Abstract
Background
The invertebrate type (i-type) lysozyme not showing a clear homology with the known types of lysozyme was first demonstrated from a marine bivalve, conch and earthworm by N-terminal sequence. An i-type lysozyme isolated from the earthworm found to be up-regulated upon bacterial challenge, suggesting this lysozyme to function as an inducible immune factor. However, information on the i-type lysozyme related with digestive function is very limited in the earthworm.
Objective
The objective of this study is to investigate the molecular characteristics and function of the new i-type lysozyme from the earthworm.
Methods
To identify a new i-type lysozyme, multiple amino acid sequence alignment and phylogenetic analyses were employed. Its mRNA expression pattern was observed by fluorescent in situ hybridization (FISH).
Results
A new i-type lysozyme (Ea-iLys) from an earthworm, Eisenia andrei with the open reading frame of 678 bp (226 amino acid residues) appeared to comprise conserved 14 cysteine residues for disulfide bridges and amino acid residues for the enzyme activities of lysozyme and isopeptidase, of which mRNA expression is mainly localized in the lining of midgut epithelium. No significant expression signal was detected in immune competent sites such as chloragogue tissue, typhlosole region, body coelom and muscle layers.
Conclusion
Our results suggest that this enzyme primarily acts as a digestive enzyme rather than an innate immune factor.


Similar content being viewed by others
Explore related subjects
Discover the latest articles and news from researchers in related subjects, suggested using machine learning.References
Cho SJ, Valles Y, Weisblat DA (2014) Differential expression of conserved germ line markers and delayed segregation of male and female primordial germ cells in a hermaphrodite, the leech Helobdella. Mol Biol Evol 31:341–354
Cotuk A, Dales RP (1984) Lysozyme activity in the coelomic fluid and coelomocytes of the earthworm Eisenia foetida sav. in relation to bacterial infection. Comp Biochem Physiol A 78(3):469–474
Edwards CA, Fletcher KE (1988) Interactions between earthworms and microorganisms in organic-matter breakdown. Agric Ecosyst Environ 24:235–247
Fiołka M, Zagaja MP, Hułas-Stasiak M, Wielbo J (2012) Activity and immunodetection of lysozyme in earthworm Dendrobaena veneta (Annelida). J Invertebr Pathol 109(1):83–90
Ito Y, Yoshikawa A, Hotani T, Fukuda S, Sugimura K, Imoto T (1999) Amino acid sequences of lysozymes newly purified from invertebrates imply wide distribution of a novel class in the lysozyme family. Eur J Biochem 259:456–461
Jolles P (1996) Lysozymes: model enzymes in biochemistry and biology. Birkhauser Verlag, Basel
Jolles J, Jolles P (1975) The lysozyme from Asterias rubens. Eur J Biochem 54(1):19–23
Joskova R, Silerova M, Prochazkova P, Bilej M (2009) Identification and cloning of an invertebrate-type lysozyme from Eisenia andrei. Dev Comp Immunol 33:932–938
Lemos FJ, Terra WR (1991) Digestion of bacteria and the role of midgut lysozyme in some insect larvae. Comp Biochem Physiol B 100(2):265–268
Liu H, Wang J, Mao Y, Liu M, Niu S, Qiao Y, Su Y, Wang C, Zheng Z (2016) Identification and expression analysis of a new invertebrate lysozyme in Kuruma shrimp (Marsupenaeus japonicas). Fish Shellfish Immunol 49:336–343
Markad VL, Adav SS, Ghole VS, Sze SK (2016) Proteomics study revealed altered proteome of Dichogaster curgensis upon exposure to ash fly. Chemosphere 160:104–113
Park IY, Cha JR, Ok S-M, Shin C, Kim J-S, Kwak H-J, Yu Y-S, Kim Y-K, Medina B, Cho S-J, Park SC (2017) A new earthworm cellulase and its possible role in the innate immunity. Dev Comp Immunol 67:476–480
Ursic Bedoya RJ, Mitzey AM, Obraztsova M, Lowenberger C (2005) Molecular cloning and transcriptional activation of lysozyme-encoding cDNAs in the mosquito Aedes aegypti. Insect Mol Biol 14(1):89–94
Van Herreweghe JM, Michiels CM (2012) Invertebrate lysozymes: diversity and distribution, molecular mechanism and in vivo function. J Bisci 37(2): 327–348
Wang X, Xiaoqin L, Sun Z (2018) iTRAQ-based quantitative proteomic analysis of the earthworm Eisenia fetida response to Escherichia coli O157:H7. Ecotoxicol Environ Saf 160:60–66
Xue QG, Itoh N, Schey KL, Li YL, Cooper RK, La Peyre JF (2007) A new lysozyme from the eastern oyster (Crassostrea virginica) indicates adaptive evolution of i-type lysozymes. Cell Mol Life Sci 64(1):82–95
Xue QG, Hellberg ME, Schey KL, Itoh N, Eytan RI, Cooper RK (2010) A new lysozyme from the eastern oyster, Crassostrea virginica, and a possible evolutionary pathway for i-type lysozymes in bivalves from host defense to digestion. BMC Evol Biol 10:213–229
Acknowledgements
This research was supported by the Chung-Ang University Research Scholarship Grants in 2012–2013 and by “Cooperative Research Program for Agriculture Science and Technology Development [Project No.: PJ011661]” Rural Development Administration, Republic of Korea.
Author information
Authors and Affiliations
Corresponding authors
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
Yu, YS., Lee, JY., Woo, JW. et al. Identification and expression pattern of a new digestive invertebrate-type lysozyme from the earthworm. Genes Genom 41, 367–371 (2019). https://doi.org/10.1007/s13258-018-0776-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13258-018-0776-z


