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Isolation and characterization of cellulolytic yeast belonging to Moesziomyces sp. from the gut of Grasshopper

메뚜기의 내장에서 분리한 Moesziomyces 속에 속하는 셀룰로오스 분해 효모의 분리 및 특성

  • Kim, Ju-Young (Department of Bio & Environmental Technology, College of Natural Science, Seoul Women's University) ;
  • Jung, Hee-Young (College of Agricultural and Life Sciences, Kyungpook National University) ;
  • Park, Jong-Seok (School of Biological Sciences, College of Natural Sciences, Chungbuk National University) ;
  • Cho, Sung-Jin (School of Biological Sciences, College of Natural Sciences, Chungbuk National University) ;
  • Lee, Hoon Bok (Department of Bio & Environmental Technology, College of Natural Science, Seoul Women's University) ;
  • Sung, Gi-Ho (Institute for Healthcare and Life Science, International St. Mary's Hospital and College of Medicine, Catholic Kwandong University) ;
  • Subramani, Gayathri (Department of Bio & Environmental Technology, College of Natural Science, Seoul Women's University) ;
  • Kim, Myung Kyum (Department of Bio & Environmental Technology, College of Natural Science, Seoul Women's University)
  • 김주영 (서울여자대학교 자연과학대학) ;
  • 정희영 (경북대학교 농업생명과학대학) ;
  • 박종석 (충북대학교 자연과학대학 생물과학부) ;
  • 조성진 (충북대학교 자연과학대학 생물과학부) ;
  • 이훈복 (서울여자대학교 자연과학대학) ;
  • 성기호 (가톨릭관동대학교 국제성모병원 및 의과대학) ;
  • 가야쓰리 수브라마니 (서울여자대학교 자연과학대학) ;
  • 김명겸 (서울여자대학교 자연과학대학)
  • Received : 2019.06.10
  • Accepted : 2019.09.04
  • Published : 2019.09.30

Abstract

An intensive interaction between yeasts and insects has highlighted their relevance for attraction to food and for the insect's development and behavior. Yeast associated in the gut of insects secretes cellulase which aided in the food digestion (cellulose degradation). Three strains of cellulose-degrading yeast were isolated from the gut of adult grasshoppers collected in Gyeonggi Province, South Korea. The strains $ON22^T$, $G10^T$, and $G15^T$, showed positive cellulolytic activity in the carboxymethyl cellulose (CMC)-plate assay. The phylogenetic tree based on sequence analysis of D1/D2 domains of the large subunit rRNA gene and the internal transcribed spacer (ITS) regions revealed that the strains $ON22^T$ (100 and 98.4% sequence similarities in D1/D2 domains and ITS) and $G10^T$ (99.8 and 99.5% in D1/D2 domain and ITS region) were most closely related to the species Moesziomyces aphidis JCM $10318^T$; $G15^T$ (100% in D1/D2 domains and ITS) belongs to the species Moesziomyces antarcticus JCM $10317^T$, respectively. Morphology and biochemical test results are provided in the species description. Cellulase with its massive applicability has been used in various industrial processes such as biofuels like bioethanol productions. Therefore, this is the first report of the cellulolytic yeast strains $ON22^T$, $G10^T$, and $G15^T$ related to the genus Moesziomyces in the family Ustilaginaceae (Ustilaginales), in Korea.

Keywords

Moesziomyces sp.;D1/D2 domain;grasshopper;ITS;rRNA;yeast

Acknowledgement

Supported by : National Institute of Biological Resources (NIBR)

References

  1. Boekhout T and Fell JW. 1998. Pseudozyma Bandoni emend. Boekhout and a comparison with the yeast state of Ustilago maydis (de Candolle) Corda. In Kurtzman, C.P. and Fell, J.W. (eds.), The Yeasts: a Taxonomic Study. Elsevier Science Publish, Amsterdam, pp. 790-797.
  2. Cubero F, Crespo A, Fatehi J, and Bridge DP. 1998. DNA extraction and PCR amplification method suitable for fresh, herbariumstored, lichenized, and other fungi. Plant Syst. Evol. 216, 243-249.
  3. Dashtban M, Maki M, Leung KT, Mao C, and Qin W. 2010. Cellulase activities in biomass conversion: Measurement methods and comparison. Crit. Rev. Biotechnol. 30, 302-309. https://doi.org/10.3109/07388551.2010.490938
  4. Dillon RJ and Dillon VM. 2004. The gut bacteria of insects: nonpathogenic interactions. Annu. Rev. Entomol. 49, 71-92. https://doi.org/10.1146/annurev.ento.49.061802.123416
  5. Felsenstein J. 1985. Confidence limits on phylogenies: An approach using the bootstrap. Evolution 39, 783-791. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x
  6. Fitch MW. 1971. Toward defining the course of evolution: minimum change for a specific tree topology. Syst. Zool. 20, 406-416. https://doi.org/10.2307/2412116
  7. Fujita Y, Takahashi S, Ueda M, Tanaka A, Okada H, Morikawa Y, Kawaguchi T, Arai M, Fukuda H, and Kondo A. 2002. Direct and efficient production of ethanol from cellulosic material with a yeast strain displaying cellulolytic enzymes. Appl. Environ. Microbiol. 68, 5136-5141. https://doi.org/10.1128/AEM.68.10.5136-5141.2002
  8. Goto S, Sugiyama J, and Iizuka H. 1969. A taxonomic study of Antarctic yeasts. Mycologia 61, 748-774. https://doi.org/10.1080/00275514.1969.12018794
  9. Hall TA. 1999. BioEdit: A user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp. Ser. 41, 95-98.
  10. Ito J, Fujita Y, Ueda M, Fukuda H, and Kondo A. 2004. Improvement of cellulose-degrading ability of a yeast strain displaying Trichoderma reesei endoglucanase II by recombination of cellulose-binding domains. Biotechnol. Prog. 20, 688-691. https://doi.org/10.1021/bp034332u
  11. Johnsen HR and Krause K. 2014. Cellulase activity screening using pure carboxymethylcellulose: application to soluble cellulolytic samples and to plant tissue prints. Int. J. Mol. Sci. 15, 830-838. https://doi.org/10.3390/ijms15010830
  12. Kumar S, Stecher G, and Tamura K. 2016. MEGA7: Molecular Evolutionary Genetics Analysis Version 7.0 for Bigger Datasets. Mol. Biol. Evol. 33, 1870-1874. https://doi.org/10.1093/molbev/msw054
  13. Kurtzman CP, Fell JW, Boekhout T, and Robert V. 2011. Chapter 7 - Methods for Isolation, Phenotypic Characterization and Maintenance of Yeasts. In Kurtzman CP, Fell JW, and Boekhout T (eds.). The Yeasts (Fifth Edition), pp. 87-110. Elsevier, London, UK.
  14. Kurtzman CP and Robnett CJ. 1998. Identification and phylogeny of ascomycetous yeasts from analysis of nuclear large subunit (26S) ribosomal DNA partial sequences. Antonie van Leeuwenhoek 73, 331-371. https://doi.org/10.1023/A:1001761008817
  15. Lee SJ, Kim SR, Yoon HJ, Kim I, Lee KS, Je YH, Lee SM, Seo SJ, Dae Sohn H, and Jin BR. 2004. cDNA cloning, expression, and enzymatic activity of a cellulase from the mulberry longicorn beetle, Apriona germari. Comp. Biochem. Physiol. B Biochem. Mol. Biol. 139, 107-116. https://doi.org/10.1016/j.cbpc.2004.06.015
  16. Parahym AM, da Silva CM, Domingos Ide F, Goncalves SS, Rodrigues Mde M, de Morais VL, and Neves RP. 2013. Pulmonary infection due to Pseudozyma aphidis in a patient with burkitt lymphoma: first case report. Diagn. Microbiol. Infect. Dis. 75, 104-106. https://doi.org/10.1016/j.diagmicrobio.2012.09.010
  17. Romao-Dumaresq AS, Dourado MN, Favaro LC, Mendes R, Ferreira A, and Araujo WL. 2016. Diversity of cultivated fungi associated with conventional and transgenic sugarcane and the interaction between endophytic Trichoderma virens and the host plant. PLoS One 11, e0158974. https://doi.org/10.1371/journal.pone.0158974
  18. Saitou N and Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406-425.
  19. Sugita T, Nishikawa A, Ikeda R, and Shinoda T. 1999. Identification of medically relevant Trichosporon species based on sequences of internal transcribed spacer regions and construction of a database for Trichosporon identification. J. Clin. Microbiol. 37, 1985-1993.
  20. Takahashi S, Ueda M, and Tanaka A. 2000. Effect of the truncation of the C-terminal region of Kex2 endoprotease on processing of the recombinant Rhizopus oryzae lipase precursor in the coexpression system in yeast. J. Mol. Catal. B Enzym. 10, 233-240. https://doi.org/10.1016/S1381-1177(00)00130-2
  21. Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, and Higgins DG. 1997. The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res. 25, 4876-4882. https://doi.org/10.1093/nar/25.24.4876
  22. Tomme P, Warren RAJ, and Gilkes NR. 1995. Cellulose hydrolysis by bacteria and fungi. Adv. Microbiol. Physiol. 37, 1-81.
  23. Trevan, M. 1987. Techniques of Immobilization. In Immobilized Enzymes. An Introduction and Applications in Biotechnology (Trevan, M., ed.), pp. 1-9, Wiley, Chichester-New York.
  24. Van Rensburg P, Van Zyl WH, and Pretorius IS. 1998. Engineering yeast for efficient cellulose degradation. Yeast 14, 67-76. https://doi.org/10.1002/(SICI)1097-0061(19980115)14:1<67::AID-YEA200>3.0.CO;2-T
  25. Vu D, Groenewald M, Szöke S, Cardinali G, Eberhardt U, Stielow B, de Vries M, Verkleij GJM, Crous PW, Boekhout T, et al. 2016. DNA barcoding analysis of more than 9000 yeast isolates contributes to quantitative thresholds for yeast species and genera delimitation. Stud. Mycol. 85, 91-105. https://doi.org/10.1016/j.simyco.2016.11.007
  26. Wang QM, Begerow D, Groenewald M, Liu XZ, Theelen B, Bai FY, and Boekhout T. 2015. Multigene phylogeny and taxonomic revision of yeasts and related fungi in the Ustilaginomycotina. Stud. Mycol. 81, 55-83. https://doi.org/10.1016/j.simyco.2015.10.004
  27. Wang QM, Jia JH, and Bai FY. 2006. Pseudozyma hubeiensis sp. nov. and Pseudozyma shanxiensis sp. nov., novel ustilaginomycetous anamorphic yeast species from plant leaves. Int. J. Syst. Evol. Microbiol. 56, 289-293. https://doi.org/10.1099/ijs.0.63827-0
  28. Wei YH, Lee FL, Hsu WH, Chen WH, Chen CC, Wen CY, Lin SJ, Chu WS, Yuan GF, and Liou GY. 2005. Pseudozyma antarctica in Taiwan: a description based on morphological, physiological and molecular characteristics. Bot. Bull. Acad. Sin. 46, 223-229.
  29. White T, Bruns T, Lee S, and Taylor J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In Innis M, Gelfand D, Shinsky J, and White T (eds.). PCR Protocols: A Guide to Methods and Applications, pp. 315-322. Academic Press.
  30. Yeoh HH, Khew E, and Lim G. 1985. A simple method for screening cellulolytic fungi. Mycologia 77, 161-162. https://doi.org/10.1080/00275514.1985.12025077
  31. Zhang YH, Hong J, and Ye X. 2009. Cellulase assays. Methods Mol. Biol. 581, 213-231.
  32. Kimura M. 1980. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J. Mol. Evol. 16, 111-120. https://doi.org/10.1007/BF01731581