Obtaining and characteristic of degradation products of Lactobacillus acidophilus K 3111 peptidoglycan


Keywords: peptidoglycan, muropeptides, disintegration, ultrasonic, enzymatic hydrolysis

Abstract

It is shown in the article that low molecular weight degradation products of peptidoglycans of bacterial walls possess high immunotropic activity and can stimulate evolutionarily fixed mechanisms of immune response. It is proposed to obtain low molecular weight fragments of peptidoglycans of cell walls of Lactobacillus acidophilus K 3111 – muropeptides. Fragmentation was carried out by combining physical and enzymatic methods of disintegration. As a physical factor of influence, an ultrasonic treatment was used, as enzymatic factor, a treatment with proteases of digestive, plant and microbial origin was used. These enzymes have a significant range of substrate specificity and can catalyze the specific binding of peptidoglycans to bacterial walls. The regularities of ultrasonic disintegration of Lactobacillus acidophilus K 311 has been investigated in the article. The rational conditions of ultrasonic disintegration are the treatment of biomass at a frequency of 35 kHz for 600 seconds. Under these conditions, the maximum amount of amino acids is accumulated in the disintegrate and the integrity of most bacterial cells is violated. The regularities of the enzymolysis of peptidoglycans of cell walls of Lactobacillus acidophilus K 3111 by pancreatin, papain and protosubtilin has been determined. It has been shown that ultrasonic disintegration, which precedes enzymatic hydrolysis, contributes to a significant increase in the content of low molecular weight peptides in hydrolysates by almost 50%. The expediency of using an enzyme of plant origin papain for enzymatic fragmentation of peptidoglycan is substantiated. When the ratio of the enzyme (papain): substrate 1: 200 and the duration of the process for 180 minutes, the highest content of low molecular weight peptides in hydrolysates (6.6 mg/cm3) is achieved. It is proved that the target compounds muropeptides are contained in the products of enzymatic hydrolysis, the content of muropeptides reaches 38% of the total amount of low molecular weight peptides at the hydrolysis by papain, 31% –by pancreatin and 23% – by protosubtilin.

References

Kapustyan, A.I., & Cherno, N.K. (2015). Perspektivyi ispolzovaniya biologicheski aktivnyih bakterialnyih gidrolizatov dlya nutritivnoy podderzhki naseleniya s rastroystvami immunnoy sistemyi. Pischevaya nauka i tehnologiya. 2(31), 18–25. doi: 10.15673/2073-8684.31/2015.44263 (in Russian).

Cherno, N., & Kapustyan, A. (2016). Immunological properties of the bacterial origin compounds. Food science and technology, 10(3), 19–28. doi: 10.15673/fst.v10i3.175.

Vavricka, S.R. (2004). hPepTl transports muramyl dipeptide, activating NF -kappaB and stimulating IL-8 secretion in human colonic Caco2/bbe cells. Gastroenterology. 127, 1401–1409.

Traub, S., Aulock, Von, S., Hartung, T., & Hermann, C. (2006). MDP and other muropeptides – direct and synergistic effects on the immune system. J Endotoxin Res. 12, 69–85. doi: 10.1179/096805106X89044.

Qingshan, Lv. et al. (2012). MDP Up-Regulates the Gene Expression of Type I Interferons in Human Aortic En-dothelial Cells. Molecules. 17, 3599–3608. doi: 10.3390/molecules17043599.

Matsui, K., Matsui, K., & Ikeda, R. (2014). Peptidoglycan in combination with muramyldipeptide synergistically induces an interleukin-10-dependent T helper 2-dominant immune response. Microbiol Immunol. 58, 260–265. doi: 10.1111/1348-0421.12139.

Kawai, T., & Akira, S. (2010). The role of pattern-recognition receptors in innate immunity: Update on Toll-like receptors. Nat. Immunol. 11, 373–384. doi: 10.1038/ni.1863.

Chapot-Chartier, M.P., & Kulakauskas, S. (2014). Cell wall structure and function in lactic acid bacteria. Microb Cell Fact. 13, 1–9. doi: 10.1186/1475-2859-13-S1-S9.

Matsumoto, S. et al. (2009). A component of polysaccharide peptidoglycan complex on Lactobacillus induced an improvement of murine model of inflammatory bow-el disease and colitis-associated cancer. Immunology. doi: 10.1111/j.1365-2567.2008.02942.x.

Ku¨hner, D., Stahl, M., Demircioglu, D.D., & Bertsche, U. (2014). From cells to muropeptide structures in 24 h: Peptidoglycan mapping by UPLC-MS. Sci. Rep. 4, 74–94. doi: 10.1038/srep07494.

Regulski, K. (2012). Analysis of the Peptidoglycan Hydrolase Complement of Lactobacillus casei and Characterization of the Major c-D-Glutamyl-L-Lysyl-Endopeptidase. PLoS ONE. 7(2), e32301. doi: 10.1371/journal.pone.0032301.

Gavrilin, M.V., Senchukova, G.V, & Senchenko, S.P. (2007). Vyibor optimalnyih usloviy polucheniya gidrolizatov molochnokislyih bakteriy termokislotnyim sposobom. Him.-farm. Zhurn. 41(2), 54–56 (in Russian).

Senchenko, S.P., Samoylov, V.A., Gostischeva, N.M, Senchukova, G.V., & Gavrilin, M.V. (2005). Izuchenie sostava preparata, poluchennogo na osnove gidrolizata molochnokislyih bakteriy. Him.-farmats. Zhurn. 39(3), 51–53 (in Russian).

Livinskaya, E.P., Kovalenko, N.K., & Garmasheva, I.L. (2011). Dezintegraciya laktobacill i ehnterokokkov dlya polucheniya fragmentov kletochnyh stenok. Mіkrobіologіchnij zhurnal. 73(3), 26–32 (in Russian).

Ovsyannikova, L.V., & Komarova, E.L. (2012). Sravnytel'naya kharakterystyka proteolytycheskykh fermentov rastytel'noho proyskhozhdenyya – papayna y bromelayna, Dietary supplements market. 7(74), 3 (in Russian).

Geiger, C., Spieb, T., Korn, S.M., Kötter, P., & Entian, K.-D. (2017), Specificity of subtilin-mediated activation of histidine kinase SpaK, Appl Environ Microbiol. 83. doi: 10.1128/AEM.00781-17.

Golovach, T.N., Gavrilenko, N.V., Zhabanos, N.K., & Kurchenko, V.P. (2008). Zakonomirnosti hidrolizu syrovatkovykh bilkiv ekzo- I endoproteaz. Works BGU of Biochemistry. 3(1), 1–15 (in Russian).

Yakubke, H.-D., & Eshkide, H. (1985), Aminokislotyi, peptidyi, belki. Trans. from gem. (in Russian).

Semak, I.V., Zyiryanova, T.N., & Gubich, O.I. (2007). Biohimiya belkov: praktikum dlya studentov biol. Fak. spets. 1-31 01 01 «Biologiya», Minsk: BGU (in Russian).

Morris, D.L. (1948). Quantitative Determination of Carbohydrates With Dreywood's Anthrone Reagent. Science, 107(2775), 254–255.

Shaphaev, E.G. Tsyirenov, V.Zh., & Chebunina, E.I. (2015). Dezintegratsiya kletok v biotehnologii. Uchebnoe posobie, VSGTU,Ulan-Ude in Russian).

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Published
2018-03-02
How to Cite
Kapustian, A., & Cherno, N. (2018). Obtaining and characteristic of degradation products of Lactobacillus acidophilus K 3111 peptidoglycan. Scientific Messenger of LNU of Veterinary Medicine and Biotechnologies. Series: Food Technologies, 20(85), 141-147. https://doi.org/10.15421/nvlvet8526