Article Versions
Export Article
Research Article
Open Access Peer-reviewed

Mundticin KS: Characterization and Production Method

Alena Abaimova1, Nikolay Kartsev1, Edvard Svetoch1, Olga Tazina1, Tatiana Novikova1, Mikhael Platonov2, Irina Mitsevich1, Maria Kanashenko1, Rostislav Zhumakaev3, Konstantin Detushev4, Marat Teymurazov1,

1Department of Molecular Microbiology, State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation, 142279

2Department of Highly Dangerous Infections, State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation

3Biotechnology Deparment, State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation

4Collection Cultures Deparment, State Research Center for Applied Microbiology and Biotechnology, Obolensk, Moscow Region, Russian Federation

Journal of Applied & Environmental Microbiology. 2021, 9(1), 9-21. DOI: 10.12691/jaem-9-1-3
Received June 02, 2021; Revised July 05, 2021; Accepted July 18, 2021

Abstract

The paper aims to develop a method for the single-stage purification of mundticin KS and to study its physicochemical and antimicrobial properties. Mundticin produced by strain Enterococcus mundtii B-8398 and belonging to class IIa bacteriocins was obtained by adding 10% of the CM Sephadex C-25 sorbent to the culture medium before cultivation. At the end of cultivation, the sorbent was collected and packed into a column followed by a one-step elution. As a result, a fraction containing mundticin KS was obtained with a purification quality of about 70%. The activity of mundticin KS remained stable over a wide range of pH and temperatures. It is completely inactivated by such enzymes as proteinase K and α-chymotrypsin and partially by trypsin; however, such enzymes as amylase, lipase, and papain do not have any effect on its activity. Mass spectral analysis, electrophoresis, and DNA sequencing showed the high range similarity of mundticin KS produced by strains E. mundtii B-8398 to other previously studied mundticins. The antibacterial activity of mundticin KS was recorded ageist all tested strains of Listeria spp., Enterococcus spp., and Clostridium perfringens in nanomolar concentrations. In addition, mundticin KS inhibits the growth of most tested Gram-positive food-borne pathogens. By the sorbent cultivation method, it was possible to increase the yield of mundticin KS by 2,9 times compared to the control.

Keywords:

bacteriocin, Mundticin KS, purification methods, antibacterial spectrum
[1]  Feng G, Guron GKP, Churey JJ, and Worobo RW (2009) Characterization of Mundticin L, a Class IIa Anti-Listeria Bacteriocin from Enterococcus mundtii CUGF08. Appl Environ Microbiol 75: 5708-5713.View Article  PubMed 
 
[2]  Ferreira AE, Canal N, Morales D, Fuentefria DB and Corção G (2007) Characterization of enterocins produced by Enterococcus mundtii isolated from human feces. Braz Arch Biol Techn 50: 249-258.View Article 
 
[3]  Minahk C.J., Farias M.E., Sesma F., Morero R.D (2000) Effect of Enterocin CRL35 on Listeria monocytogenes cell membrane. FEMS Microbiol Lett 192: 79-83.View Article  PubMed 
 
[4]  Nawaz F, Khan MN, Javed A, Ahmed I, Ali N, Ali MI, Bakhtiar SM, Imran M (2019) Genomic and Functional Characterization of Enterococcus mundtii QAUEM2808, Isolated From Artisanal Fermented Milk Product Dahi. Front Microbiol 10: 434.View Article  PubMed 
 
[5]  Zendo T, Eungruttanagorn N, Fujioka S, Tashiro Y, Nomura K, Sera Y, Kobayashi G, Nakayama J, Ishizaki A and Sonomoto K (2005) Identification and production of a bacteriocin from Enterococcus mundtii QU 2 isolated from soybean. J Appl Microbiol 99: 1181-1190.View Article  PubMed 
 
[6]  Bennik MH, Vanloo B, Brasseur R, Gorris LGM, Smid EJ. (1998). A novel bacteriocin with a YGNGV motif from vegetable-associated Enterococcus mundtii: full characterization and interaction with target organisms. BBA-Biomemranes 1373, Issue 1: 47-58.View Article 
 
[7]  Botes M, Loos B, van Reenen CA, Dicks LMT. (2008). Adhesion of the probiotic strains Enterococcus mundtii ST4SA and Lactobacillus plantarum 423 to Caco-2 cells under conditions simulating the intestinal tract, and in the presence of antibiotics and anti-inflammatory medicaments. Arch Microbiol 190: 573-584.View Article  PubMed 
 
[8]  Cui Y, Zhang C, Wang Y, Shi J, Zhang L, Ding Z, Qu X, Cui H (2012) Class IIa bacteriocins: diversity and new developments. Int J Mol Sci 13: 16668-16707.View Article  PubMed 
 
[9]  Drider D, Fimland G, Hechard Y, McMullen LM, Prevost H. (2006). The continuing story of class IIa bacteriocins, Microbiol Mol Biol Rev 70: 564-582.View Article  PubMed 
 
[10]  Comerlato CB, Buboltz JR, Santestevan NA, da Motta AS and Frazzon APG (2016) Antimicrobial compounds produced by Enterococcus spp. isolates from fecal samples of wild South American fur seals. J Microbiol Antimicrob 8(3): 14-21.View Article 
 
[11]  Kawamoto S, Shima J, Sato R, Eguchi T, Ohmomo S, Shibato J, Horikoshi N, Takeshita K, Sameshima T (2002) Biochemical and genetic characterization of Mundticin KS, an antilisterial peptide produced by Enterococcus mundtii NFRI 7393. Appl Environ Microbiol 68: 3830-3840.View Article  PubMed 
 
[12]  Patrzykat A, Friedrich CL., Zhang L, Mendoza V and Hancock REW (2002) Sublethal Concentrations of Pleurocidin-Derived Antimicrobial Peptides Inhibit Macromolecular Synthesis in Escherichia coli. Antimicrob Agents CH 46 No. 3: 605-614.View Article  PubMed 
 
[13]  Salvucci E., Saavedra L. and Sesma F (2007) Short peptides derived from the NH2-terminus of subclass IIa bacteriocin enterocin CRL35 show antimicrobial activity. J Antimicrob Chemoth 59: 1102-1108.View Article  PubMed 
 
[14]  Henning C, Gautam D, Muriana PM. (2015). Identification of multiple bacteriocins in Enterococcus spp. Using an Enterococcus-specific bacteriocin PCR Array. Microorganisms 3(1): 1-16.View Article  PubMed 
 
[15]  Hirose S, Igarashi K, Naba T, Tsuga Y and Minato A (1977) Studies on Salivary Gland Ribonucleases: II. Purification of Ribonucleases from Bovine Submaxillary Gland and the Effects of Polyamines on Their Activities. J Biochem 82 Issue 6: 1605-1612.View Article  PubMed 
 
[16]  Jackson CR, Fedorka-Cray PJ, Barrett JB. (2004). Use of a genus- and species-specific multiplex PCR for identification of Enterococci. Clin Microbiol 42(8): 3558-3565.View Article  PubMed 
 
[17]  Ennahar S, Sashihara T, Sonomoto K and Ishizaki A. (2000). Class IIa bacteriocin biosynthesis, structure and activity. FEMS Microbiol. Rev., 24: 85-106.View Article  PubMed 
 
[18]  Schagger H and von Jagow G (1987) Tricine-sodium dodecyl sulfatepolyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166: 368-379.View Article 
 
[19]  Alispahic M, Hummel K, Jandreski-Cvetkovic D, Nöbauer K, Razzazi-Fazeli E, Hess M, Hess C. (2010). Species-specific identification and differentiation of Arcobacter, Helicobacter and Campylobacter by full-spectral matrix-associated laser desorption/ionization time of flight mass spectrometry analysis. J Med Microbiol 59(Pt 3): 295-301.View Article  PubMed 
 
[20]  Fox A (2006) Mass spectrometry for species or strain identification after culture or without culture: Past, present, and future. J Clin Microbiol 44(8): 2677-2680.View Article  PubMed 
 
[21]  Sahl H. G., Brandis H (1981) Production, purification and chemical properties of an antistaphylococcal agent produced by Staphylococcus epidermidis. J Gen Microbiol 127: 377-384.View Article  PubMed 
 
[22]  Yang R, Johnson MC and Ray B (1992) Novel Method To Extract Large Amounts of Bacteriocins from Lactic Acid Bacteria. Appl Environ. Microbiol 58(10): 3355.View Article  PubMed