Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지
DOI QR코드

DOI QR Code

Characteristic study and isolation of Bacillus subtilis SRCM 101269 for application of cow manure

우분 적용을 위한 Bacillus subtilis SRCM 101269의 분리 및 특성 연구

  • Jeon, SaeBom (Department of Biological Sciences, Chonbuk National University) ;
  • Oh, HyeonHwa (Department of Biological Sciences, Chonbuk National University) ;
  • Uhm, Tai-Boong (Department of Biological Sciences, Chonbuk National University) ;
  • Cho, Jae-Young (Department of Bioenvironmental Chemistry, Chonbuk National University) ;
  • Yang, Hee-Jong (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Jeong, Do-Youn (Microbial Institute for Fermentation Industry (MIFI))
  • 전새봄 (전북대학교 자연과학대학 생물학과) ;
  • 오현화 (전북대학교 자연과학대학 생물학과) ;
  • 엄태붕 (전북대학교 자연과학대학 생물학과) ;
  • 조재영 (전북대학교 농과대학 생물환경화학과) ;
  • 양희종 ((재)발효미생물산업진흥원) ;
  • 정도연 ((재)발효미생물산업진흥원)
  • Received : 2016.01.14
  • Accepted : 2016.02.22
  • Published : 2016.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

Bacillus subtilis SRCM 101269 having safety and amo gene isolated from Korean traditional fermented food and their investigated characterization to apply the cow manure such as cellulase and xylanase activities, 16S rRNA sequencing, and ability of removal of livestock manure odor. Cow manure application results for the removal of livestock manure odor, the ammonia gas was reduced more than two-folder compared to the control group after 6 days, and reduced to less than 10 ppm after 9 days. In the case of cow manure added fowl droppings and other wood-based mixture components, ammonia gas maintained constant after 3 days of fermentation. However, in the case of sample inoculated B. subtilis SRCM 101269, ammonia gas reduced in course of fermentation time, and concentration of hydrogen sulfide also reduced for 65 ppm. Changes of nitrite concentration according to fermentation time no showed different for cow manure, however nitrite concentration in mixed livestock manure increased when compared to control. And then sulfate concentration in cow manure decreased, and no showed different when compared to the initial fermentation. No apparent change of sulfate concentration in mixed livestock manure detected. Through the previously studies, B. subtilis SRCM 101269 has high potential in industrial application manufacturing the cow manure as removal of livestock manure odor.

안전성이 확보되고 amo 유전자가 있는 토종미생물 Bacillus subtilis SRCM 101269는 전통발효식품으로부터 분리하였으며, cellulase, xylanase 생성능, 동정, 악취저감화능 등을 조사하여 우분 적용을 위한 특성 연구를 진행하였다. 가축분뇨 악취제어 관련 우분 적용 결과, 발효 6일 경과 후 대조구에 비해 2배 이상 암모니아 가스가 감소하였고 9일 경과 후 10 ppm 미만으로 농도가 감소하였다. 우분에 계분과 기타 목질성분이 첨가된 혼합분의 경우 발효 3일 후부터 일정 농도로 암모니아 가스가 유지되는데 반해, B. subtilis SRCM 101269를 접종한 시료에서만 발효시간의 경과에 따라 암모니아 가스가 감소하였으며, 황화수소의 농도 또한 9일이 경과한 시료에서 65 ppm으로 감소됨이 확인되었다. 발효 경과에 따른 아질산염과 황산염의 시료 내 변화량을 측정한 결과 우분에서의 아질산염은 변화가 없었으나, 혼합분에서는 대조구에 비교하여 증가하였다. 황산염의 경우 우분에서는 발효초기보다 감소하였으나 대조구와 비교하여 큰 차이를 보이지 않았고, 혼합분에서는 변화가 없는 것으로 분석되었다. 최종적으로 앞선 연구들을 토대로 B. subtilis SRCM 101269 균주의 우분퇴비 생산의 산업적 활용이 가능함을 확인하였다.

Keywords

References

  1. Atkinson, C.F., Jones, D.D., and Gauthire, J.J. 1996. Biodegradabilities and microbial activities during composting of municipal solid waste in bench-scale reactors. Compost Sci. Util. 4, 14-23. https://doi.org/10.1080/1065657X.1996.10701848
  2. Atlas, R.M. and Bartha, R. 1993. Microbial ecology-fundermentals and application, 3th ed., pp. 323-332. The Benjamin/Cummings Publishings Company, Redwood city, California, USA.
  3. Baek, H.J., Zo, Y.G., and Ahn, T.S. 2009. Hydrolytic and metabolic capacities of thermophilic Geobacillus isolated from litter deposit of a lakeshore. Korean J. Microbiol. 1, 32-40.
  4. Chakravorty, S., Helb, D., Burday, M., Connell, N., and Alland, D. 2007. A detailed analysis of 16S ribosomal RNA gene segments for the diagnosis of pathogenic bacteria. J. Microbiol. Methods 69, 330-339. https://doi.org/10.1016/j.mimet.2007.02.005
  5. Hooper, A.B., Vannelli, T., Bergmann, D.J., and Arciero, D.M. 1997. Enzymology of the oxidation of ammonia to nitrite by bacteria. Antonie van Leeuwenhoek 71, 59-67. https://doi.org/10.1023/A:1000133919203
  6. Jang, Y.K., Jung, B.J., Kim, J., Song, K.B., Kim, H.J., and Yoo, Y.H. 2009. Assessment of odor characterization and odor unit from livestock facilities by animals. J. Environ. Impact Assessment 19, 29-38.
  7. Kappler, U., Friedrich, C.G., Truper, H.G., and Dahl, C. 2001. Evidence for two pathways of thiosulfate oxidation in Starkeya novella (formerly Thiobacillus novellus). Arch. Microbiol. 175, 102-111. https://doi.org/10.1007/s002030000241
  8. Kim, Y.S., Kang, M.K., Bae, K.S., Lee, K.S., and Rhee, Y.H. 1997. Changes in physic-chemical and microbiological parameters during active composting of cattle manure. Korean J. Microbiol. 33, 267-273.
  9. Kim, S.Y., Kim, H., and Chae, H.J. 2003. Isolation and characterization of microorganisms for the development of fermentation accelerator of animal manure. Korean J. Biotechnol. Bioeng. 18, 466-472.
  10. Kim, K.D., Kim, K.Y., and Hahm, Y.T. 2004. Studies on the isolation and identification of Bacillus sp. for the composting of swine manure and the removal of malodorous gases from its liquid compost. Korean J. Microbiol. 40, 154-159.
  11. Kim, N.J., Mitsuyo, M., and Shoda, M. 2000. Comparison of organic and inorganic packing materials in the removal of ammonia gas in biofilters. J. Hazard. Mater. 72, 77-90. https://doi.org/10.1016/S0304-3894(99)00160-0
  12. Kim, H.S., Park, S.J., Jheong, W.H., Srinivasan, S., and Lee, S.S. 2013. The study on the effect of efficient microorganism for early stabilization of the burial sites. Korean J. Microbiol. 49, 343-352. https://doi.org/10.7845/kjm.2013.3043
  13. Kolmert, A., Wikstrom, P., and Hallberg, K.B. 2000. A fast and simple turbidimetric method for the determination of sulfate in sulfate-reducing bacterial cultures. J. Microbiol. Methods 41, 179-184. https://doi.org/10.1016/S0167-7012(00)00154-8
  14. Koops, H.P., Bottcher, B., Moller, U.C., Pommerening-Roser, A., and Steh, G. 1991. Classification of eight new species of ammoniaoxidizing bacteria: Nitrosomonas communis sp. nov., Nitrosomonas urea sp. nov., Nitrosomonas aestuarii sp. nov., Nitrosomonas marina sp. nov., Nitrosomonas nitrosa sp. nov., Nitrosomonas eutropha sp. nov., Nitrosomonas oligotropha sp. nov. and Nitrosomonas halophila sp. nov. J. Gen. Microbiol. 137, 1689-1699. https://doi.org/10.1099/00221287-137-7-1689
  15. Kuroda, K., Hanajima, D., Fukumoto, Y., Suzuki, K., Kawamoto, S., Shima, J., and Haga, K. 2004. Isolation of thermophilic ammonium-tolerant bacterium and its application to reduce ammonia emission during composting of animal wastes. Biosci. Biotechnol. Biochem. 68, 286-292. https://doi.org/10.1271/bbb.68.286
  16. Lee, S.J. and Lee, E.Y. 2009. Screening and isolation of ammonia removal microorganism for the improvement of livestock environment. Korean J. Microbiol. Biotechnol. 37, 408-412.
  17. Lim, J.S., Jo, W.S., and Lee, E.Y. 2006. Removal of mixed gases of $H_2S$ and $NH_3$ by the biofilter packed with biomedia. Clean Technol. 12, 165-170.
  18. Matulewich, V.A. and Finstein, M.S. 1978. Distribution of autotrophic nitrifying bacteria in a polluted river (the Passaic). Appl. Environ. Microbiol. 35, 61-71.
  19. Ministry of Environment. 2014. Odor prevention act enforcement rules. Ministry of Knowledge Economy, Korea.
  20. Nakamura, K., Miki, H., and Amano, Y. 1990. Cell growth and accumulation of Thiobacillus thiooxidans S3 in a pH-controlled thiosulfate medium. J. Gen. Appl. Microbiol. 36, 369-376. https://doi.org/10.2323/jgam.36.369
  21. Nam, B.S., Ryu, W.R., Lee, Y.H., Kim, J.M., and Cho, M.H. 1999. Isolation and characterization of ammonia and nitrite nitrogen oxidizing strains. Korean J. Biotechnol. Bioeng. 14, 76-81.
  22. Oh, Y.S., Kim, K.H., Koo, Y.S., Kim, M.S., Seol, M.H., Chon, S.S., Choi, Y.J., and Ahn, D.H. 2006. Odor emission characteristics in livestock waste treatment facilities. Korean J. Odor Res. Eng. 5, 1-9.
  23. Papadimitrious, E.K. and Balis, C. 1996. Comparative study of parameters to evaluate and monitor the rate of a composting process. Compost Sci. Util. 4, 52-61. https://doi.org/10.1080/1065657X.1996.10701853
  24. Park, S., Jung, D.H., Yoo, E.S., and Kim, M.I. 2009. Evaluation of complex odor and odorous compounds in a pilot-scale ultra thermophilic aerobic composting process. J. Korean Geoenvironmental Society 10, 33-39.
  25. Ryckeboer, J., Mergaert, J., Coosemans, J., Deprins, K., and Swings, J. 2003. Microbiological aspects of biowaste during composting in a monitored compost bin. J. Appl. Microbiol. 94, 127-137. https://doi.org/10.1046/j.1365-2672.2003.01800.x
  26. Schutz, M., Maldener, I., Griesbeck, C., and Hauska, G. 1999. Sulfide-quinone reductase from Rhodobacter capsulatus: requirement for growth, periplasmic localization, and extension of gene sequence analysis. J. Bacteriol. 181, 6516-6523.
  27. Shin, H.J. 2002. Effect on livestock manure composting by the enriched microbial population. Kor. J. Life Science 12, 129-135. https://doi.org/10.5352/JLS.2002.12.2.129
  28. Stutzenberger, F.J. 1971. Cellulase production by Thermomonospora curvata isolated from municipal solid waste compost. Appl. Microbiol. 22, 147-152.
  29. Tamura, K. and Nei, M. 1993. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Mol. Biol. Evol. 10, 512-526.
  30. Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., and Kumar, S. 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
  31. Thompson, J.D., Higgins, D.G., and Gibson, T.J. 1994. Clustal W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22, 4673-4680. https://doi.org/10.1093/nar/22.22.4673
  32. Updegraff, D.M. 1969. Semimicro determination of cellulose in biological materials. Anal. Biochem. 32, 420-424. https://doi.org/10.1016/S0003-2697(69)80009-6
  33. Ushida, K., Hashizume, K., Miyazaki, K., Kojima, Y., and Takakuwa, S. 2003. Isolation of Bacillus sp. as a volatile sulfur-degrading bacterium and its application to reduce the fecal odor of pig. Asian-Australas. J. Anim. Sci. 16, 1795-1798. https://doi.org/10.5713/ajas.2003.1795
  34. Varel, V.H. 2002. Livestock manure odor abatement with plant-derived oils and nitrogen conservation with urease inhibitors: A review. J. Anim. Sci. 80, E1-E7. https://doi.org/10.2527/2002.8011
  35. Vermelho, A.B., Meirelles, M.N.L., Lopes, A., Petinate, S.D.G., Chaia, A.A., and Branquinha, M.H. 1996. Detection of extracellular proteases from microorganisms on agar plates. Mem. Inst. Oswaldo Cruz. 91, 755-760. https://doi.org/10.1590/S0074-02761996000600020
  36. Yoon, Y.M., An, G.H., Kim, J.K., Ahn, S.H., Cha, Y.L., Yang, J.W., Yu, K.D., Ahn, J.W., Moon, Y.H., Koo, B.C., et al. 2014. Xylanase activity of Bacillus pumilus H10-1 isolated from Ceratotherium simum feces. Korean Soc. Biotechnol. Bioeng. J. 5, 316-322.