Mikrobiol. Z. 2021; 83(3):35-45.
doi: https://doi.org/10.15407/microbiolj83.03.035

Effect of Different Ligand and Different Ligand Heterometal Xylaratohermanates
on the Activity of α-L-Rhamnosidases Eupenicillium erubescens,
Cryptococcus albidus and Penicillium tardum

O.V. Gudzenko1, N.V. Borzova1, L.D. Varbanets1, I.I. Seifullina2, O.A. Chebanenko2, O.E. Martsinko2

1Zabolotny Institute of Microbiology and Virology, NAS of Ukraine
154 Akad. Zabolotny Str., Kyiv, 03143, Ukraine

2Mechnikov Odessa National University
2 Dvoryanskaya Str., Odessa, 65029, Ukraine

α-L-Rhamnosidase [EC], enzyme of the hydrolase family has a wide range of applications: in the food industry, for example, in winemaking to improve the quality and aroma of wines, in the production of citrus juices and drinks to remove bitter components (naringin) that improves the quality and nutritional value of these products; in research as an analytical tool for studying the structure of complex carbohydrate-substituted biopolymers. For the successful use of α-L-rhamnosidases in various biotechnological processes, an important aspect is the development of ways to increase their activity. The main factors affecting the growth and metabolism of microorganisms, including the synthesis of enzymes, are the physicochemical conditions of cultivation, the composition of the nutrient medium, the introduction of substances that raise the yield of the enzyme, which is manifested in an increase in its activity. At present, one of the priority directions of modern research is the study of the effect of various effector compounds that are capable to modify the studied enzymatic activity. In this work, which is a continuation of previous studies, a number of mixed-ligand and mixed-ligand-different-metal coordination germanium compounds of with xylaric acid (H5Xylar), 1,10-phenanthroline (Phen), 2,2-bipyridine (bipy) and ions of 3d-metals (Fe2+, Ni2+, Cu2+, Zn2+) were selected as effectors. Study of the effect of these complexes on the activity of Eupenicillium erubescens, Cryptococcus аlbidus and Penicillium tardum α-L-rhamnosidases were the aim of this work. Methods. The objects of research were α-Lrhamnosidases from Eupenicillium erubescens 248, Cryptococcus albidus 1001, and Penicillium tardum IMV F-100074. The α-L-rhamnosidase activity was determined by the Davis method using naringin as a substrate. We used 12 coordination compounds of germanium as modifiers of enzyme activity, the composition and structure of which were established using a combination of physical and chemical research methods: elemental analysis, thermogravimetry, IR spectroscopy and X-ray structural analysis. Structures of seven compounds are deposited in the Cambridge Crystallographic Database. When studying the effect of various compounds on the activity of enzymes, concentrations of 0.1 and 0.01% were used, exposure times were 0.5 and 24 hours. The test compounds were dissolved in 0.1% dimethyl sulfoxide. UV-spectra of absorption of native and chemical modified preparations of the enzymes were studied by spectrophotometer-fluorimeter DeNovix DS-11 in the range of 220–340 nm, concentration of the enzyme preparation 1.0 mg of protein/mL. Results. Analysis of the totality of the obtained data (exposure time 24 h, concentration 0.1%) regarding the effect of the studied compounds on the activity of E. erubescens, C. albidus and P. tardum α-L-rhamnosidases showed that the influence of the studied modifiers for the activity of α-L-rhamnosidases varies depending on the producer strain. Our data allow us to present the following series of modifiers in accordance with an increase in their effect on the activity of enzymes of different producers: E. еrubescens: 12 < 11 < 5 < 3 < 4=10 < 1 < 3 < 8 < 2 < 6 < 7; C. albidus: 10 < 11 < 12 < 9 < 3 < 1=5 < 8=4 < 2 < 6 < 7; P. tardum: 12=2 < 3 < 4 < 11 < 5 < 8 < 1 < 9 < 6 < 10 < 7. Conclusions. The results obtained allow us to conclude that compound (7)(-tris(bipyridine) nickel(II) μ-dihydroxyxylaratogermanate(IV)) is the most effective activator of α-L-rhamnosidases of all three micromycete strains, compound (6)(tris(phenanthroline)nickel(II) μ-dihydroxyxylaratogermanate(IV)) − on α-L-rhamnosidase from E. erubescens and C. albidus, while compound (10)-(copper(II) μ-dihydroxyxylaratogermanate(IV)-cuprate(II)) − only of P. tardum α-L-rhamnosidase.

Keywords: Eupenicillium erubescens 248, Cryptococcus albidus 1001, Penicillium tardum ІМV F-100074, α-L-rhamnosidase, different ligand and different ligand heterometal xylaratohermanates, 1,10-phenanthroline, 2,2-bipyridine.

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  1. Yadav V, Yadav PK, Yadav S, Yadav KDS. Alpha-L-rhamnosidase: a review. Proc Biochem. 2010; 45(8):1226–1235. https://doi.org/10.1016/j.procbio.2010.05.025
  2. Gudzenko OV, Varbanets LD, Seifullina II, Martsinko EE, Pirozhok OV, Chebanenko EA. Germanium coordination compounds for increasing of α-L-rhamnosidase activity. Biotechnologia Acta. 2019; 12(4):19–26. https://doi.org/10.15407/biotech12.04.019
  3. Gudzenko OV, Varbanets LD, Seifullina II, Chebanenko EA, Martsinko EE, Afanasenko EV. The influence of coordinative tartrate and malatogermanate compounds on the activity of α-L-rhamnosidase preparations from Penicillium tardum, Eupenicillium erubescens and Cryptococcus albidus. Ukr Biochem J. 2020; 92(4):85–95. https://doi.org/10.15407/ubj92.04.085
  4. Varbanets LD, Matselyukh OV, Nidyalkova NA, Avdiyuk EV, Gudzenko EV, Seifullina II, Masanovets GN, Khitrich NV. [The coordination compounds of cobalt (II, III) with dithiocarbamic acid derivatives – modificators of hydrolytic enzymes activity]. Biotechnologia Acta. 2013; 6(1):73–80. Ukrainian. https://doi.org/10.15407/biotech6.01.073
  5. Seifullina II, Khitrich GN. Coordinative compounds of zinc with n-substituted thiocarbamoil-N′-pentamethylensulfenamides – activity modifiers of enzymes of proteolytic and glycolytic action. Ukr Biochem Z. 2011; 83(3):25–36.
  6. Varbanets LD, Gudzenko OV, Borzova NV. Rhamnosidase from Eupenicillium erubescens: purification and characterization. Nauka i Studia. 2013; 41(109):11–23.
  7. Borzova N, Gudzenko O, Varbanets L. Purification and characterization of a naringinase from Cryptococcus albidus. Applied biochemistry and biotechnology. 2018; 184(3):953–969. https://doi.org/10.1007/s12010-017-2593-2
  8. Gudzenko OV, Varbanets LD. [Purification and physico-chemical properties of α-L-rhamnosidase Penicillium tardum]. Mikrobiol Z. 2016; 78(1):13–22. Ukrainian. https://doi.org/10.15407/microbiolj78.01.013
  9. Davis DW. Determination of flavonones in citrus juice. Anal Biochem. 1947; 19:476–478. https://doi.org/10.1021/ac60007a016
  10. Chebanenko EA, Seifullina II, Martsinko EE, Dyakonenko VV, Shishkina SV. Directed Structure Formation in Tetranuclear Xylaratogermanates(IV) with Complex Phenanthrolinecopper(II) Cations. Russian Journal of Inorganic Chemistry. 2020; 65(11):1703–1711. https://doi.org/10.1134/S0036023620110029
  11. Chebanenko EA, Seifullina II, Martsinko EE, Dyakonenko VV, Shishkina SV. Synthesis and Structure of Frame Xylaratogermanate Salts with Protonated Phenanthroline and Its Complexes with Fe(III) and Ni(II) as Cations. Russian Journal of Inorganic Chemistry. 2019; 64(9):1132–1137. https://doi.org/10.1134/S0036023619090043
  12. Chebanenko EA, Martsinko EE, Seifullina II, Dyakonenko VV, Shishkina SV. Structural Features And Properties of Heteronuclear Germanium (IV) and Some 3d Metal Complexes with Xylaric Acid and 2,2′-Bipyridine. Journal of Structural Chemistry. 2018; 59(6):1462–1469. https://doi.org/10.1134/S0022476618060318
  13. Lucas LH, Ersoy BA, Kueltzo LA, Joshi SB, Brandau DT, Thyagarajapuram N, Peek LJ, Middaugh CR. Probing protein structure and dynamics by second-derivative ultraviolet absorption analysis of cation–π interactions. Protein Science. 2006; 15:2228–2243. https://doi.org/10.1110/ps.062133706
  14. Antosiewicz JM, Shugar D. UV-Vis spectroscopy of tyrosine side -groups in studies of protein structure. Part 2: selected applications. Biophys Rev. 2016; 8:163–177. https://doi.org/10.1007/s12551-016-0197-7
  15. Pramodini Devi S, Hemakumar Singh RK, Kadam RM. Synthesis and spectroscopic studies on copper(II) binuclear complexes of 1-phenylamidino-O-alkylurea (alkyl=n-propyl, n- and iso-butyl) with 1,3-diaminopropane or ethylenediamine. Inorg Chem. 2006; 45(5):2193–2918. https://doi.org/10.1021/ic051037t
  16. Debnath A, Hussain F, Masram DT. Synthesis, characterization and antifungal studies of metalloquinolone [Cd2(nal)2(phen)2(Cl)2]; Complex Metals. 2014; 1(1):96–102. https://doi.org/10.1080/2164232X.2014.889581
  17. Tezuka T, Higashino A, Akiba M, Nakamura T. Organogermanium (Ge-132) suppresses activities of stress enzymes responsible for active oxygen species in monkey liver preparation. A. E. R. 2017; 5(2):1323. https://doi.org/10.4236/aer.2017.52002
  18. Ali MM, Noaman E, Kamal S, Soliman S, Ismail DA. Role of germanium L-cysteine α-tocopherol complex as stimulator of some antioxidant defense systems in gamma-irradiated rats. Acta Pharm. 2007; 57:1–12. https://doi.org/10.2478/v10007-007-0001-0
  19. Singh SK, Das A. The n → π* interaction: a rapidly emerging non-covalent interaction. Phys Chem Chem Phys: PCCP. 2015; 17(15):9596–9612. https://doi.org/10.1039/C4CP05536E