Asian-Australasian Journal of Animal Sciences

Asian Australasian Association of Animal Production Societies (아세아태평양축산학회)
권호 표지
DOI QR코드

DOI QR Code

Molecular Cloning and Characterization of a New C-type Lysozyme Gene from Yak Mammary Tissue

  • Jiang, Ming Feng (College of Life Science and Technology, Southwest University for Nationalities) ;
  • Hu, Ming Jun (College of Life Science and Technology, Southwest University for Nationalities) ;
  • Ren, Hong Hui (College of Life Science and Technology, Southwest University for Nationalities) ;
  • Wang, Li (Key Laboratory of Animal Genetics & Breeding, Southwest University for Nationalities, State Ethnic Affairs Commission and Ministry of Education)
  • Received : 2015.02.10
  • Accepted : 2015.05.11
  • Published : 2015.12.01
Download PDF
⟨ Previous Next ⟩

Abstract

Milk lysozyme is the ubiquitous enzyme in milk of mammals. In this study, the cDNA sequence of a new chicken-type (c-type) milk lysozyme gene (YML), was cloned from yak mammary gland tissue. A 444 bp open reading frames, which encodes 148 amino acids (16.54 kDa) with a signal peptide of 18 amino acids, was sequenced. Further analysis indicated that the nucleic acid and amino acid sequences identities between yak and cow milk lysozyme were 89.04% and 80.41%, respectively. Recombinant yak milk lysozyme (rYML) was produced by Escherichia coli BL21 and Pichia pastoris X33. The highest lysozyme activity was detected for heterologous protein rYML5 (M = 1,864.24 U/mg, SD = 25.75) which was expressed in P. pastoris with expression vector $pPICZ{\alpha}A$ and it clearly inhibited growth of Staphylococcus aureus. Result of the YML gene expression using quantitative polymerase chain reaction showed that the YML gene was up-regulated to maximum at 30 day postpartum, that is, comparatively high YML can be found in initial milk production. The phylogenetic tree indicated that the amino acid sequence was similar to cow kidney lysozyme, which implied that the YML may have diverged from a different ancestor gene such as cow mammary glands. In our study, we suggest that YML be a new c-type lysozyme expressed in yak mammary glands that plays a role as host immunity.

Keywords

References

  1. Bionaz, M. and J. J. Loor. 2008. Acsl1, agpat6, fabp3, lpin1, and slc27a6 are the most abundant isoforms in bovine mammary tissue and their expression is affected by stage of lactation. J. Nutr. 138:1019-1024. https://doi.org/10.1093/jn/138.6.1019
  2. Brundige, D. R., E. A. Maga, K. C. Klasing, and J. D. Murray. 2010. Consumption of pasteurized human lysozyme transgenic goats' milk alters serum metabolite profile in young pigs. Transgenic. Res. 19:563-574. https://doi.org/10.1007/s11248-009-9334-4
  3. Callewaert, L. and C. W. Michiels. 2010. Lysozymes in the animal kingdom. J. Biosci. 35:127-160. https://doi.org/10.1007/s12038-010-0015-5
  4. Cereghino, J. L. and J. M. Cregg. 2000. Heterologous protein expression in the methylotrophic yeast pichia pastoris. FEMS Microbiol. Rev. 24:45-66. https://doi.org/10.1111/j.1574-6976.2000.tb00532.x
  5. Corin, K., H. Pick, P. Baaske, B. L. Cook, S. Duhr, C. J. Wienken, D. Braun, H. Vogel, and S. Zhang. 2012. Insertion of t4- lysozyme (t4l) can be a useful tool for studying olfactoryrelated gpcrs. Mol. Biosyst. 8:1750-1759. https://doi.org/10.1039/c2mb05495g
  6. Gallagher, D. S. Jr., D. W. Threadgill, A. M. Ryan, J. E. Womack, and D. M. Irwin. 1993. Physical mapping of the lysozyme gene family in cattle. Mamm. Genome 4:368-373. https://doi.org/10.1007/BF00360587
  7. Hannig, C., B. Spitzmüller, W. Hoth-Hannig, and M. Hannig. 2011. Targeted immobilisation of lysozyme in the enamel pellicle from different solutions. Clin. Oral Investig. 15:65-73. https://doi.org/10.1007/s00784-009-0357-2
  8. He, A. X. and L. Li. 2004. Study on the relation between yak performance and ecological protection. In: Proceedings of the Fourth International Congress on Yak. September 20-26. Chengdu, China.
  9. Hermann, J., J. Jolles, D. H. Buss, and P. Jolles. 1973. Amino acid sequence of lysozyme from baboon milk. J. Mol. Biol. 79:587-595. https://doi.org/10.1016/0022-2836(73)90408-7
  10. Irwin, D. M. 2004. Evolution of cow nonstomach lysozyme genes. Genome 47:1082-1090. https://doi.org/10.1139/g04-075
  11. Jauregui-Adell, J. 1975. Heat stability and reactivation of mare milk lysozyme. J. Dairy Sci. 58:835-838. https://doi.org/10.3168/jds.S0022-0302(75)84646-7
  12. Jiang, M., Y. Chen, Y. Wang, J. J. Loor, Y. Ye, Y. Wen, X. Zi, Y. Cai, and J. K. Drackley. 2010. Yak (bos grunniens) stomach lysozyme: Molecular cloning, expression and its antibacterial activities. Anim. Biotechnol. 21:25-35.
  13. Jolles, J., E. M. Prager, E. S. Alnemri, P. Jolles, I. M. Ibrahimi, and A. C. Wilson. 1990. Amino acid sequences of stomach and nonstomach lysozymes of ruminants. J. Mol. Evol. 30:370-382. https://doi.org/10.1007/BF02101891
  14. Kastorna, A., V. Trusova, G. Gorbenko, and P. Kinnunen. 2012. Membrane effects of lysozyme amyloid fibrils. Chem. Phys. Lipids 165:331-337. https://doi.org/10.1016/j.chemphyslip.2012.02.002
  15. Lan, F., H. Hu, W. Jiang, K. Liu, X. Zeng, Y. Wu, and Z. Gu. 2012. Synthesis of superparamagnetic fe3o4/pmma/sio2 nanorattles with periodic mesoporous shell for lysozyme adsorption. Nanoscale 4:2264-2267. https://doi.org/10.1039/c2nr12125e
  16. Lin, J. L., R. C. Ruaan, and H. J. Hsieh. 2007. Refolding of partially and fully denatured lysozymes. Biotechnol. Lett. 29:723-729. https://doi.org/10.1007/s10529-007-9320-y
  17. Marsh, M. B. and C. D. Rice. 2010. Development, characterization, and technical applications of a fish lysozyme-specific monoclonal antibody (mab m24-2). Comp. Immunol. Microbiol. Infect. Dis. 33:e15-23. https://doi.org/10.1016/j.cimid.2009.10.002
  18. Ng, A., M. Heynen, D. Luensmann, and L. Jones. 2012. Impact of tear film components on lysozyme deposition to contact lenses. Optom. Vis. Sci. 89:392-400. https://doi.org/10.1097/OPX.0b013e31824c0c4a
  19. Prieur, D. J. 1986. Tissue specific deficiency of lysozyme in ruminants. Comp. Biochem. Physiol. B 85:349-353. https://doi.org/10.1016/0305-0491(86)90011-8
  20. Proctor, V. A. and F. E. Cunningham. 1988. The chemistry of lysozyme and its use as a food preservative and a pharmaceutical. Crit. Rev. Food Sci. Nutr. 26:359-395. https://doi.org/10.1080/10408398809527473
  21. Steinhoff, U. M., B. Senft, and H. M. Seyfert. 1994. Lysozymeencoding bovine cdnas from neutrophile granulocytes and mammary gland are derived from a different gene than stomach lysozymes. Gene 143:271-276. https://doi.org/10.1016/0378-1119(94)90109-0
  22. Tamura, K., D. Peterson, N. Peterson, G. Stecher, M. Nei, and S. Kumar. 2011. Mega5: Molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol. Biol. Evol. 28:2731-2739. https://doi.org/10.1093/molbev/msr121
  23. Wang, C. S. and H. U. Kloer. 1984. Purification of human lysozyme from milk and pancreatic juice. Anal. Biochem. 139:224-227. https://doi.org/10.1016/0003-2697(84)90409-3
  24. Wang, N., Y. Wang, G. Li, N. Sun, and D. Liu. 2011. Expression, characterization, and antimicrobial ability of t4 lysozyme from methylotrophic yeast hansenula polymorpha a16. Sci. China Life Sci. 54:520-526. https://doi.org/10.1007/s11427-011-4174-x
  25. White, F. H. Jr., H. A. McKenzie, D. C. Shaw, and R. J. Pearce. 1988. Studies on a partially purified bovine milk lysozyme. Biochem. Int. 16:521-528.
  26. Wilken, L. R. and Z. L. Nikolov. 2011. Process evaluations and economic analyses of recombinant human lysozyme and hen egg-white lysozyme purifications. Biotechnol. Prog. 27:733-743. https://doi.org/10.1002/btpr.593
  27. Zhang, P., Y. Wang, M. Jiang, L. Zhu, J. Li, M. Luo, H. Ren, and L. Liu. 2014. Molecular cloning, recombinant protein expression, tissue distribution and functional analysis of a new c-type lysozyme from lezhi black goat rumen. Can. J. Anim. Sci. 94:27-34. https://doi.org/10.4141/cjas2012-150

Cited by

  1. Purification and characterization of lysozyme from Chinese Lueyang black-bone Silky fowl egg white pp.1532-2297, 2019, https://doi.org/10.1080/10826068.2018.1476887
  2. Lysozymes in Fish vol.69, pp.50, 2021, https://doi.org/10.1021/acs.jafc.1c06676