Zvyagintseva T.N., Bakunina I.Yu., Alexeeva S.A., Ivanova E.P., Sova V.V., Burtseva Yu.V., Yermakova S.P., Nedashkovskaya O.I., Pivkin M.V., Mikhailov. - презентация

1 Zvyagintseva T.N., Bakunina I.Yu., Alexeeva S.A., Ivanova E.P., Sova V.V., Burtseva Yu.V., Yermakova S.P., Nedashkovskaya O.I., Pivkin M.V., Mikhailov V.V. Laboratory of Enzymic Chemistry Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok Laboratory of Enzymic Chemistry Pacific Institute of Bioorganic Chemistry, FEB RAS, Vladivostok

2 1,3;1,6- -D-Glucans a e dc b - 3,6-О-di-substituted Glc residue - nonsubstituted Glc residue - 3-О-substituted Glc residue - 6-О-substituted Glc residue a bc from Laminaria cichorioides a bc from Fucus evanescens Soloveva T.F., Elyakova L.A., Zvyagintseva T.N., Yermak I.M. Marine Technol. Soc : 35

3 Fucoidans - 3-О-substituted Fuc residue - 4 -О-substituted Fuc residue from Fucus evanescens (Bilan et al., Carbohydr.Res. 2002, 337: 719) Alginic Acids G-block М-block from Laminaria cichorioides (Zvyagintseva et al., J.Exp.Mar.Biol.Ecol., 2002 (in press) Zvyagintseva et al., Carbohydr. Res : 32.

4 Endo- and exo-1,3- -D-Glucanases Alginases Fucoidan hydrolases Endo-1,6- -D-Glucanases Cellulases - and -D-Glucosidases - and -D-Glucosidases - and -D-Galactosidases - and -D-Galactosidases - and -D-Mannosidases - and -D-Mannosidases -L-Fucosidases -L-Fucosidases N-Аc- - and N-Аc- -D-Glucosaminidases Sulfatases

5 Representatives of genera Acremonium, Aphanocladium, Mucor and Verticillium did not show any activity towards substrates used. There was no fucoidan hydrolase activity in all fungi tested. Representatives of genera Acremonium, Aphanocladium, Mucor and Verticillium did not show any activity towards substrates used. There was no fucoidan hydrolase activity in all fungi tested.

6 P.issachenkovii P.citrea P.nigrifaciens Glycosyl hydrolases Profiles of Some Pseudoalteromonas Ivanova, E.P. et al. Pseudoalteromonas issachenkonii sp. nov., a bacterium that degrades the thallus of the brown alga Fucus evanescens.//Int J Syst Evol Microbiol (Pt 1)

7 Biosynthesis of glycosyl hydrolases in bacteria degrading Fucus evanescens (experimental conditions) Fucus evanescens Fucus evanescens + inhibitor Ivanova E.P. et al. Two species of culturable bacteria associated with degradation of brown algae Fucus evanescens.//Microbial Ecology, 2002,v.43, N2, p

8 Cultivation P. issachenkovii, strain Effect of medium composition on protein synthesis and glycosyl hydrolases activities Cultivation P. issachenkovii, strain Effect of medium composition on protein synthesis and glycosyl hydrolases activities Alexeeva et al. Letters in Applied Microbiology 2002, 35: 1

9 - Fucoidan - Fucoidan + КММ 3296 (F.evanescens) - Fucoidan + КММ 3297 (Apostichopus japonica) - Fucoidan + КММ 3298 (Chorda fillum) Absorbamce at 490 nm Elution volume, ml Bakunina et al. Microbiology. 2002, 71: 41.

10 Re-chromatography Gel-filtration on BioGel Р-200 Ion-exchange chromatography Burtseva et al. Marine Biotechnology (in press)

11 Marine organisms are the valuable sources of the biologically active polysaccharides, the enzymes with unique specificity involved in their transformation and inhibitors of these enzymes. So, endo-1,3- -D-glucanases of marine organisms, found at first as hydrolases, catalyze three reactions running simultaneously and practically with almost equal efficiency. S''-A (reaction of transglycosylation) (reaction of transglycosylation) S' ESESESES E+SE+SE+SE+S ES'' S''-ОН (reaction of hydrolysis) S''-S (glucanosyl transferase activity) A S H2OH2O Sova V.V. et al. Biochemistry (Moscow) : 1300.

12 1,3;1,6- -D-Glucans a e dc b - 3,6-О-di-substituted Glc residue - nonsubstituted Glc residue - 3-О-substituted Glc residue - 6-О-substituted Glc residue a bc from Laminaria cichorioides a bc from Fucus evanescens Soloveva T.F., Elyakova L.A., Zvyagintseva T.N., Yermak I.M. Marine Technol. Soc : 35

13 Bacunina et al. Biochemistry (Moscow).1998, 63: 1420; 2002, 67: 830. Gal (1 3)Gal (1 4)GlcNAc (1 3)Gal (1 4)Glc-ceramide 2 1 Fuc B(III) Gal (1 4)GlcNAc (1 3)Gal (1 4)Glc-ceramide 2 1 Fuc O(H) -Galactosidase

14 I - Sea of Okhotsk; II - Sea of Japan; III - Indian Ocean; IV - South-Chinese Sea I - Sea of Okhotsk; II - Sea of Japan; III - Indian Ocean; IV - South-Chinese Sea Bacunina et al. -Galactosidase of the marine bacterium Pseudoalteromonas sp. KMM 701. Biochemistry (Moscow).1998, 63: 1420.

15 Phylogenetic tree