Russulaceae FAMILY MUSHROOMS LECTINS: FUNCTION, PURIFICATION, STRUCTURAL FEATURES and POSSIBILITIES of PRACTICAL APPLICATIONS L

Home , Lactarius deliciosus, Lactarius deterrimus, Lactarius lignyotus, Russula, Russulaceae, Russula delica

Reviews

UDC 615.32:547.96:582.284 https://doi.org/10.15407/biotech12.01.029 Russulaceae FAMILY MUSHROOMS LECTINS: FUNCTION, PURIFICATION, STRUCTURAL FEATURES AND POSSIBILITIES OF PRACTICAL APPLICATIONS L. V. PANCHAK

Danylo Halytsky Lviv National Medical University, Ukraine

E-mail: [email protected] Received 29.10.2018 Revised 17.11.2018 Accepted 14.01.2019 The purpose of this paper was to analise the results of own author’s research and literature date that concerned lectins of Russulaceae family mushrooms, which, despite their widespread, are still poorly investigated. Most studies merely reported about the determination of hemagglutinating activity and the isolation of pure lectin preparations from fresh fruit bodies of this family mushrooms. This article provides information about lectins physiological role in mushrooms, the list of Russulaceae family spesies tested for hemagglutinating activity, as well as procedure of purification, molecular structure and carbohydrate specificity of isolated lectins. Particulary, the most effective methods of lectins purification of the Russulaceae family were highlighted. High lability of the lectins molecules explaned the loss of the activity of these lectins during purification from raw material when standard procedures were applied, as well as the reason why these lectins were not obtained from the dried fruit bodies. Finally, practical application of lectins of Russulaceae family mushrooms in medical- biological researches are described.

Key words: le ctins, Russulaceae, hemagglutinating activity, carbohydrate specificity.

Lectins are a class of multivalent able quickly accumulate significant biomass, carbohydrate-binding proteins of non-immune which can be used for obtaining pure lectins. origin, which recognize various carbohydrate- Our attention was attracted to the lectins of containing structures with a high degree of Russulaceae family mushrooms, which, despite stereospecificity without catalytic activity [1, 2]. widespread occurrence of these mushrooms, are The carbohydrate specificity of sugars and poorly investigated at present [10]. On the basis glycoproteins is the cause of selective binding of own research and studies of other authors, of lectins to certain types of cells and tissues of this review attempts to explain the reason for living organisms [3]. this fact and outline the potential for practical Lectins specificity towards cells (associated application of Russulaceae family lectins. with the structure of glycans on their surface) manifests in the ability to agglutinate certain Function cells, such as erythrocytes belonging to a The family Russulaceae with two genera — particular human or animal phenotypes [4], to Lactarius (the milk-caps) and Russula (the precipitate glycans and cause various biological brittlegils), has a significant species diversity effects [5, 6]. (there are 750 species of genus Russula and Lectins were found in all kingdoms of 450 species of genus Lactarius documented living organisms: viruses, bacteria, plants, worldwide [11]), and consequently, a large mushrooms and animals [2, 7]. The vast majority number of potential sources of biologically of commercially available lectins are obtained active substances. from plants, but lectins of true fungi also are of Almost all specimens of this family form great interest [8, 9]. Their fruiting bodies are mycorrhizal associations with trees and

29 BIOTECHNOLOGIA ACTA, V. 12, No 1, 2019 shrubs, particularly with the main forest tree which were stored in herbarium for 8–9 years! species (pine, beech, oak, spruce, fir, etc.) [12, The intensity of agglutination reached 1: 13]. Lectins are involved in the mycorrhizal 64 for the L. cremor, L. quietus, L. piperatus symbiosis [14]. extracts, and 1: 32 for the L. torminosus, The physiological role of lectins in fungi L. controversus, L. thejogalus [21]. is associated with the specific recognition of Lactarius and Russula species of glycosylated structures at the level of cells, the Ukrainian Carpathians also have tissues and the whole organism. Lectins in hemagglutinating activity toward human mushrooms participate in the mobilization erythrocytes. Though the extracts of and transportation of sugars, in the process investigeted fungi did not show group of forming fruiting bodies (the formation of specificity, rabbit, dog and guinea pig primordia), in the creation of mycorrhiza and erythrocytes showed approximately the same the infection process (penetration of parasitic activity. Horse and dog erythrocytes were fungi into the host organism) [15]. often more sensitive to Russulaceae lectins, The fungal lectins are involved in the while cow and goat erythrocytes frequently recognition of the symbionts during the showed low sensitivity to these lectins [22, 23]. early stages of mycorrhizae formation. In So, no blood group-specific lectin among the Lactarius deterrimus-spruce model, the Lactarius and Russula species was found. Only facts that there is a lectin at the surface of the R. queletii exhibited specificity to pigeon and mycelial hyphae and the presence of specific horse erythrocytes [18]. sites for the lectin on the roots hairs are We have noticed a significant variability evidence of the involvement of the lectin in in titers of hemagglutination, depending the recognition of the host spruce. Thus, the on the age of fruiting bodies, weather and morphologically very similar L. deterrimus, temperature conditions, even within one L. deliciosus and L. salmonicolor are associated mycelium (measurements were performed on with the the spruce (Picea), pine (Pinus), and mycelia of L. torminosus and L. pergamenus). the fir (Abies), respectively with a remarkable Activity may differ more than in 250 times! specificity [16]. Although clear correlation between the age of mushroom and the titer of hemagglutination Detection was not found, in general, younger Detection of lectins in mushroom mushrooms had higher hemagglutinating extracts is most often carried out using activity than older ones [22, 24]. Evidently, hemagglutination inhibition assay with human it is much easier to detect lectins in extracts and animal erythrocytes, method can be found at the moment of their highest activity. in reference [2, 17]. According to our observations, higher A large number of basidiomycetes were activity of lectins correlates with their higher screened using the hemagglutination test. content. Particularly, among the 104 species, collected Other researchers, in particular [18, 19] in Poland, aqueous extracts from L. rufus came to the same conclusion that caps of young and L. vellereus showed high agglutination carpophores are usually richer in lectins. activity and were among ten the most active. The extract from R. cyanoxanta had distinctly Purification higher agglutination activity at 4 C, than that Lectins are proteins or glycoproteins that obtained at room temperature [18]. consist of one or more polypeptide chains. Agglutinins were well represented Procedures that are customary for purification by Lactarius species among 403 British other proteins (precipitation with salts, ion- higher fungi, but fewer by Russula species exchange, affinity and gel chromatography, (Table 1) [19]. However, in a study of 110 preparative electrophoresis, etc.) can be used species of Japanese fungi only R. emetica, to purified them [1, 2]. However, unlike one of six Russula species, did not exhibit lectins of other basidiomycota mushrooms, hemagglutination. All species that were active they have certain features. This is due to the against human erythrocytes also agglutinated high sensitivity of Russolaceae family lectins rabbit red blood cells, except R. violeipes to pH changes, salt precipitation and organic that agglutinated only human cells [19–20]. solvents, drying and freezing of raw materials. It’s interesting that the ability to At the same time, these lectins, can usually agglutinate human erythrocytes of all ABO withstand heating to 6570 С [22, 23]. This groups had dried mushroom caps of 21 species should be taken into consideration when of the genus Lactarius (out of 48 studied), purifying them.

30 Reviews

Table 1. List of Russulaceae family spesies tested for hemagglutinating activity

No Lactarius DC. ex S. F. Gray: References No Russula (Fr.) S. F. Gray: References 1 L. aurantiacus Fr. [21] 1 R. adusta (Pers.) Fr. ** [19] 2 L. blennius (Fr.) Fr. [21,19] 2 R. aeruginea Lindblad [19, 23] R. alpina (A. Blytt) F. H. 3 L. controversus (Fr.) Fr. [21,19] 3 [19] Møller & Jul. Schäff ** R. atropurpurea (Krombh.) 4 L. cremor Fr. [21] 4 [19, 23] Britzelm. R. carminea (J. Schaeff.) 5 L. decipiens Quel. [21] 5 [19] Kühner & Romagn. 6 L. deliciosus (L.) Gray [18, 21, 25, 19, 22 ] 6 R. claroflava Grove [19] R. cyanoxantha (Jul. Sch ff.) 7 L. deterrimus Gr ger [26,19] 7 ä [19, 23] ö Fr. 8 L. flavidulus S. Imai [27] 8 R. delica Fr. [32] R. emetica (Schaeff.: Fr.) S.F. 9 L. fluens Boud. [19] 9 [18] Gray ⃰ 10 L. glaucescens Cossl. [19] 10 R. farinipes Romell. [19] R. flavida Frost &Peck apud 11 L. glyciosmus Fr. [21] 11 [20] Peck 12 L. helvus Fr. [21] 12 R. foetens (Fr.) Fr. [23] 13 L. laeticolor (Imai) Imaz. [20] 13 R. grisea (Pers. ex Secr.) Fr. [19, 23] 14 L. lignyotus Fr. [28,21] 14 R. illota Romagn. [19] L. necator (Bull. ex Fr.) 15 [22] 15 R. laurocerasi Melzer [19, 20] Karst. 16 L. pergamenus (Fr.) Fr. [21, 22, 24] 16 R. lepida [33] R. nigricans (Bull. ex Merat) 17 L. piperatus (Scop.) Gray [21, 19, 29, 22 ] 17 [19, 23] Fr. 18 L. porninsis Roll. [21] 18 R. paludosa Britzelm. [19] 19 L. pubescens Fr. [21] 19 R. queletii Fr. [18, 19] 20 L. quietus (Fr.) [21, 19, 20, 22] 20 R. rosacea (Pers.) S. F. Gray [20] R. sardonia Fr.em Romagn. 21 L. rufus (Scop.) Fr. [18, 19, 30, 22] 21 [19] *** 22 L. ruginosus Romagn. [19] 22 R. subfoetens Smith [19] L. salmonicolor R. Heim & 23 [21, 31] 23 R. vesca Fr. [19] Leclair 24 L. sanguifluus Paul ex Fr. [21] 24 R. violeipes Quel. [20] 25 L. serifluus DC.ex Fr. [21] 26 L. subvellereus Peck [20] 27 L. subzonarius Hongo [20] 28 L. tabidus Fr. [19] 29 L. thejogalus (Bull.) Fr. [21] L. torminosus (Schaeff.) 30 [21, 19, 30, 22] Gray 31 L. turpis (Weinm.)** [19] 32 L. vellereus (Fr.) Fr. [18, 21, 19, 22] 33 L. vietus (Fr.) Fr. [19] 34 L. volemus (Fr.) Fr. [19, 22] L. zonarioides K hner 35 [21] & Romagn

* Hemagglutination only with trypsin treated erythrocytes ** Hemagglutination only with bromelin treated erythrocytes *** Hemagglutination with neuraminidase treated erythrocytes

31 BIOTECHNOLOGIA ACTA, V. 12, No 1, 2019

The titres of haemagglutination in sap from L. torminosus and L. pergamenus within one mycelium

Although many Russulaceae family species Affinity chromatography is the most showed agglutination activity there are effective method for the purification of the only several publications on obtained lectins lectins. The affinity sorbents used for their (Table 2). isolation were: stromas of erythrocytes embedded Not always the hemagglutinating activity in polyacrylamide gel [25, 26, 31], copolymer of Russulaceae lectins is remained entirely by of polyvinyl alcohol with a blood group specific the precipitation of lectins with ammonium substance [30, 34, 29, 24], fetuin immobilized on sulfate and ethanol, which are most often used Sepharose [28]. The purification of other lectins in the purification of protein molecules [22, (L. flavidulus, R. delica, R. foetens, R. lepida) [27, 23]. 32, 23, 33] was carried out by a combination of It should be noted that all authors purified ion-exchange chromatography and gel filtration. lectins only from raw materials harvested However, in our opinion, these methods are fresh. None of the cited studies didn’t explain more labour-intensive and less productive than why these lectins were not obtained from dried affinity chromatography. fruit bodies. In own research [22, 23], it has Lectins were eluted from affinity sorbents been shown that the hemagglutination activity either with 0.1 M borate buffer (pH 9.0) strongly reduces or completely disappears [28] or heated to +65 С 1 M NaCl. Specific after drying, as well as freezing of most species carbohydrate was not used to remove the lectin of this family mushrooms. from a column due to the absence of interaction The reason for this, in our opinion, is of these lectins with monosaccharides and low- the loss of activity of these lectins during cost di- and oligosaccharides. purification from raw material using standard The elution of lectins by acidic buffer is procedures. not recommended because it can cause loss of

32 Reviews

Table 2. List of isolated lectins of Russulaceae family species Structural Refe- Species Isolation Sugar Specificity Properties rences L. lignyotus By affinity chromatogra- 98 kDa, Desialyzed fetuin, desialyzed [28] phy on Sepharose 4B con- Tetrameric with glycoprotein from edible bird’s taining immobilized 22 kDa subunits nest and desialyzed mucin from fetuin porcine submaxillary glands

L. deliciosus* By a combination of affin- 37 kDa, Dimeric All the lectins are most specific [25] ity chromatography on with two types for DGal1-3DGlcNAc residues stromas of group O eryth- of subunits (TF antigen) rocytes embedded in poly- (about 19,000 acrylamide gel and and 18,000) hydroxylapatite and gel filtration chromatography L. deterrimus* 37 kDa, Dimeric [26] * Isolated from the carpophores, lectins are also expressed at the surface of the hyphae

L. salmonicolor* 39 kDa, Dimeric [31]

L. rufus By affinity chromatogra- 98 kDa, Group-specific substances from [30] phy on copolymer of poly- Hecameric with human blood erythrocytes, vinyl alcohol with a blood 17 ± 1 kDa sub- asialo-BSM, asialo-ovomucoid, group B specific substance units human and bovine thyroglobu- lins, orosomucoid, fetuin, transferrin, human Ig G, -phenyl N-acetyl-D- glucosaminopyranoside, 4-nitrophenyl--D- galactopyranoside L. torminosus 98 kDa, Fetuin, asialo-BSM, BSM, group- [34] Hecameric with specific substances from human 17 ± 1 kDa sub- blood erythrocytes, human Ig G, units transferrin

L. piperatus 97±3 kDa Group-specific substances from [29] human blood erythrocytes, human Ig G

L. pergamenus By a combination of etha- 96 kDa, The lectin weakly interacts with [24] nol precipitation, affinity Hecameric DGalNAc, while DGal1- chromatography on copo- with 16 kDa 3DGalNAc and DGal1-3DGlcNAc lymer of polyvinyl alcohol subunits are the most probable candidates and human blood B group for ligands, with which the lectin specific substance, and ion interacts at disaccharides level. exchange chromatography Among them fetuin of fetal calf on DEAE-Toyopearl serum and group-specific substances A, B, and H of human blood were the strongest

33 BIOTECHNOLOGIA ACTA, V. 12, No 1, 2019

Table 2. Сontinued Structural Refe- Species Isolation Sugar Specificity Properties rences L. flavidulus The chromatographic proce- 29.8 kDa, Lactose, p-nitrophenyl -D- [27] dure utilized comprised Dimeric glucopyranoside, p-nitrophe- anion-exchange chromatog- nyl -D-glucopyranoside, ino- raphy on DEAE-cellulose, sitol and inulin cation-exchange chromatography on CM-cellulose, anion-exchange chromatography on SP-Sepharose and gel filtration by fast protein liquid chromatography on Superdex 75 R. delica It was adsorbed on both 60 kDa, Dimeric Inulin and o-nitrophenyl--D- [32] SP-Sepharose and with 30 kDa galactopyranoside Q-Sepharose and unadsorbed subunits on DEAE-cellulose

R. foetens By ion exchange chromato- Not done Asialo-ovomucoid [23] graphy on CM-cellulose

R. lepida The purification scheme 32 kDa, Inulin and o-nitrophenyl--D- [33] involved (NH4)2SO4 precipi- Dimeric with 16 galactopyranoside tation, ion exchange chro- kDa subunits matography on DEAE- cellulose and SP-Sepharose, and fast protein liquid chromatography-gel filtration on Superdex 75 their activity. In particular, as our studies Molecular structure have shown [22, 23], Russulaceae lectins are There is a small amount of data in the quite enough stable at high temperatures, but literature concerning the deep learning are very sensitive to changes in pH: even a on molecular structure of lectins of the short-term reduction of pH to 4.5 often leads Russulaceae family mushrooms. In particular, to a loss of 50–75% of hemagglutinating there is no data on the crystallographic activity, and at pH 3.5 activity of many (but structure of lectins from mushrooms of this not all) species was completely lost. That’s family in complex with a specific carbohydrate why the elution of almost all Lactarius lectins inhibitor. from the affinity column was carried out According to the literature (Table 2), using 1 M NaCl solution heated to +65 С. An purified lectins consist of two (L. deterrimus, attempt to eluate the L. deterrimus lectin by L. salmonicolor, L. flavidulus, R. delica 0.1 M solution of N-acetyl-D-galactosamine, and R. lepida) or four identical subunits although was successful [26], did not allow (L. lignyotus), L. deliciosus lectin is also the authors to obtain the expected activity dimer, but has non-identical subunits, with a of lectin. In their opinion it was due to molecular weight of 18 and 19 kDa. incomplete unbounding of this carbohydrate Except the L. lignyotus lectin, whose from the lectin by dialysis. subunits are bound by disulfide bridges, other The yield of lectins from fresh mushroom studied lectins of the Russulaceae family have biomass varied from 3 mg/kg in L. perga- subunits bound noncovalently. menus [24], 6.4 mg/kg in L. deterrimus [26] to According to our research, L. rufus and  30 mg/kg in L. deliciosus [25]. L. torminosus lectins consist of six subunits with a molecular weight of about 17 kDa,

34 Reviews and L. pergamenus — about 16 kDa. It was be said of the L. torminosus, L. rufus and determined that the linkage between the L. pergamenus lectins, the best sugars to individual subunits is very labile and the interact with were complex oligosaccharides lectin molecule can easily disintegrate, with minimal active disaccharide chains of possibly gradually, which is accompanied by DGal1-3DGlcNAc or DGal1-3DGalNAc. loss of haemagglutinating activity. Herewith The best inhibitors of their activity were the hexamer possesses the highest specific group-specific blood components human haemagglutinating activity. Disintegration of (ovariomucin) and human immunoglobulin G the lectin molecule is taking place under the [34, 30, 24]. action of minor pH changes and even under In case of L. lignyotus lectin the most the action of such soft factors as freezing effective inhibitors of its agglutinating activity or precipitation of the lectin-containing were desialized fetuin, desialized glycoprotein fraction of proteins with ammonium sulfate, from edible bird’s nest and desialyzed mucin not to mention about drying of mushrooms at from porcine submaxillary glands [28]. +52 ± 2 C [22, 23]. It should be noted that Inhibitors of agglutinating activity of hexameric structure of a molecule is the R. delica and R. lepida lectins were inulin and phenomenon quite rare in lectins; not more o-nitrophenyl--D galactopyranoside [32, 33]. than ten such molecules are described, the most famous is a lectin from an edible snail Practical application (Helix pomatia) [1, 2]. The selectivity of lectins to carbohydrate The data obtained by us explain the decrease structures makes them very important tools of haemagglutinating activity by changes in for biochemical studying of glycoconjugates of the molecular structure of fungal lectins of the the membrane and cell wall [35]. Thereby, due Russulaceae family, observed during drying to the high specificity of binding of lectins to and storage of the fruit bodies and throughout membrane carbohydrate structures, they have the purification of lectins. The disappearance become extremely useful for the identification of the hemagglutinating activity is due to and differentiation of closely related species the high lability of the quaternary structure or strains of single-celled parasitic organisms. of the lectin molecule; this fact probably also In particular, L. controversus and R. nigricans explains a small number of studies on lectins of lectins characterized the variety of sugar a given family. However, lectins of the genus structures of the cell walls of 114 pathogenic Lactarius, as compared with lectins of the strains of the Candida genus. The lectin from genus Russula, are generally more resistant to L. deliciosus was used in researching the biology such impacts, particularly towards drying, and and taxonomy of the fungal organisms from therefore more promising for further research. the class of Chytridiomycetes inhabiting the alimentary canal in ruminants. L. torminosus Carbohydrate specificity lectin was used in researching glycoconjugates The most important functional taking part in the identification reactions of characteristic of lectins is their specificity to embryonic cells in the urogenital morphogenesis carbohydrates, which often determines the in bird embryos [15]. opportunities for their further use. Lectins from R. delica and R. lepida potently The literature suggests that Russulaceae inhibited proliferation of HepG2 hepatoma and family lectins rarely exhibit specificity to MCF 7 breast cancer cells [32, 33]. The lectin monosaccharides, and much more often to from L. flavidulus suppressed the proliferation complex oligosaccharide structures, at least of HepG2 hepatoma and L1210 leukemic to disaccharides. Mostly such disaccharide is cells [27]. Both above mentioned lectins DGal1-3DGalNAc (TF antigen) [25, 22, 23]. from L. flavidulus and R. delica inhibited the The hemagglutinating activity of activity of human immunodeficiency virus L. flavidulus lectin was inhibited by a variety type 1 (HIV-1) reverse transcriptase [27, 32]. of simple sugars, in particular lactose, The lectin of L. deliciosus, which is specific p-nitrophenyl -D- and -D-glucopyranosides, to DGal1-3DGalNAc, can be used to follow inositol, and by the polysaccharide inulin the expression the tumor-associated Thomsen- [27]. The L. deterrimus, L. deliciosus and Friedenreich (TF) antigen [36]. L. salmonicolor lectins have almost identical The purified by us lectins from molecular mass and the same affinity for L. torminosus and L. pergamenus may find DGal1-3DGalNAc, a disaccharide which application as the histochemical reagents for contains D-galactose and N-acetyl-D- the comparative histochemical investigation of galactosamine [25, 26, 31]. The same can kidneys of the newborn and adult rats [37, 38].

35 BIOTECHNOLOGIA ACTA, V. 12, No 1, 2019

The key role of lectins in the formation our opinion, is the features of the molecular of mycorrhiza between certain conifer tree structure. The molecules of these lectins species and Russulaceae family mushroom consist of 2, 4 or 6 subunits, and the species has been proved. linkage between the individual subunits is The most effective method for the very labile and the molecule of lectins can purification of lectins from the Russulaceae easily disintegrate (possibly gradually), is affinity chromatography using sorbents which goes hand in hand with the loss of with DGal1-3DGlcNAc structures haemagglutinating activity. (immobilized fetuin, blood group-specific Lectins isolated from widespread substances). In the purification process Russulaceae family mushrooms can find should be taken into account high sensitivity practical applications for the research of of Russulaceae lectins to changes in pH and mycorrhizal symbiosis, as histochemical precipitation with ammonium sulfate and reagents in the detection of carbohydrate ethanol, their relatively thermal stability (up structures (for example, TF antigen), for to + 70 C). the study of the topography of membranes The reason for the relative instability and glycoconjugates of cell walls of various of the lectins of the Russulaceae family, in biological objects.

REFERENCES 1. Sharon N. Lectins: carbohydrate-specific 12. Ryabchuk V. P., Kolodiy T. V. The systematic reagents and biological recognition molecules. state of the order Russulales species. J. Biol. Chem. 2007, 282 (5), 2753–2764. Naukovyi visnyk NLTU Ukrainy. 2010, 2. Antonyuk V. O. Lectins and their resources. 20 (12), 8–13. (In Ukrainian). Lviv: Quart. 2005, 554 p. (In Ukrainian). 13. Smith S. E., Read D. J. Mycorrhizal 3. Gabius H. J., Roth J. An introduction to the Symbiosis, Third edition. London: Academic sugar code. Histochem. Cell Biol. 2017, 147 (2), Press. 2008, 815 p. 111–117. 14. Nikitina V. E., Loshchinina E. A., 4. Khan F., Khan R. H., Sherwani A., Mohmood S., Vetchinki na E. P. Lectins from Mycelia of Azfer M. A. Lectins as markers for blood Basidiomycetes. Int. J. Mol. Sci. 2017, 18 (7), grouping. Med. Sci. Monit. 2002, 8 (12), 13–34. 293–300. 15. Konska G. Lectins of higher fungi 5. Kumar K. K., Chandra K. L., Sumanthi J., (macromycetes)-their occurrence, Reddy G. S., Shekar P. C., Reddy B. Biological physiological role, and biological activity. role of lectins: A review. J. Orofacial Sci. Int. J. Med. Mushrooms. 2006, 8 (1), 2012, 4 (1), 20–25. 19–30. 6. Hamid R., Masood A., Wani I. H., Rafiq S. 16. Giollant M., Guillot J., Damez M., Dusser M., Lectins: Proteins with Diverse Applications. Didier P., Didier E. Characterization of а J. Appl. Pharmaceut. Sci. 2013, 3 (4), S93–S103. lectin from Lactarius deterrimus (Research 7. Santos A. F. S., da Silva M. D. C., Napoleão T. H., on the possible involvement of the fungal Paiva P. M. G., Correia M. T. S., Coelho L. C. B. B. lectin in recognition between mushroom and Lectins: Function, structure, biological spruce during the early stages of mycorrhizae properties and potential applications. Curr. formation). Plant Physiol. 1993, 101 (2), Top. Peptide Prot. Res. 2014, V. 15, P. 41–62. 513–522. 8. Singh S. S., Wang H., Chan Y. S., Pan W., Dan X., 17. Sano K., Ogawa H. Hemagglutination Yin C. M., Akkouh O., Ng T. B. Lectins from edible (inhibition) assay. Meth. Mol. Biol. 2014, mushrooms. Molecules. 2015, 20 (1), 446–469. V. 1200, P. 47–52. 9. Hassan M. A., Rouf R., Tiralongo E., May T. W., 18. Konska G. A survey of domestic species of Tiralongo J. Mushroom lectins: specificity, Basidiomycetes fungi for the presence of structure and bioactivity relevant to human lectins in their carpophores. Acta Societatis disease. Int. J. Mol. Sci. 2015, 16 (4), Botanicorum Poloniae. 1988, 57 (2), 7802–7838. 247–260. 10. Singh R. S., Bhari R., Kaur H. P. Mushroom 19. Pemberton R. T. Agglutinins (lectins) from lectins: Current status and future some British higher fungi. Mycol. Res. 1994, perspectives. Crit. Rev. Biotechnol. 2010, 98 (3), 277–290. 30 (2), 99–126. 20. Yagi F., Sakai T., Shiraishi N., Yotsumoto M., 11. Atri N. S., Sharma S., Saini M. K., Das K. Mukoyoshi R. Hemagglutinins (lectins) Researches on Russulaceous Mushrooms- in fruit bodies of Japanese higher fungi. AnAppraisal. Kavaka. 2016, V. 47, P. 63–82. Mycoscience. 2000, 41 (4), 323–330.

36 Reviews

21. Rymkiewicz A. Phythämagglutinine (Lekline) 30. Panchak L. V., Antonyuk V. O. Purification in Pilzen aus der Gattung Lactarius. Acta of lectin from fruiting bodies of Lactarius Mycol. 1988 (1989), 24 (1), 59–64. rufus (Scop.: Fr.) Fr. and its carbohydrate 22. Panchak L. V., Benzel L. V. Investigation of specificity. Ukr. Biokhim. Zh. 2007, V. 2, P. lectins of Russulaceae family mushrooms. 123–127. (In Ukrainian). Genus Lactarius. Pharm. Zh. 2010, V. 3, 31. Guillot J., Giollant M., Damez M., Dusser M. P. 99–105. (In Ukrainian). Intervention des lectines fongiques dans les 23. Panchak L. V., Benzel L. V. Investigation of événements précoces de reconnaissance arbre/ lectins of Russulaceae family mushrooms. champignon au cours de la formation des Genus Russula. Pharm. Zh. 2009, V. 3, ectomycorhizes. Acta Botanica Gallica. 1994, P. 115–121. (In Ukrainian). 141 (4), 443–447. 24. Panchak L. V., Antonyuk V. O. Purification 32. Zhao S., Zhao Y., Li S, Zhao J., Zhang G., of a lectin from fruit bodies of Lactarius Wang H., Ng T. B. A novel lectin with highly pergamenus (Fr.) Fr and studies its potent antiproliferative and HIV-1 reverse properties. Biochemistry (Moscow). 2011, transcriptase inhibitory activities from the 76 (4), 438–449. edible wild mushroom Russula delica. Prot. 25. Guillot J., Giollant M., Damez M., Dusser M. Pept. Lett. 2010, 27 (2), 259–265. Isolation and characterization of a lectin 33. Zhang G. G., Sun J., Wang H., Ng T. B. First from the mushroom Lactarius deliciosus. isolation and characterization of a novel J. Biochem. (Tokyo). 1991, 109 (6), 840–845. lectin with potent antitumor activity from 26. Giollant M., Guillot J., Damez M., Dusser M., a Russula mushroom. Phytomedicine. 2010, Didier P., Didier E. Characterization of а 17 (10), 775–781. lectin from Lactarius deterrimus (Research 34. Panchak L. V. Materials of scientific-practical on the possible involvement of the fungal conference with international participation lectin in recognition between mushroom and “Medicines to people. Modern problems spruce during the early stages of mycorrhizae of creation, study and approbation of formation). Plant Physiol. 1993, 101 (2), medicines”. Kharkiv, Ukraine. 2007, P. 297– 513–522. 303. (In Ukrainian). 27. Wu Y., Wang H., Ng T. B. Purification 35. Sybirna N. O., Buslyk T. V. Glycoproteins of and characterization of a lectin with erythrocyte membranes and the structure antiproliferative activity toward cancer of their carbohydrate determinants. Studia cells from the dried fruit bodies of Lactarius Biologica. 2009, 3 (1), 79–93. (In Ukrainian). flavidulus. Carbohydr. Res. 2011, 346 (16), 36. Guillot J., Konska G. Lectins in Higher Fungi. 2576–2581. Biochem. Systematics Ecol. 1997, 25 (3), 203–230. 28. Sychrova H., Ticha M., Kocourek J. Studies 37. Ambarova N. O., Panchak L. V., Antonyuk R. V. on lectins. LIX. Isolation and properties of Materials of VI scientific-practical conference lectins from fruiting bodies of Xerocomus with international participation of students and chrysenteron and Lactarius lignyotus. Can. young scientists “Scientific potential of youth – J. Biochem. Cell Biol. 1985, 63 (7), 700–704. progress of medicine of the future”. Uzhhorod, 29. Panchak L. V., Gulko R. М. Materials of Ukraine. 2008, P. 165–166. (In Ukrainian). All-Ukrainian scientific and practical 38. Ambarova N. O., Panchak L. V., Antonyuk R. V. conference of students and young scientists Collection of materials of scientific-practical “Actual questions of development of new conference “Applied aspects of morphology of medicines”. Kharkiv, Ukraine. 2008, P. 80. experimental and clinical research”. Ternopil, (In Ukrainian). Ukraine. 2008, P. 3. (In Ukrainian).

37 BIOTECHNOLOGIA ACTA, V. 12, No 1, 2019

ЛЕКТИНИ ГРИБІВ ЛЕКТИНЫ ГРИБОВ РОДИНИ Russulaceae: СЕМЕЙСТВА Russulaceae: ФУНКЦІЇ, ОЧИЩЕННЯ, ФУНКЦИИ, ОЧИСТКА, СТРУКТУРНІ ОСОБЛИВОСТІ СТРУКТУРНЫЕ ОСОБЕННОСТИ ТА МОЖЛИВОСТІ ПРАКТИЧНОГО И ВОЗМОЖНОСТИ ПРАКТИЧЕСКОГО ЗАСТОСУВАННЯ ПРИМЕНЕНИЯ

Л. В. Панчак Л. В. Панчак

Львівський національний медичний Львовский национальный медицинский університет ім. Данила Галицького университет им. Данилы Галицкого

Метою роботи є опис результатів влас- Целью работы является описание резуль- них досліджень та досліджень інших авто- татов собственных исследований и исследо- рів, що стосуються лектинів грибів родини ваний других авторов, касающихся лектинов Russulaceae, які, незважаючи на велике по- грибов семейства Russulaceae, которые, несмо- ширення, на сьогодні недостатньо досліджені. тря на широкое распространение, сегодня не- Більшість робіт повідомляє лише про визна- достаточно исследованы. Большинство работ чення гемаглютинувальної активності та виді- сообщает лишь об определении гемагглюти- лення лектинових препаратів зі свіжих плодо- нирующей активности и выделении лектино- вих тіл грибів даної родини. вых препаратов из сырых плодовых тел грибов У статті наведено інформацію про фізіо- данного семейства. логічну роль лектинів у грибах, перелік видів В статье приведена информация о физио- грибів родини Russulaceae, протестованих на логической роли лектинов в грибах, перечень гемаглютинувальну активність, а також проце- видов грибов семьи Russulaceae, протестиро- дуру очищення, структуру молекули та вугле- ванных на гемагглютинирующую активность, водну специфічність виділених лектинів. Роз- а также о процедуре очистки, структура моле- глянуто найбільш ефективні методи очищення кулы и углеводной специфичности выделен- лектинів родини Russulaceae. ных лектинов. Рассмотрены наиболее эффек- Висока лабільність молекули пояснює тивные методы очистки лектинов семейства втрату активності цих лектинів під час очи- Russulaceae. щення із сировини з використанням стан- Высокая лабильность молекулы объясняет дартних процедур, а також неможливість їх потерю активности этих лектинов при очистке отримання із сушених плодових тіл. Також из сырья с использованием стандартных проце- описано практичне застосування лектинів дур, а также невозможность их получения из грибів родини Russulaceae в медико-біологіч- сушеных плодовых тел. Также описано прак- них дослідженнях. тическое применения лектинов грибов семей- ства Russulaceae в медико-биологических ис- следованиях. Ключові слова: лектини, Russulaceae, гема- глютинувальна активність, вуглеводна Ключевые слова: лектины, Russulaceae, специфічність. гемагглютинирующая активность, углеводная специфичность .