Mikrobiol. Z. 2020; 82(5):3-10.
Marine Actinobacteria – Producers of Enzymes with α-L-Rhamnosidase
L.D. Varbanets1, O.V. Gudzenko1, V.A. Ivanytsia2
1Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine
2Mechnikov Odessa National University
2 Dvoryanskaya Str., Odesa, 65029, Ukraine
In recent years researchers have attracted their attention to such glycosidases as α-L-rhamnosidase (α-L-rhamnoside-rhamnohydrolase – EC 18.104.22.168). The substrates of their action are widespread in the plant world glycosides such as naringin, quercetrin, hesperidin, neohesperidin, and rutin, from which α-L-rhamnosidases cleave the terminal unreduced L-rhamnose residues. This specificity of α-L-rhamnosidases can be used in various industries: food – to improve the quality of drinks (reducing bitterness in citrus juices, enhancing the aroma of wines), as well as production of food additives; in the pharmaceutical industry – to improve the biological properties of bioflavonoids, in particular anti-inflammatory. A number of them are characterized by cardio- and radioprotective effects, have antioxidant, cytotoxic, antibacterial, antisclerotic properties, and are used in the complex treatment of coronary heart disease, including angina pectoris. The use of α-L-rhamnosidases in the chemical industry is associated with a reduction in the cost of rhamnose production as well as various plant glycosides and rutinosides. In the literature available to us, no data were found on the producers of α-L-rhamnosidases among the representatives of actinobacteria, which are known to synthesize a wide range of biologically active compounds, including antibiotics and enzymes. Purpose. To study the ability of actinobacteria isolated from water and bottom sediments of the Black Sea, to produce a-L-rhamnosidase, and also to study the properties of the most active producer. Methods. Glycosidase activity was determined by the Romero and Davis methods, protein – by the Lowry method. Results. The study of 12 glycosidase activities in 10 strains of actinobacteria isolated from bottom sediments of the Black Sea indicated that 6 investigated strains showed the ability to synthesize an enzyme with a-L-rhamnosidase and b-D-glucosidase activity. Studies have shown that the highest α-L-rhamnosidase activity (0.14 U/mg protein) was manifested by Acty 5 isolate with an optimum pH of 7.0 and a temperature optimum of 38°C. The enzyme preparation showed substrate specificity both for natural (rutin, naringin, neohesperidin) and synthetic (p-nitrophenyl derivatives of L-rhamnose and D-glucose) substrates. Conclusions. Promising Acty 5 isolate with high a-L-rhamnosidase and low b-Dglucosidase activity was found among marine actinobacteria. Bacteria with two enzymes activity expand the possibilities of their practical use.
Keywords: actinobacteria, Black Sea, a-L-rhamnosidase.
Full text (PDF, in English)
- Gudzenko OV, Varbanets LD. Microbial α-Lrhamnosidases: producers, properties, practical usage. Biotechnologia. 2012; 5(6):9–26.
- Manzanares P., Valles S., Ramon D., Orejas M. α-L-rhamnosidase: old and new insights. Industrial Enzymes: Springer; 2007. p. 117–140. https://doi.org/10.1007/1-4020-5377-0_8
- Puri M. Updates on naringinase: structural and biotechnological aspects. Appl Microb and Biotechnol. 2012; 93(1):49–60. https://doi.org/10.1007/s00253-011-3679-3
- Yadav V, Yadav PK, Yadav S, Yadav KDS. α-L-Rhamnosidase: A review. Process Biochemistry.2010; 45(8):1226–1235. https://doi.org/10.1016/j.procbio.2010.05.025
- Varbanets LD, Borzova NV. [Glycosidases of microorganisms and research methods]. Kyiv: Nauk. Dumka; 2010 . Ukrainian.
- Borzova NV, Gudzenko OV, Varbanets LD, Nakonechnaya LT, Tugay TI. [Glycosidase and proteolytic activity of micromycetes isolated from the chernobyl exclusion zone]. Microbiol Z. 2020; 82(2):10–15. Ukrainian. https://doi.org/10.15407/microbiolj82.02.051
- Gudzenko OV, Borzova NV, Varbanets LD, Ivanitsia VO, Seifullina II, Pirozhok OV, Chebanenko EA. Glycosidase activity of bacteria and genus Bacillus, isolated from the Black sea. Microbiol Z. 2019; 81(3):14–26. https://doi.org/10.15407/microbiolj81.03.014
- Yadav V, Yadav KDS. New fungal for α-Lrhamnosidase animportant enzyme use dinthesynthesis of drugsand drug precursors. Appl Biochemistry and Microbiol. 2010; 48(3):295–301. https://doi.org/10.1134/S0003683812030155
- Chaplin ME, Kennedy JE. Carbohydrate analysis. Oxford: IRL Press, 1986.
- Davis DW. Determination of flavonones in citrus juice. Anal Biochemistry. 1947; 19(1):46–48. https://doi.org/10.1021/ac60007a016
- Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with folinphenol reagent. J Biol Chem. 1951; 193(2):265–275.
- Lakyn HF. [Byometryia]. Moscow: Vysshaya Shkola; 1990. Russian.