EA017600B1 - Thermophilic strain of blue-green algae spirulina platensis baku 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 blue-green microagae CCAP 1475/12 Spirulina platensis Baku is produced by a direct extraction from a natural water reservoir at Apsheron peninsula of the Azerbaijan Republic. A method for producing microalga biomass Spirulina platensis Baku 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

The invention relates to biotechnology, specifically to δρίπιΐίηα p1a1ep818, and is intended for the production and processing of biomass as an additive in feed for poultry, livestock, fish farming and sericulture. Biomass 8rnikpa p1a1ep818 [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 8rn1sha p1a1ep818 (Sot.) 6ey1 (deposited in the Collection of the Institute of Plant Physiology of the Academy of Sciences of the Russian Federation under registration number ΙΡΡΑ8 B-437), the biomass of which is used as an additive to animal and bird feeds. This strain has the typical morphological, physiological and biochemical characteristics inherent in the blue-green algae of the genus 8p1ti1sha. The relative disadvantage of the known strain is the relatively low level of biomass accumulation under conditions of industrial cultivation of 0.3-0.5 g (abs. Dry matter) / liter per day and instability in conditions of long-term continuous culture, pronounced polymorphism, a significant decrease in productivity.

The task is achieved by the thermophilic strain of cyanobacteria 8rnikpa p1a1ep818, the author’s name is Waki, obtained by direct isolation from a natural lake (Lokbatan, Absheron peninsula, Azerbaijan Republic). Strain 8rni1sha p1a1ep818 Vaki was deposited in Sikit Co11ecΐίοη οί A1day apb PrgoYuhoa 8AM8 Kekeagsk Zetuyuk Y6. 8soSh§k Akkoaakop Gog Mappe 8s1epse 8so1Shk Maghe 1p81ki1e Hypod, Agdu11, ΡΑ37 10A, υκ XV e be glad: te ^^. Te1: (44) 1631 559000, I1ec1 Фа1: (44) 1631 559386, Bach: (44) 1631 559001 under SSAR number 1475/12.

Morphological features of the claimed strain.

8p1sh1sha p1a1ep818 Waki, like all prokaryotes, has a low level of cell differentiation (there are no chromatophores, a true nucleus, nucleoli, vacuoles, mitochondria, an 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. 8rnkpa rkNepMz Waki propagates with the help of hormones - short-chain, capable of moving, sections of filaments formed by fragmentation of maternal trichomes into necridia (specialized cells undergoing lysis).

Physiological and biochemical properties of the 8th r.

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 that is close in spectral composition to the sun. The photoperiod is not expressed and there is no seasonality. Due to the pronounced gas vacuoles, the cell culture (biomass) floats (floats). Which is an advantage for the industrial production of biomass of this strain. It grows in the salinity range from 0 to 0.5 M (IaS1). The optimum growth at 0.02 M (IaS1). 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 occur at temperatures above 55 ° C.

The chemical composition of 8p1gikpa p1a1ep818 Vaki 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 percent):

protein 55.0-75.0;

carbohydrates 10.0-20.0;

fats (lipids) 5.0-7.0;

nucleic acids 5.5-6.5;

chlorophyll (a) 0.9-0.95;

ash content 7.0;

humidity 6.0-8.0;

including fat-soluble pigments 2.0-2.5;

beta-carotene 0.5-0.8; water soluble pigments 3.0-6.0;

ash 7.0-8.0.

Cells also contain:

vitamins of the cyancobalamin group 2 g / g asv;

ascorbic acid 80 mg / 100 g asv;

component g / 100;

- 1 017600 nitrogen 8-10;

phosphorus 1-1.1; sodium 1-2;

potassium 1-2;

magnesium 0.2-0.3;

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 0.6; magnesium sulfate 0.6; sodium hydrogen carbonate 16.0; calcium chloride 0.04; sodium chloride 3 , 0; iron sulfate (11) 0.01; sodium salt of ethylene diamine tetraacetate (EDTA) 0.03; boric acid 0.0029; manganese chloride 0.0018; zinc sulfate 0.0002; ammonium molybdenum acid 0.00003, cobalt chloride 0.00005).

A method of producing spirulina biomass (actually aquaculture) includes growing (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 pool is divided by a longitudinal partition 0.5 m high, 0.2 m thick and 8 m long. It has a system sliding cover 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 by 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) Zriya sha ρΐαίοηκίκ Waki for use in the food industry, pharmacology and agriculture is 8-10 days.

The method of producing biomass Sr1ti1ta rkyspU Vaki in experimental industrial conditions is presented by the following examples of its execution.

Example 1

The strain of blue-green microalgae Zr1ti1ta p1a1sp515 Vaki is grown in a glass tube cultivator with a volume of 15 l in a cumulative mode on nutrient medium ΟΝ containing, g / l: sodium nitrate 3.0; potassium hydrogen phosphate 0.6; magnesium sulfate 0.6; sodium bicarbonate 16.0; calcium chloride 0.04; sodium chloride 3.0; 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.

Growth was carried out under constant irradiation with T3 ZYM ZRGKAY E27 lamps at a temperature of 25-30 ° C and a pH of 9.5. The mixing process is carried out periodically by microcompressors. The air exchange rate of 70 g / l suspension / h Saturation with air and / or CO _{2 is} carried out by bubbling. 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. The general cultivation cycle is 30 days, after which recultivation of the culture medium with reverse osmosis is carried out.

Example 2. The strain of blue-green microalgae Zriyta rkIsschsk Waki grown in a bioreactor. Obtaining biomass is carried out in a cumulative mode on a nutrient medium ΟΝ.

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. Temperature day-night with

- 2017600 sets about 5 ° C. At the end of the process, the biomass yield is 80 kg (spring and autumn) - 100 kg (summer). Then, the culture medium is reclaimed by reverse osmosis.

The data presented indicate significant physiological and morphological differences of the proposed thermophilic strain of cyanobacteria 8ri11pa p1a1epv1v Waki from the known and its technological advantages over the known strains of blue-green algae [1]. The wide range of applicability of the Riikpa p1a1epv1v Waki consists of two main areas: the use of biomass itself and the use of spirulina biomass as raw materials for the production of any valuable substances [7, 8, 9]. The first area includes various methods of using spirulina biomass as a food supplement in the diet of humans and animals [8, 10, 12], using spirulina biomass in medical and biological procedures of a therapeutic and prophylactic nature.

Literature.

1. 2atogoisk S. Soplpi! Yup a 1'e! Ibe b'ipe suaporkusey. 1pDiepse no betkv rkuvshchiev e! skppschiev vig 1a sgovvapse e! 1a rkoYuvuShkeve be 8riikpa takh1ta / S. 2 atgoisk // Rk. Ό. 1kev1v. Rapv, 1966.

2. C1etep! 6. The Archbishop of St. Petersburg, Russia! ! ke a1dae 8ri1sha p1a1epv1v apb takhtaa. App1ev be 1a Maykop e! be 1'Aktep1akop, 1975, yo1. 29, p. 6.

3. Nibwop Wb., Cape 1.6. Tke Nr1bk about! ! ke a1da 8rpi1sha. - 1. 8s1epse oh! Robo & Adpsikige, 1974, yo1. 25, by. 7.

4. 8evkabps S.U. apb Tkotav 8. Mavv sikite oh! 8Riikpa iwshd 1o \\ - sov1 piktikSv, Vyu! Eskpo1 Be1! Egv, 1979, yo1. eleven.

5. Weskeg E. Archbishop apb ive! t1stoa1dae. 1985. VI, 198 Radev.

6. Totavash b., Bauappe b., 8assk1 A., Voss1 E. EGGes1v about! (Ethereum Shge op dgoMk apb скusket1sa1 sotrovyyup ίπ 8riyusha p1a1epv1v vkash M2. A1da1 Vyu eskp1odu, 1987.

7. Sokep Ζ., ^ Pvkak A., Vyuktopb A. Raku as1b sotrovyyup o! 8ppi1sha in! Ga1pv dgo \ up ipbay uapoiv epu1goptep1a1 sopbkupyupv. Rku1osket1vku, 1987, yo1. 26.

8. Sayet O., Tlosh O. Tke юyusket1vku apb tbivktta1 ro! Epk1 o! 8r1ti1ta. - App. Reu. Mutobuck, 1985, wo1. 39.

9. ^ pvkak A., Vyuktopb A. Mavv rgobiskop about! ! ke l1ie-dzheep a1da 8riikpa: ap oye1Me \\ ·. Vyutavv, 1988, yo1. fifteen.

10. Fuck ^ A.A. 8p1gikpa p1a1epv1v (Aiktovrka): Rkuv1odu, Ce11-b1odu apb Vu! eskpo1od, bopbop: W1v! o1, Tau1og & Egaps1v, 1997. - 233 p.

11. Moogkeab KL., Mogdap N.S. 8ii1sha pa! Ige'v viregTob. - Rybkvkeb bk No. kekh, 1ps. I8A, 1995.

12. Voope L.V., Sav1epko1kh V. ^., Vegdeu’s Mapia1 o! 8th! This is you! Heyu1odu, \ ό1. 1. ΝΥ, Heilbäger, Wetkp: 8prtdeg ^ er1ad, 2001.

13. 6egvk \ tt M.E. (Ishuegvyu o! Ca1 ^ Go ^ η ^ a, Jau1b, I8A); Ве1ау Атка (ЕаЛкпве №1попота1в, Sakrakga, Sak! Oshta, I8A) 8riikpa sh Nitap Maikop apb Neakk, 2007, Мпкег о! Radev: 328.

Claims (1)

CLAIM Thermophilic strain of blue-green microalgae 8ri1sha p1a1epv1v Waki SSAR 1475/12 to obtain biomass.