Abstract
This research was aimed at isolating and identifying the predominant lactic acid bacteria (LAB) in the traditional Chinese salt-fermented soybean food, douchi, from Yunnan, China. The predominant LAB present were isolated and identified by conventional culture-dependent methods combined with molecular biological methods. Two hundred and sixty isolates were obtained from thirty kinds of traditional fermented douchi from six cities and counties in Yunnan, and those strains were divided into twelve groups by their morphological and biochemical characteristics. Based on 16S ribosomal DNA (rDNA) sequencing and analysis, 56 representative strains were identified as belonging to 6 genera and 14 species: Lactobacillus (4 spp.), Weissella (3 spp.), Pediococcus (2 spp.), Staphylococcus (2 spp.), Enterococcus (1 sp.), and Bacillus (2 spp.). The results show that douchi contains a large natural population of LAB of diverse composition from which some strains could be selected as starters for functional fermented foods. This is the first study on the original douchi from Yunnan, and the results suggest that it may be a useful source for the isolation of LAB. This study has also laid a foundation for further research on developing functional douchi products.
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References
Abriouel, H., Martín-Platero, A., Maqueda, M., Valdivia, E., Martínez-Bueno, M., 2008. Biodiversity of the microbial community in a Spanish farmhouse cheese as revealed by culture-dependent and culture-independent methods. Int. J. Food Microbiol., 127(3):200–208. [doi:10.1016/j.ijfoodmicro.2008.07.004]
Benito, M.J., Serradillaa, M.J., Ruiz-Moyano, S., Martín, A., Pérez-Nevado, F., Córdoba, M.G., 2008. Rapid differentiation of lactic acid bacteria from autochthonous fermentation of Iberian dry-fermented sausages. Meat Sci., 80(3):656–661. [doi:10.1016/j.meatsci.2008.03.002]
Briggiler Marcó, M., Mercanti, D., Reinheimer, J.A., Quiberoni, A., 2011. Performance of spontaneous phage-resistant derivatives of Lactobacillus plantarum in fermented milk manufacture. Int. Dairy J., 21(11): 857–862. [doi:10.1016/j.idairyj.2011.05.011]
Chao, S.H., Tomii, Y., Watanabe, K., Tsai, Y.C., 2008. Diversity of lactic acid bacteria in fermented brines used to make stinky tofu. Int. J. Food Microbiol., 123(1–2): 134–141. [doi:10.1016/j.ijfoodmicro.2007.12.010]
Chen, Y.S., Yanagida, F., Shinohara, T., 2005. Isolation and identification of lactic acid bacteria from soil using an enrichment procedure. Lett. Appl. Microbiol., 40(3): 195–200. [doi:10.1111/j.1472-765X.2005.01653.x]
Chen, Y.S., Yanagida, F., Hsu, J.S., 2006. Isolation and characterization of lactic acid bacteria from dochi (fermented black beans), a traditional fermented food in Taiwan. Lett. Appl. Microbiol., 43(2):229–235. [doi:10.1111/j.1472-765X.2006.01922.x]
Chin, H.S., Breidt, F., Fleming, H.P., Shin, W.C., Yoon, S.S., 2006. Identification of predominant bacterial isolates from the fermenting kimchi using ITS-PCR and partial 16S rDNA sequence analyses. J. Microbiol. Biotechnol., 16(1):68–76.
Chun, J., Lee, J.H., Jung, Y., Kim, M., Kim, S., Kim, B.K., Lim, Y.W., 2007. EzTaxon: a web-based tool for the identification of prokaryotes based on 16S ribosomal RNA gene sequences. Int. J. Syst. Evol. Microbiol., 57(10):2259–2261. [doi:10.1099/ijs.0.64915-0]
de Man, J.C., Rogosa, M., Sharpe, M.E., 1960. A medium for the cultivation of lactobacilli. J. Appl. Microbiol., 23(1): 130–135. [doi:10.1111/j.1365-2672.1960.tb00188.x]
Devriese, L.A., Laurier, L., de Herdt, P., Hasbrouck, F., 1992. Enterococcal and Streptococcal species isolated from faeces of calves, young cattle and dairy cows. J. Appl. Microbiol., 72(1):29–31. [doi:10.1111/j.1365-2672.1992. tb04877.x]
Harrigan, W.F., McMance, M.E., 1976. Statistical Methods for the Selection and Examination of Microbial Colonies. In: Harrigan, W.F., Mc Mance, M.E. (Eds.), Laboratory Methods in Food and Dairy Microbiology. Academic Press, London, p.47–49.
Kim, B., Lee, J., Jang, J., Kim, J., Han, H., 2003. Leuconostoc inhae sp. nov., a lactic acid bacterium isolated from kimchi. Int. J. Syst. Evol. Microbiol., 53(4):1123–1126. [doi:10.1099/ijs.0.02463-0]
Kim, T.W., Lee, J.H., Kim, S.E., Park, M.H., Chang, H.C., Kim, H.Y., 2009. Analysis of microbial communities in doenjang, a Korean fermented soybean paste, using nested PCR-denaturing gradient gel electrophoresis. Int. J. Food Microbiol., 131(2–3):265–271. [doi:10.1016/j. ijfoodmicro.2009.03.001]
Konings, W.N., Kok, J., Kuipers, O.P., Poolman, B., 2000. Lactic acid bacteria: the bugs of the new millennium. Curr. Opin. Microbiol., 3(3):276–282. [doi:10.1016/S1369-5274(00)00089-8]
Kubo, Y., Rooney, A.P., Tsukakoshi, Y., Nakagawa, R., Hasegawa, H., Kimura, K., 2011. Phylogenetic analysis of Bacillus subtilis strains applicable to natto (fermented soybean) production. Appl. Environ. Microbiol., 77(18): 6463–6469. [doi:10.1128/AEM.00448-11]
Larkin, M.A., Blackshields, G., Brown, N.P., Chenna, R., McGettigan, P.A., McWilliam, H., Valentin, F., Wallace, I.M., Wilm, A., Lopez, R., et al., 2007. Clustal W and Clustal X version 2. Bioinformatics, 23(21):2947–2948. [doi:10.1093/bioinformatics/btm404]
Lim, S.M., Im, D.S., 2009. Screening and characterization of probiotic lactic acid bacteria isolated from Korean fermented foods. J. Microbiol. Biotechnol., 19(2):178–186. [doi:10.4014/jmb.0804.269]
Liu, P., Shen, S.R., Ruan, H., Zhou, Q., Ma, L.L., He, G.Q., 2011. Production of conjugated linoleic acids by Lactobacillus plantarum strains isolated from naturally fermented Chinese pickles. J. Zhejiang Univ.-Sci. B (Biomed. & Biotechnol.), 12(11):923–930. [doi:10.1631/jzus.B1100072]
Mohd Adnan, A.F., Tan, I.K., 2007. Isolation of lactic acid bacteria from Malaysian foods and assessment of the isolates for industrial potential. Bioresour. Technol., 98(7):1380–1385. [doi:10.1016/j.biortech.2006.05.034]
Moreno, M.R.F., Leisner, J.J., Tee, L.K., Ley, C., Radu, S., Rusul, G., Vancanneyt, M., de Vuyst, L., 2002. Microbial analysis of Malaysian tempeh, and characterization of two bacteriocins produced by isolates of Enterococcus faecium. J. Appl. Microbiol., 92(1):147–157. [doi:10. 1046/j.1365-2672.2002.01509.x]
Ndagano, D., Lamoureux, T., Dortu, C., Vandermoten, S., Thonart, P., 2011. Antifungal activity of 2 lactic acid bacteria of the Weissella genus isolated from food. J. Food Sci., 76(6):M305–M311. [doi:10.1111/j.1750-3841.2011.02257.x]
Nout, M.J.R., Bakshi, D., Sarkar, P.K., 1998. Microbiological safety of kinema, a fermented soybean food. Food Control, 9(6):357–362. [doi:10.1016/S0956-7135(98)00126-1]
Reid, G., 1999. The scientific basis for probiotic strains of Lactobacillus. Appl. Environ. Microbiol., 65(9):3763–3766.
Saitou, N., Nei, M., 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol., 4(4):619–629.
Schmidt, T.M., DeLong, E.F., Pace, N.R., 1991. Analysis of a marine picoplankton community by 16S rRNA gene cloning and sequencing. J. Bacteriol., 173(14):4371–4378.
Tamura, K., Dudley, J., Nei, M., Kumar, S., 2007. MEGA 4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol. Biol. Evol., 24(8):1596–1599. [doi:10.1093/molbev/msm092]
Thompson, J.D., Gibson, T.J., Plewniak, F., Jeanmougin, F., Higgins, D.G., 1997. The Clustal X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res., 25(24): 4876–4882. [doi:10.1093/nar/25.24.4876]
Tsai, Y.H., Kung, H.F., Chang, S.C., Lee, T.M., Wei, C.I., 2007. Histamine formation by histamine-forming bacteria in douche, a Chinese traditional fermented soybean product. Food Chem., 103(4):1305–1311. [doi:10.1016/j. foodchem.2006.10.036]
Wattiau, P., Renard, M.E., Ledent, P., Debois, V., Blackkman, G., Agathos, S.N., 2001. A PCR test to identify Bacillus subtilis and closely related species and its application to the monitoring of waste water treatment. Appl. Microbiol. Biotechnol., 56(5–6):816–819. [doi:10.1007/s002530100691]
Won, T.J., Kim, B., Oh, E.S., Bang, J.S., Lee, Y.J., Yoo, J.S., Yu, H., Yoon, J., Hyung, K.E., Park, S.Y., et al., 2011. Immunomodulatory activity of Lactobacillus strains isolated from fermented vegetables and infant stool. Can. J. Physiol. Pharmacol., 89(6):429–434. [doi:10.1139/y11-047]
Yanagida, F., Chen, Y.S., Shinohara, T., 2005. Isolation and characterization of lactic acid bacteria from soils in vineyards. J. Gen. Appl. Microbiol., 51(5):313–318. [doi:10.2323/jgam.51.313]
Zago, M., Fornasari, M.E., Carminati, D., Burns, P., Suàrez, V., Vinderola, G., Reinheimer, J., Giraffa, G., 2011. Characterization and probiotic potential of Lactobacillus plantarum strains isolated from cheeses. Food Microbiol., 28(5):1033–1040. [doi:10.1016/j.fm.2011.02.009]
Zhang, J.H., Tatsumi, E., Ding, C.H., Li, L.T., 2006. Angiotensin I-converting enzyme inhibitory peptides in douchi, a Chinese traditional fermented soybean product. Food Chem., 98(3):551–557. [doi:10.1016/j.foodchem.2005.06.024]
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Project supported by the State Key Laboratory of Food Science and Technology of Jiangnan University (No. SKLF-KF-200805) and the Yunnan Provincial Science and Technology Department (No. KKSA200926038), China
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Liu, Cj., Gong, Fm., Li, Xr. et al. Natural populations of lactic acid bacteria in douchi from Yunnan Province, China. J. Zhejiang Univ. Sci. B 13, 298–306 (2012). https://doi.org/10.1631/jzus.B1100221
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DOI: https://doi.org/10.1631/jzus.B1100221