Abstract
Objective
In this study, we characterised a novel lysophospholipase (LysoPL) from the L. mucosae LM1 strain. The gene, LM-lysoPL, encoding LysoPL from L. mucosae LM1 was cloned, analyzed, and expressed.
Results
LM-lysoPL contained a conserved region and catalytic triad motif responsible for lysophospholipase activity. After purification, UHPLC-MS analysis showed that recombinant LM-LysoPL hydrolyzed phosphatidic acid, generating lysophosphatidic acid. The enzyme had greater hydrolytic activity against C16 and C18 fatty acids, indicating a preference for long-chain fatty acids. Enzymatic assays showed that the optimal pH and temperature of recombinant LM-LysoPL were 7 and 30 °C, respectively, and it was enzymatically active within a narrow pH range.
Conclusions
To the best of our knowledge, this is the first study to identify and characterize a lysophospholipase from lactic acid bacteria. Our findings provide a basis for understanding the probiotic role of L. mucosae LM1 in the gut.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Aoki J (2004) Mechanisms of lysophosphatidic acid production. Semin Cell Dev Biol 15:477–489. https://doi.org/10.1016/j.semcdb.2004.05.001
Balolong MP, Chae JP, Kang DK (2016) Expression and characterisation of neopullulanase from Lactobacillus mucosae. Biotechnol Lett 38:1753–1760. https://doi.org/10.1007/s10529-016-2152-x
Bathena SP, Huang J, Nunn ME et al (2011) Quantitative determination of lysophosphatidic acids (LPAs) in human saliva and gingival crevicular fluid (GCF) by LC–MS/MS. J Pharm Biomed Anal 56:402–407. https://doi.org/10.1016/j.jpba.2011.05.041
Brenner S (1988) The molecular evolution of genes and proteins: a tale of two serines. Nature 334:528–530. https://doi.org/10.1038/334528a0
Casas-Godoy L, Duquesne S, Bordes F et al (2012) Lipases: an overview. In: Methods in molecular biology, pp 3–30
Chaucheyras-Durand F, Durand H (2010) Probiotics in animal nutrition and health. Benef Microbes 1:3–9. https://doi.org/10.3920/BM2008.1002
Chen SCA, Wright LC, Santangelo RT et al (1997) Identification of extracellular phospholipase B, lysophospholipase, and acyltransferase produced by Cryptococcus neoformans. Infect Immunol 65:405–411
Cui Z, Wang Y, Pham BP et al (2012) High level expression and characterization of a thermostable lysophospholipase from Thermococcus kodakarensis KOD1. Extremophiles 16:619–625. https://doi.org/10.1007/s00792-012-0461-0
Deng W, Tsukahara ES et al (2007) The lysophosphatidic acid type 2 receptor is required for protection against radiation-induced intestinal injury. Gastroenterology 132:1834–1851. https://doi.org/10.1053/j.gastro.2007.03.038
Doi O, Nojima S (1975) Lysophospholipase of Escherichia coli. J Biol Chem 250:5208–5214
Haas E, Stanley DW (2007) Phospholipases. In: xPharm: the comprehensive pharmacology reference, pp 1–3
Hsu KH, Wang SY, Chu FH, Shaw JF (2010) Characterization and heterologous expression of a novel lysophospholipase gene from Antrodia cinnamomea. J Appl Microbiol 108:1712–1722. https://doi.org/10.1111/j.1365-2672.2009.04569.x
Hung ND, Kim MR, Sok D-E (2010) Purification and characterization of lysophospholipase C from pig brain. Neurochem Res 35:50–59. https://doi.org/10.1007/s11064-009-0029-2
Kiss GN, Fells JI, Gupte R et al (2012) Virtual screening for LPA2-specific agonists identifies a nonlipid compound with antiapoptotic actions. Mol Pharmacol. https://doi.org/10.1124/mol.112.079699
Kovačić F, Granzin J, Wilhelm S et al (2013) Structural and functional characterisation of TesA—a novel lysophospholipase A from Pseudomonas aeruginosa. PLoS ONE 8:e69125. https://doi.org/10.1371/journal.pone.0069125
Lee S-J, Leoni G, Neumann P-A et al (2013) Distinct phospholipase C-isozymes mediate lysophosphatidic acid receptor 1 effects on intestinal epithelial homeostasis and wound closure. Mol Cell Biol 33:2016–2028. https://doi.org/10.1128/MCB.00038-13
Ljungh A, Wadström T (2006) Lactic acid bacteria as probiotics. Curr Issues Intest Microbiol 7:73–90
Million M, Angelakis E, Paul M et al (2012) Comparative meta-analysis of the effect of Lactobacillus species on weight gain in humans and animals. Microb Pathog 53:100–108. https://doi.org/10.1016/j.micpath.2012.05.007
Richmond GS, Smith TK (2011) Phospholipases A1. Int J Mol Sci 12:588–612. https://doi.org/10.3390/ijms12010588
Salvetti E, Torriani S, Felis GE (2012) The genus Lactobacillus: a taxonomic update. Probiotics Antimicrob Proteins 4:217–226. https://doi.org/10.1007/s12602-012-9117-8
Schrag JD, Li Y, Wu S, Cygler M (1991) Ser-His-Glu triad forms the catalytic site of the lipase from Geotrichum candidum. Nature 351:761–764. https://doi.org/10.1038/351761a0
Sugimoto H, Hayashi H, Yamashita S (1996) Purification, cDNA cloning, and regulation of lysophospholipase from rat liver. J Biol Chem 271:7705–7711. https://doi.org/10.1074/jbc.271.13.7705
Sugimoto S, Sugimoto H, Aoyama C et al (2006) Purification and characterization of lysophospholipase D from rat brain. Biochim Biophys Acta 1761:1410–1418. https://doi.org/10.1016/j.bbalip.2006.09.013
Valdes AM, Walter J, Segal E, Spector TD (2018) Role of the gut microbiota in nutrition and health. BMJ 361:k2179. https://doi.org/10.1136/bmj.k2179
Valeriano VD, Parungao-Balolong MM, Kang DK (2014) In vitro evaluation of the mucin-adhesion ability and probiotic potential of Lactobacillus mucosae LM1. J Appl Microbiol 117:485–497. https://doi.org/10.1111/jam.12539
Valeriano VD, Bagon BB, Balolong MP, Kang DK (2016) Carbohydrate-binding specificities of potential probiotic Lactobacillus strains in porcine jejunal (IPEC-J2) cells and porcine mucin. J Microbiol 54:510–519. https://doi.org/10.1007/s12275-016-6168-7
Valeriano VDV, Oh JK, Bagon BB et al (2019) Comparative genomic analysis of Lactobacillus mucosae LM1 identifies potential niche-specific genes and pathways for gastrointestinal adaptation. Genomics 111:24–33. https://doi.org/10.1016/j.ygeno.2017.12.009
Yun CC, Kumar A (2015) Diverse roles of LPA signaling in the intestinal epithelium. Exp Cell Res 333:201–207. https://doi.org/10.1016/j.yexcr.2014.11.013
Zhu H, Liu P, Du J et al (2017) Identification of lysophospholipase protein from Spiroplasma eriocheiris and verification of its function. Microbiology 163:175–184. https://doi.org/10.1099/mic.0.000407
Acknowledgements
The present research was conducted by the research fund of Dankook University in 2019.
Author information
Authors and Affiliations
Corresponding author
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Kim, S.H., Song, J.H., Kim, J. et al. Characterisation of a lysophospholipase from Lactobacillus mucosae. Biotechnol Lett 42, 1735–1741 (2020). https://doi.org/10.1007/s10529-020-02895-0
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10529-020-02895-0