochondria, endoplasmic reticulum, etc.). Non-branching spiral-shaped trichomes (filaments, or filaments) of cylindrical cells are surrounded by a mucous membrane and are capable of sliding and rotational movement. When exposed to various physical and chemical factors, filaments can straighten out. Typical pigments, in addition to chlorophyll and carotenoids, are phycobiliproteins. The sexual process in cyanobacteria is absent. Apytokrya otta Vyya propagates with the help of hormones of short-chain, capable of movement, sections of threads formed by fragmentation of maternal trichomes into necridia (specialized cells undergoing lysis).

Physiological and biochemical properties Apigokrya ottoman Vi! A. Nitrogen does not fix; uses nitrate and ammonium nitrogen as a source of nitrogen nutrition. It uses bicarbonates (mainly), carbonates, organic compounds (even glucose) as carbon sources. Maximum biomass growth occurs in sunlight. It also grows when using light sources of any spectral composition. Optimum growth when using a light source close in spectral composition to the sun. The photoperiod is not expressed, and there is no seasonality. With the accumulation of biomass, the culture of the Atshtokrpa cells of Ottoman Vya floats (floats), which is an advantage for the industrial production of biomass of this strain. It grows in the salinity range from 0 to 0.2 M (No. C1). Optimum growth at 0.02 M (NaCl). There is an increase in the range at pH from 6.0 to 12.0; optimum at pH 9.5-10.5. Growth is observed in the temperature range from 20 to 45 ° C; optimum 30-35 ° C. Inhibition and death of the culture occurs at temperatures above 50 ° C.

The chemical composition of AgStokrya ottoman Vya cells depends on the growing conditions and the composition of the nutrient medium [5]. Parametric control in biomass production gives a wide range of composition (in%):

protein - 50.0-75.0, carbohydrates - 12.0-20.0, fats (lipids) - 5.0-7.0, nucleic acids - 5.5-6.5, chlorophyll (a) - 0 , 9-0.95, ash - 7.0.

Component g / 100:

nitrogen - 8-10;

phosphorus - 1-1.1; sodium - 1-2; potassium - 1-2;

magnesium - 0.2-0.3;

- 1 017602 calcium - 0.8-1.0;

sulfur - 0.5-0.6.

Component mg / kg:

iron - 800-1900;

copper - 11-15;

Manganese - 44-60;

nickel - 3-5; zinc - 22-30; cobalt - 1-1.5; chlorine - 2-5; chrome - 3-5; iodine - 500-100.

Storage conditions.

The strain is stored on agarized nutrient media and in a depleted nutrient medium ΟΝ (g / l: sodium nitrate - 3.0; potassium hydrogen phosphate - 0.2; magnesium sulfate - 0.5; sodium bicarbonate - 16.0; calcium chloride - 0, 02; sodium chloride - 1.0; ferrous sulfate (11) - 0.01; sodium salt of ethylenediaminetetraacetate (EDTA) - 0.03; boric acid - 0.0029; manganese chloride - 0.0018; zinc sulfate - 0.0002 ; ammonium molybdenum acid - 0.00003, cobalt chloride - 0.00005).

The strain Atshtovrta takh1ta Vi1a has a wide range of cultivation temperature (20-45 ° C). There is no seasonality in its reproduction. The strain is not picky about the nutrient medium. The strain has excellent flotation (cells float) - biomass is easy to collect and process.

A method for producing biomass of Aiitovrpa tachlata Vi1a includes the cultivation (photosynthesis) of microalgae biomass in a nutrient solution of inorganic salts, providing lighting and thermostating (heating) mode of the cultured nutrient medium to obtain a given composition; design and construction features of bioreactors; design decisions, the composition of the cultivated medium; drying and packaging. The main structure for biomass production - the bioreactor - is a concrete tank with a liquid depth of 0.55 m, a width of 4 m and a length of 10 m. The capacity of this basin bioreactor is from 16 to 18 m ^{3 of} suspension. The pool is divided by a longitudinal partition 0.5 m high, 0.2 m thick and 8 m long. It has a sliding cover system with a transparent polymer film (arcuate design, if necessary, can be automated). Mixing the suspension of microalgae is carried out mechanically and / or by sparging with air. Thermostating (heating) of the suspension (cultured fluid) is ensured by solar lighting, as well as a backup electric heating system. The constructions are reservoirs, pumps, communications, modules for the cultivation of salts, a block of uterine culture (pure seed). A method for producing biomass involves the absorption of carbon dioxide from air. The resulting biomass is separated from the cultivated medium by separation through gauze trays or using a flow centrifuge. Biomass is dried on special pallets or in a spray dryer. On average, the method for producing biomass of microalgae (cyanobacterium) AtStovrpa tachlata Vi1a for use in the food industry, pharmacology and agriculture is 8-10 days.

The method of producing biomass AITovrpa takh1ta Vi1a in experimental industrial conditions is presented by the following examples of its implementation.

Example 1. The strain of blue-green microalgae AtShtovrta takh1ta Vi1a is grown in a glass tube cultivator with a volume of 15 l in a cumulative mode on medium содержащей containing, g / l: sodium nitrate - 3.0; potassium hydrogen phosphate - 0.2; magnesium sulfate - 0.5; sodium bicarbonate - 16.0; calcium chloride 0.02; sodium chloride - 1.0; iron sulfate (11) - 0.01; sodium salt of ethylenediaminetetraacetate (EDTA) 0.03; boric acid - 0.0029; manganese chloride - 0.0018; zinc sulfate - 0,0002; ammonium molybdenum acid - 0.00003, cobalt chloride - 0.00005.

The cultivation was carried out under constant irradiation with T3 8YM 8P1KAE E27 lamps at a temperature of 25-30 ° C and a pH of 9.0. The mixing process by a peristaltic pump. The air exchange rate is 70 g / l of suspension / h (compressor). The saturation of CO ₂ (carbon dioxide) is carried out using an air mixture of air. The total cultivation cycle is 30 days. Biomass selection begins on the 3rd day of cultivation. The average level of biomass accumulation is 1.8-2.0 g asv / l per day.

Example 2. The strain of microalgae A51etotoia5 vayia Myha1ab | grown in a bioreactor. Thermostating (heating) of the suspension (cultured fluid) is ensured by solar lighting, as well as a backup electric heating system. Biomass production is carried out on medium среде in the accumulative mode. Cultivation is carried out at a pH of 9.5, a temperature of 25-30 ° C and natural sunlight (daylight hours 8-10 hours) for 10 days (spring and autumn) and 7-8 days (daylight hours 12 hours) in the summer. The temperature regime day-night is about 5 ° C. At the end of the process, the biomass yield was 80 kg (spring and autumn) - 100 kg (summer).

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The data presented indicate significant physiological and morphological differences of the proposed thermophilic strain of cyanobacterium LtStokrpa shakh1sha Vi! And from the known and its technological advantages over the known strains of blue-green algae [1]. The wide range of applicability of AgFogocrya tachyta Vi! A consists of two main areas: the use of biomass itself and the use of biomass as raw materials for the production of any valuable substances [7, 8, 9].

Literature.

1. 2atgoisk, S. Soiytbiyoi a 1'e [ibe b'ie suaior. 1pDieise bebyk pHukk | iek e [s1shpk | is5 kit 1a sokkaise e! 1a rko1o8yi1e8e be 8ryyikia takh1ta / S. 2aggoisk // RN.E. Fake. Rapk. 1966.

2. C1ethey! 6. Rtobisyoy aib with Nagas1eg1k (1s soiqfiy oG [Do not give 8ril1ta p1a1eik1k aib tah1ta. Aiia1ek be 1a Yibyuy e [be GANTeShayoi. Wo. 29. IO. 6.O. 1975.

3. Nibkoy V.K. Kapk 1.6. Tne Pr1bk oG [Not a1da 8r1gi1ta. 1. 8c1eise oG robo & adpsiNig. wo1. 25. Io. 7.1974.

4. 8ekNabps S.U. aib Tnak 8. Mack sixt oG 8rj1ta iktd 1o \\ - juice [sixteen. Vu [ecH1y1odu be1 [eg. wo1. 11.1979.

5. Weskeg. Ε.ν. This is the library of OG T1SGOA1DAY. VI. 198 Radek. 1985.

6. Totake S .. booooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooooo A1da1 Vu [eNGyu1odu. 1987.

7. CONNECTIONS Ζ .. VönkNak A. .. KtsNoib A. Raiu aslb sotrokyyui oG 8ryi1sha kyashka dgo ^ and uibeg uapoik eiν ^^ oηteiΐa1 soibyuik. RNUUSetNetMgu. wo1. 26.1987.

8. There is no. .. Th1. O. Th. Linden tree. Reu. M1stoYo1 .. then1. 39.1985.

9. VopkNak A. .. Ktsntoib A. Makk rtobisyoi og Fe L1ie-dgeei a1da 8ryi1sha: ai oueg \ ze \\ ·. Wutack. go1. 15.1988.

10. Fuck ^ A.K. A.Ryu1ta p1a1eik1k (AgFgokrya): RNuk1o1odu. Ce11-b1o1odu aib vu [esnguyodu. Loboy: Bk1o1. Tau1og & Rgais1k. 233 Radek. 1997.

11. MoogNeab K.1. Mogdai N.S. 8 и ша 1 ша и ге ге ге ки ки ки рег рег Г RNKNeb uy Shyekh. 1 fig. I8A. 1995.

12. Vojoe LJ .. Sak [eiNo1 / Ρ. \ Ν .. Vegdeu'k Mayia1 oG 8uk1etays Vas1egu1odu. go1. 1. ΝΥ. Unbehind. Branches: 8rttdeg ^ er1ad. 2001.

13. 6egkN \\ 'H1 M.E. (Ituetkyu OG SayGotsha. Ea \ zk. I8A); Be1au Atka (EatFike Miyyuia1k. Sayrayta. SayGotsha. I8A) 8ryi1sha F Nitai Miyyuyi aib Nea1F. Radek 328. 2007.

Claims (1)

CLAIM Strain AgFgokrya ottoman Vi! And SSAP 1406/3 for biomass production. Eurasian Patent Organization, EAPO Russia, 109012, Moscow, Maly Cherkassky per., 2