INFLUENCE OF CATIONS ON NADP+-DEPENDENT GLUTAMATE DEHYDROGENASE ACTIVITY IN BACTERIA OF GENERA ACINETOBACTER, RHODOCOCCUS AND NOCARDIA −PRODUCERS OF SURFACTANTS

Surfactants of microbial origin are widely used in different industries. The application of microbial surfactants is promising in biology and medicine as an alternative to synthetic disinfectants or drugs due to their antimicrobial and antiadhesive properties. The key enzyme biosynthesis of surface active aminolipids (effective antimicrobials preperations) in Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Ac-5017 and Nocardia vaccinii IMV B-7405 is NADP+-dependent glutamate dehydrogenase.

Aim – to study the effect of mono- and divalent cations on the activity of NADP+-dependent glutamate dehydrogenase in Acinetobacter calcoaceticus IMV B-7241, Rhodococcus erythropolis IMV Ac-5017and Nocardia vaccinii IMV B-7405.

The activity of NADP+ -dependent glutamate dehydrogenase (EC 1.4.1.4) in the cell-free extract was analyzed for the formation of glutamate in the oxidation of NADPH at 340 nm.

It was found that activators of this enzyme in A. calcoaceticus ІMV B-7241 were calcium cations (5 mM), magnesium (10 mM) and zinc (0.001–0.01mM), R. erythropolis ІMV Ac-5017 − calcium (5 mM), N. vaccinii IMV B-7405 − calcium (5 and 10 mM), sodium (25−100 mM), potassium (50 and 100 mM). Additional introduction or increase the content of enzyme activators in cultivation medium of studied strains was accompanied by increasing activity of NADP+-dependent glutamate dehydrogenase in 1.5-3 times compared with that in the base medium.

The possibility of regulating biological properties of final product during cultivation of surfactants producer are discussed. The obtained results suppose modification of microbial surfactants characteristics with the change in cultivation medium of cations content − activators and/or inhibitors of key enzymes biosynthesis of component surfactants responsible for specific biological properties. 

1. Santos D. K., Rufino R. D., Luna J. M., Santos V. A., Sarubbo L. A. Biosurfactants: multifunctional biomolecules of the 21st century. International Journal of Molecular Sciences, 2016, vol. 17, no. 3. DOI: 10.3390/ijms17030401

2. Fair R. J., Tor Y. Antibiotics and bacterial resistance in the 21st century. Perspectives in Medicinal Chemistry, 2014, vol. 6, pp. 25−64. DOI: 10.4137/PMC.S14459

3. Demain A. L. Importance of microbial natural products and the need to revitalize their discovery. Journal of Industrial Microbiology & Biotechnology, 2014, vol. 41, no. 2, pp. 185–201. DOI: 10.1007/s10295-013-1325-z

4. Fracchia L., Banat J. J., Cavallo M., Ceresa C., Banat I. M. Potential therapeutic applications of microbial surfaceactive compounds. AIMS Bioengineering, 2015, vol. 2, no. 3, pp. 144–162. DOI: 10.3934/bioeng.2015.3.144

5. Robbel L., Marahiel M. A. Daptomycin, a bacterial lipopeptide synthesized by a nonribosomal machinery. Journal of Biological Chemistry, 2010, vol. 285, no. 36, pp. 27501–27508. DOI: 10.1074/jbc.R110.128181. Epub. 2010

6. Pirog T. P., Konon A. D, Skochko A. B. Microbial surface active substances use in biology and medicine. Biotekhnolohiya [Biotechnology], 2011, vol. 4, no. 2, pp. 24−38 (in Russian).

7. Pirog Т. P., Panasyuk E. V., Nikityuk L. V., Iutinska G. O. Influence of cultivation conditions on antimicrobial properties of Nocardia vaccinii ІMV B-7405 surfactants. Biotechnologia acta, 2016, vol. 9, no. 1, pp. 38−47. DOI: 10.15407/biotech9.01.038

8. Pirog T. P., Savenko I. V., Shevchuk T. A., Krutous N. V., Iutynska G. O. Antimicrobial properties surfactants synthesized under different cultivation conditions of Acinetobacter calcoaceticus ІMV B-7241. Microbiologichny Zhurnal [Microbiology Journal], 2016, vol. 78, no. 3, pp. 2−12 (in Russian).

9. Pirog T., Sofilkanych A., Konon A., Shevchuk T., Ivanov S. Intensification of surfactants’ synthesis by Rhodococcus erythropolis IMV Ac-5017, Acinetobacter calcoaceticus IMV B-7241 and Nocardia vaccinii K-8 on fried oil and glycerol containing medium. Food and Bioproducts Processing, 2013, vol. 91, no. 2, pp. 149−157. http://dx.doi.org/10.1016/j.fbp.2013.01.001.

10. Tareq F. S., Lee M. A., Lee H. S., Lee J. S., Lee Y. J., Shin H. J. Gageostatins A–C, antimicrobial linear lipopeptides from a marine Bacillus subtilis. Marine Drugs, 2014, vol. 12, no. 2, pp. 871–885. DOI: 10.3390/md12020871

11. Abalos A., Pinazo A., Infante M. R., Casals M., García F., Manresa A. Physicochemical and antimicrobial properties of new rhamnolipids produced by Pseudomonas aeruginosa AT10 from soybean oil refinery wastes. Langmuir, 2001, vol. 17, no. 5, pp. 1367−1371.

12. Mandal S. M., Barbosa A. E., Franco O. L. Lipopeptides in microbial infection control: scope and reality for industry. Biotechnology Advances, 2013, vol. 31, no. 5, pp. 338−345. DOI: 10.1016/j.biotechadv.2013.01.004

13. Podgorskiy V. S., Iutinskaya G. О., Pirog Т. P. Intensification of microbial synthesis technologies. Kyiv, Naykova Dumka [Scientific Thought], 2010. 327 p. (in Russian).

14. Pirog T. P., Shevchuk T. A., Mashchenko O. Yu., Parfenyuk S. A., Iutinskaya G. A. Effect of growth factors and some microelements on biosurfactant synthesis of Acinetobacter calcoaceticus IMV B-7241. Microbiologichny Zhurnal [Microbiology Journal], 2013, vol. 75, no. 5, pp. 19−27 (in Russian).

15. Pirog T. P., Shevchuk T. A., Beregova K. A., Kudrya N. V. Peculiarities of glucose and glycerol metabolism in Nocardia vaccinii IMB B-7405. Ukrainian Biochemical Journal, 2015, vol. 87, no. 2, pp. 66−75 (in Russian).

16. Sakamoto N., Kotre A. M., Savageau M. A. Glutamate dehydrogenase from Escherichia coli: purification and properties. Journal of Bacteriology, 1975, vol. 124 , no. 2, pp. 775−783.

17. Hudson R. C., Ruttersmith L. D., Daniel R. M. Glutamate dehydrogenase from the extremely thermophilic archaebacterial isolate AN1. Biochimica et Biophysica Acta, 1993, vol. 1202, no. 2, pp. 244–250.

18. Lee M. K., González J. M., Robb F. T. Extremely thermostable glutamate dehydrogenase (GDH) from the freshwater archaeon Thermococcus waiotapuensis: cloning and comparison with two marine hyperthermophilic GDHs. Extremophiles, 2002, vol. 6, no. 2, pp. 151–159.

19. Bhuiya M. W., Sakuraba H., Kujo C., Nunoura-Kominato N., Kawarabayasi Y., Kikuchi H., Ohshima T. Glutamate dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix K1: enzymatic characterization, identification of the encoding gene, and phylogenetic implications. Extremophiles, 2000, vol. 4, no. 6, pp. 333–341.

20. Sarada K. V., Rao N. A., Venkitasubramanian T. A. Isolation and characterisation of glutamate dehydrogenase from Mycobacterium smegmatis CDC 46. Biochimica et Biophysica Acta, 1980, vol. 615, no. 2, pp. 299–308.

21. Lin H. P. P., Reeves H. C. Purification and characterization of NADP+-specific glutamate dehydrogenase from Escherichia coli. Current Microbiology, 1991, vol. 22, no. 6, pp. 371–376.

22. Choudhury R., Punekar N. S. Aspergillus terreus NADP-glutamate dehydrogenase is kinetically distinct from the allosteric enzyme of other Aspergilli. Mycological Research, 2009, vol. 113, no. 10, pp. 1121–1126. DOI: 10.1016/j.mycres.2009.07.009