Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Identification of Lactic Acid Bacteria in Galchi- and Myeolchi-Jeotgal by 16S rRNA Gene Sequencing, MALDI-TOF Mass Spectrometry, and PCR-DGGE

  • Lee, Yoonju (Institute of Life Sciences & Resources and Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Cho, Youngjae (Institute of Life Sciences & Resources and Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Kim, Eiseul (Institute of Life Sciences & Resources and Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Kim, Hyun-Joong (Institute of Life Sciences & Resources and Department of Food Science and Biotechnology, Kyung Hee University) ;
  • Kim, Hae-Yeong (Institute of Life Sciences & Resources and Department of Food Science and Biotechnology, Kyung Hee University)
  • Received : 2018.03.27
  • Accepted : 2018.06.04
  • Published : 2018.07.28
Download PDF
⟨ Previous Next ⟩

Abstract

Jeotgal is a Korean traditional fermented seafood with a high concentration of salt. In this study, we isolated lactic acid bacteria (LAB) from galchi (Trichiurus lepturus, hairtail) and myeolchi (Engraulis japonicas, anchovy) jeotgal on MRS agar and MRS agar containing 5% NaCl (MRS agar+5% NaCl), and identified them by using 16S rRNA gene sequencing and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as culture-dependent methods. We also performed polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) as a culture-independent method to identify bacterial communities. Five samples of galchi-jeotgal and seven samples of myeolchi-jeotgal were collected from different regions in Korea. A total of 327 and 395 colonies were isolated from the galchi- and myeolchi-jeotgal samples, respectively. 16S rRNA gene sequencing and MALDI-TOF MS revealed that the genus Pediococcus was predominant on MRS agar, and Tetragenococcus halophilus on MRS agar+5% NaCl. PCR-DGGE revealed that T. halophilus, Tetragenococcus muriaticus, and Lactobacillus sakei were predominant in both types of jeotgal. T. halophilus was detected in all samples. Even though the same species were identified by both culture-dependent and -independent methods, many species identified by the culture-dependent methods were not in the bacterial list identified by the culture-independent methods. The distribution of bacteria in galchi-jeotgal was more diverse than in myeolchi-jeotgal. The diverse LAB in galchi- and myeolchi-jeotgals can be further studied as candidates for starter cultures to produce fermented foods.

Keywords

References

  1. Koo O, Lee S, Chung K, Jang D, Yang H. 2016. Korean traditional fermented fish products: jeotgal. J. Ethn. Foods 3: 107-116. https://doi.org/10.1016/j.jef.2016.06.004
  2. Marui J, Boulom S, Panthavee W, Momma M, Kusumoto K, Nakahara K, et al. 2015. Culture-independent bacterial community analysis of the salty-fermented fish paste products of Thailand and Laos. Biosci. Microbiota Food Health 34: 45-52. https://doi.org/10.12938/bmfh.2014-018
  3. Choi DK. 2012. The appearance of salted seafood in the east Asia and Vietnam's nuoc mam - focusing on the origin and popularization of fermented sea foods in Korea and in Vietnam. Asian Comp. Folklore 48: 201-246.
  4. Guan L, Cho KH, Lee JH. 2011. Analysis of the cultivable bacterial community in jeotgal, a Korean salted and fermented seafood, and identification of its dominant bacteria. Food Microbiol. 28: 101-113.
  5. Ahmadsah LSF, Kim E, Jung YS, Kim HY. 2018. Identification by PCR-DGGE, SDS-PAGE, and MALDI-TOF MS of LAB and fungi in laru, a fermented starter. J. Microbiol. Biotechnol. 28: 32-39.
  6. Cho Y, Kim E, Lee Y, Han SK, Choo DW, Kim YR, et al. 2017. Rapid and accurate identification of Pediococcus species isolated from Korean fermented foods by MALDITOF MS with local database extension. Int. J. Syst. Evol. Microbiol. 67: 744-752. https://doi.org/10.1099/ijsem.0.001626
  7. Kim E, Cho Y, Lee Y, Han SK, Kim CG, Choo DW, et al. 2017. A proteomic approach for rapid identification of Weissella species isolated from Korean fermented foods on MALDI-TOF MS supplemented with an in-house database. Int. J. Food Microbiol. 243: 9-15. https://doi.org/10.1016/j.ijfoodmicro.2016.11.027
  8. Kim MS, Park EJ. 2014. Bacterial communities of traditional salted and fermented seafoods from Jeju Island of Korea using 16S rRNA gene clone library analysis. J. Food Sci. 79: M927-M934. https://doi.org/10.1111/1750-3841.12431
  9. Han S, Hong Y, Kwak H, Kim E, Kim M, Shrivastav A, et al. 2014. Identification of lactic acid bacteria in pork meat and pork meat products using SDS-PAGE, 16S rRNA gene sequencing and MALDI-TOF mass spectrometry. J. Food Saf. 34: 224-232. https://doi.org/10.1111/jfs.12117
  10. Nguyen DT, Van Hoorde K, Cnockaert M, De Brandt E, Aerts M, Binh Thanh L, et al. 2013. A description of the lactic acid bacteria microbiota associated with the production of traditional fermented vegetables in Vietnam. Int. J. Food Microbiol. 163: 19-27. https://doi.org/10.1016/j.ijfoodmicro.2013.01.024
  11. Doan NT, Van Hoorde K, Cnockaert M, De Brandt E, Aerts M, Le Thanh B, et al. 2012. Validation of MALDI-TOF MS for rapid classification and identification of lactic acid bacteria, with a focus on isolates from traditional fermented foods in Northern Vietnam. Lett. Appl. Microbiol. 55: 265-273. https://doi.org/10.1111/j.1472-765X.2012.03287.x
  12. Hong Y, Li J, Qin P, Lee SY, Kim HY. 2015. Predominant lactic acid bacteria in mukeunji, a long-term-aged kimchi, for different aging periods. Food Sci. Biotechnol. 24: 545-550. https://doi.org/10.1007/s10068-015-0071-6
  13. Angelakis E, Million M, Henry M, Raoult D. 2011. Rapid and accurate bacterial identification in probiotics and yoghurts by MALDI-TOF mass spectrometry. J. Food Sci. 76: M568-M572. https://doi.org/10.1111/j.1750-3841.2011.02369.x
  14. Lee SH, Jung JY, Jeon CO. 2014. Effects of temperature on microbial succession and metabolite change during saeu-jeot fermentation. Food Microbiol. 38: 16-25. https://doi.org/10.1016/j.fm.2013.08.004
  15. Hong Y, Yang HS, Li J, Han SK, Chang HC, Kim HY. 2014. Identification of lactic acid bacteria in salted Chinese cabbage by SDS-PAGE and PCR-DGGE. J. Sci. Food Agric. 94: 296-300. https://doi.org/10.1002/jsfa.6257
  16. Han KI, Kim YH, Hwang SG, Jung EG, Patnaik BB, Han YS, et al. 2014. Bacterial community dynamics of salted and fermented shrimp based on denaturing gradient gel electrophoresis. J. Food Sci. 79: M2516-M2522. https://doi.org/10.1111/1750-3841.12707
  17. Roh SW, Kim KH, Nam YD, Chang HW, Park EJ, Bae JW. 2010. Investigation of archaeal and bacterial diversity in fermented seafood using barcoded pyrosequencing. ISME J. 4: 1-16. https://doi.org/10.1038/ismej.2009.83
  18. Kim TW, Lee JH, Park MH, Kim HY. 2010. Analysis of bacterial and fungal communities in Japanese- and Chinesefermented soybean pastes using nested PCR-DGGE. Curr. Microbiol. 60: 315-320. https://doi.org/10.1007/s00284-009-9542-4
  19. Udomsil N, Rodtong S, Choi YJ, Hua Y, Yongsawatdigul J. 2011. Use of Tetragenococcus halophilus as a starter culture for flavor improvement in fish sauce fermentation. J. Agric. Food Chem. 59: 8401-8408. https://doi.org/10.1021/jf201953v
  20. Collins MD, Williams AM, Wallbanks S. 1990. The phylogeny of Aerococcus and Pediococcus as determined by 16S rRNA sequence analysis: description of Tetragenococcus gen. nov. FEMS Microbiol. Lett. 58: 255-262.
  21. Masuda S, Yamaguchi H, Kurokawa T, Shirakami T, Tsuji RF, Nishimura I. 2008. Immunomodulatory effect of halophilic lactic acid bacterium Tetragenococcus halophilus Th221 from soy sauce moromi grown in high-salt medium. Int. J. Food Microbiol. 121: 245-252. https://doi.org/10.1016/j.ijfoodmicro.2007.10.011
  22. Lee H, Choi Y, Hwang I, Hong S, Lee M. 2016. Relationship between chemical characteristics and bacterial community of a Korean salted-fermented anchovy sauce, Myeolchi-Aekjeot. LWT Food Sci. Technol. 73: 251-258. https://doi.org/10.1016/j.lwt.2016.06.007
  23. Jung JY, Lee SH, Jeon CO. 2014. Kimchi microflora: history, current status, and perspectives for industrial kimchi production. Appl. Microbiol. Biotechnol. 98: 2385-2393. https://doi.org/10.1007/s00253-014-5513-1
  24. Kim M, Chun J. 2005. Bacterial community structure in kimchi, a Korean fermented vegetable food, as revealed by 16S rRNA gene analysis. Int. J. Food Microbiol. 103: 91-96. https://doi.org/10.1016/j.ijfoodmicro.2004.11.030
  25. Lee JS, Heo GY, Lee JW, Oh YJ, Park JA, Park YH, et al. 2005. Analysis of kimchi microflora using denaturing gradient gel electrophoresis. Int. J. Food Microbiol. 102: 143-150. https://doi.org/10.1016/j.ijfoodmicro.2004.12.010
  26. Park JM, Shin JH, Lee DW, Song JC, Suh HJ, Chang UJ, et al. 2010. Identification of the lactic acid bacteria in kimchi according to initial and over-ripened fermentation using PCR and 16S rRNA gene sequence analysis. Food Sci. Biotechnol. 19: 541-546. https://doi.org/10.1007/s10068-010-0075-1
  27. Cho J, Lee D, Yang C, Jeon J, Kim J, Han H. 2006. Microbial population dynamics of kimchi, a fermented cabbage product. FEMS Microbiol. Lett. 257: 262-267. https://doi.org/10.1111/j.1574-6968.2006.00186.x
  28. Zheng Y, Zheng XW, Han BZ, Han JS, Nout MJR, Chen JY. 2013. Monitoring the ecology of Bacillus during Daqu incubation, a fermentation starter, using culture-dependent and culture-independent methods. J. Microbiol. Biotechnol. 23: 614-622. https://doi.org/10.4014/jmb.1211.11065
  29. Yan YZ, Qian YL, Ji FD, Chen JY, Han BZ. 2013. Microbial composition during Chinese soy sauce koji-making based on culture dependent and independent methods. Food Microbiol. 34: 189-195. https://doi.org/10.1016/j.fm.2012.12.009

Cited by

  1. Rapid Identification of Staphylococcus Species Isolated from Food Samples by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry vol.29, pp.4, 2019, https://doi.org/10.4014/jmb.1901.01046