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CHISELIŢĂ, Oleg; BÎRSA, Maxim; VASILICIUC, Anastasia; BURŢEVA, Svetlana. Lioprotetcive potential of cyanobacterial extracts. In: Microbial Biotechnology. Ediția 3, 12-13 octombrie 2016, Chișinău. Chișinău, Republica Moldova: Institutul de Microbiologie şi Biotehnologie, 2016, p. 180.
Ediția 3, 2016
Conferința "Microbial Biotechnology" |
Chișinău, Moldova, 12-13 octombrie 2016
The gene pool of industrial microorganisms is an important natural resource for Republic of Moldova, a heritage of the country. The National Collection of Non-pathogenic Microorganisms (NCNM), as all collections in the world, focuses on the problems of long-term storing and preserving genetic stability of microorganisms that present scientific and practical interest. Choosing appropriate methods of storage of each group of microorganisms is one of the main tasks of collection. Lyophilization and cryopreservation are currently the preferred longterm preservation methods for the most strains of microorganisms. According to experts in the field, reduced viability of microorganisms during storage can be caused by the formation of free radicals. Some authors consider that their occurrence is determined both by changing the ratio of the different forms of water in the cell, and the destruction of the polypeptide chains. Others point that this process occurs due to oxidation reactions. Thus, both free radicals and oxidation products of lipids, by interacting with the cellular components, lead to lesions, causing cell death. To avoid the destruction of the cell during lyophilization, it is necessary to use protective media, which usually contain mineral salts, proteins and / or other substances as polysaccharides, antioxidants, etc. The aim of this research was to assess the lyoprotective potential of various extracts of Spirulina platensis, such as amino acids, oligopeptides and polysaccharides, which possesses antioxidant, anti-radical, cell membranes stabilizing and tissue regenerating actions. There were used two streptomycetes strains stored at NCNM – Streptomyces canosus CNMNAc- 02 and Streptomyces massasporeus CNMN-Ac-06. As a lyoprotectant were tested the extracts of amino acids and oligopeptides (5 mg / ml) and sulfated polysaccharides (10 mg / ml) of Spirulina platensis (Nordst) Getil CNMN-CB-02, which has been added to the standard protective medium – gelatin 2.5% + 7.5% glucose (control medium) in various concentrations (% / V). The viability of Streptomyces massasporeus CNMN-AC-06 after lyophilization increases from 18.4 to 29.0%, compared to the control sample, when used amino acids and oligopeptides extract in a concentration of 0.5; 1 and 2% / V, and from 30.0 to 36.9% – when used sulfated polysaccharides extract in a concentration of 10; 30 to 50% / V. Promising results were obtained in the case of Streptomyces canosus CNMN-Ac-02, as well. The supplementation of protection media with amino acids and oligopeptides extract in a concentration of 0.1; 1 and 10% / V leads to the increase of the viability after lyophilization from 15.3 to 20.1% compared to the control, and from 12.6 to 38.0% when used sulfated polysaccharide in concentration of 5; 10; 20; 30 and 50% / V. Moreover, this effect was shown to be stable over time. Thus, after 1 year of storage using control medium, the viability of studied strains decreased significantly registering 79.1% of the initial number of cells. While cell viability increased by 20,6% -28.3% in the experimental samples using amino acid and peptide extracts and by 20,5% – 39,8% – using sulfated polysaccharides extract. Thus, we can conclude that the studied Spirulina platensis extracts possess a high lyoprotectant potential and can be used to develop protective media for lyophilization of microorganisms. The results of this research will serve to streamline and optimize the methods of storing and preservation of microbial gene pool of NCNM.
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