ISSN 2074-9414 (Print),
ISSN 2313-1748 (Online)

Low–Temperature Storage and the Viability Preservation of Streptomyces

Abstract
The most effective way to store microorganisms of different taxonomic groups is at low temperatures from minus 12°C to minus 150°C. The present research features the influence of low temperature (minus 12°C and 18°C) on the viability of collection strains of actinomycetes Streptomyces lucensis VKPM Ac-1743 and Streptomyces violaceus VKPM Ac-1734, producers of glycosidase inhibitors. The strains were stored without a cryoprotector in a 15% glycerol solution and 0.9% sodium chloride aqueous liquid. The research objective was to check their ability to keep their inhibitor activity against pancreatic amylase during corn starch hydrolysate fermentation. The experiment made it possible to determine the titer (CFU in 1 cm3 of the initial inoculum) and inhibitory activity against pancreatic α-amylase. It was revealed that Streptomyces lucensis and Streptomyces violaceus strains in cell initial concentrations of 107 and 108 CFU/cm3 maintained high viability level during four months conservation in 15% glycerol solution and 0.9% sodium chloride aqueous solution at the temperatures of minus 12 °C and minus 18 °C. Most cells survived at the conservation in a 15% glycerol solution at minus 18 °C. The inhibitor activity level in cultural liquid was higher in Streptomyces lucensis and Streptomyces violaceus strains kept in 15% glycerol solution at low the temperatures than in those kept in a 0.9% sodium chloride solution. The cultures kept in a 15% glycerol solution at minus 18 °C had higher inhibitor activity indicators 2600 ± 200 IU/cm3 . The research proved that low-temperature storage of Streptomyces produces no negative effect on the viability and biosynthetic activity of the cultures.
Keywords
Strains of Streptomyces lucensis VKPM Ac-1743 and Streptomyces violaceus VKPM Ac-1734, low–temperature storage, viability, inhibitory activity, titer
REFERENCES
  1. Sidyakina T.M. Konservatsiya mikroorganizmov v kollektsiyakh kulʹtur [Preservation of microorganisms in the collectionsof cultures]. Sbornik nauchnykh trudov “Konservatsiya geneticheskikh resursov. Metody. Problemy. Perspektivy” [Collection ofscientific Proceedings “Conservation of genetic resources. Methods. Problems. Prospects”]. Pushchino, 1991, pp. 81–159. (In Russ.).
  2. Sharova N.Yu., Vybornova T.V., Printseva A.A., and Manzhieva B.S. The properties of the conidia of strains of theactinomycete Streptomyces lucensis and Streptomyces violaceus during storage at low temperatures. Food systems, 2018, vol. 1, no. 3,pp. 27–32. (In Russ.). DOI: https://doi.org/10.21323/2618-9771-2018-1-3-27-32.
  3. Filippova S.N., Surgucheva N.A., and Gal’chenko V.F. Long-term storage of collection cultures of actinobacteria.Microbiology, 2012, vol. 81, no. 5, pp. 682–690. (In Russ.).
  4. Tsutsaeva A.A., Anan’ina A.E., Balyberdina L.M., Stepanyuk L.V., and Pavlenko N.V. Long-term storage of industrialmicrobial strains. Microbiology, 2008, vol. 77, no. 5, pp. 696–700. (In Russ.).
  5. Chukparova A.U. Otsenka sokhraneniya zhiznesposobnosti shtammov mikroorganizmov pri nizkotemperaturnoykonservatsii [Evaluation of the viability preservation of microorganism strains during low-temperature conservation]. MaterialyVserossiyskogo simpoziuma s mezhdunarodnym uchastiem “Biologicheski aktivnye veshchestva mikroorganizmov – proshloe,nastoyashchee, budushchee” [Materials of the All-Russian Symposium with international participation [Biologically active substancesof microorganisms – past, present, future”]. Moscow, 2011, p. 131. (In Russ.).
  6. Shenderov B.A., Gakhova Eh.N., Manvelova M.A., Piorunskij D.A., and Karnaukhov V.N. Method of the prolonged storageof natural symbiotic of human and animal microorganism associations. Patent RF, no. 2123044, 1998.
  7. Arkadʹeva Z.A., Bezborodov A.M., Blokhina I.N., et al. Promyshlennaya mikrobiologiya [Industrial Microbiology].Moscow: Higher School Publ., 1989. 688 p. (In Russ.).
  8. Sineva O.N., Kulikova N.G., Filippova S.N., and Terekhova L.P. Storage of Actinobacteria of the Genera Streptomycesand Nonomuraea by Low Temperature Preservation. Antibiotics and Chemotherapy, 2014, vol. 59, no. 11–12, pp. 11–15. (In Russ.).
  9. Sharova N.Yu. Amylase inhibitors from Streptomyces lucensis VKPM Ac-1743 and Streptomyces violaceus VKPM Ac1734. Applied Biochemistry and Microbiology, 2015, vol. 51, no. 1, pp. 58–63. DOI: https://doi.org/10.1134/S0003683815010159.
  10. Sharova N.Yu., Nikiforova T.A., and Pozdnyakova T.A. Shtamm aktinomitseta Streptomyces violaceus — produtsentingibitora glikozidaz [The strain of actinomycete of Streptomyces violaceus, an glycosidase inhibitor producer]. Patent RF,no. 2346042, 2009.
  11. Sharova N.Yu, Pozdnjakova T.A., and Khodkevich O.A. Actinomycete strain of Streptomyces lucensis – producer ofglycosidase inhibitor. Patent RF, no. 2355755, 2009.
  12. State Standart 10444.15-94. Food products. Methods for determination of quantity of mesophilic aerobes and facultativeanaerobes. Moscow: Standartinform Publ., 2010.
  13. Khodkevich O.A. Razrabotka tekhnologii biosinteza ingibitora α-glikozidaz aktinomitsetami roda Streptomyces iprimenenie kompleksnoy dobavki na ego osnove v khlebopechenii. Diss. kand. tekhn. nauk [Development of the biosynthesis technologyof α-glycosidase inhibitor by actinomycetes of the genus Streptomyces and the use of complex additives in bread making: Cand. Tech. Sci. Dis]. St.Petersburg, 2009. 135 p.
  14. Akulova N.Yu. Ingibitory α-glyukozidaz iz Streptomyces. Vydelenie i svoystva. Avtoref. dis. kand. biol. nauk [Inhibitors of α-glucosidase from Streptomyces. Allocation and properties: Cand. Biol. Sci. Diss]. St.Petersburg, 1993. 22 p.
  15. Sharova N.Yu. Razrabotka nauchnykh osnov novykh tekhnologiy pishchevykh dobavok i ingredientov s ispolʹzovaniem krakhmalsoderzhashchego syrʹya. Avtoref. dis. dokt. biol. nauk [Development of the scientific foundations for new technologies of food additives and ingredients with starch-containing raw materials: author. Doct. Biol. Sci. Diss]. St.Petersburg, 2013. 32 p.
  16. Pokhilenko V.D., Baranov A.M., and Detushev K.V. Metody dlitelʹnogo khraneniya kollektsionnykh kulʹtur mikroorganizmov i tendentsii razvitiya [Methods of long-term storage of collection cultures of microorganisms and the development trends]. University proceedings. Volga region. Medical sciences, 2009, vol. 12, no. 4, pp. 99–12. (In Russ.).
  17. Cotarlet M., Bahrim G., Negoita T., and Stougaard P. Screening of polar streptomycetes able to produce cold-active hydrolytic enzymes using common and chromogenic substrates. Romanian Biotechnological Letters, 2008, vol. 13, no. 5, pp. 69–80.
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