Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
권호 표지
DOI QR코드

DOI QR Code

Optimization of Medium Components using Response Surface Methodology for Cost-effective Mannitol Production by Leuconostoc mesenteroides SRCM201425

반응표면분석법을 이용한 Leuconostoc mesenteroides SRCM201425의 만니톨 생산배지 최적화

  • Ha, Gwangsu (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Shin, Su-Jin (Korean culture center of microorganisms (KCCM)) ;
  • Jeong, Seong-Yeop (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Yang, HoYeon (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Im, Sua (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Heo, JuHee (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Yang, Hee-Jong (Microbial Institute for Fermentation Industry (MIFI)) ;
  • Jeong, Do-Youn (Microbial Institute for Fermentation Industry (MIFI))
  • 하광수 ((재)발효미생물산업진흥원) ;
  • 신수진 ((사)한국미생물보존센터) ;
  • 정성엽 ((재)발효미생물산업진흥원) ;
  • 양호연 ((재)발효미생물산업진흥원) ;
  • 임수아 ((재)발효미생물산업진흥원) ;
  • 허주희 ((재)발효미생물산업진흥원) ;
  • 양희종 ((재)발효미생물산업진흥원) ;
  • 정도연 ((재)발효미생물산업진흥원)
  • Received : 2019.07.15
  • Accepted : 2019.08.08
  • Published : 2019.08.30
Download PDF
⟨ Previous Next ⟩

Abstract

This study was undertaken to establish optimum medium compositions for cost-effective mannitol production by Leuconostoc mesenteroides SRCM201425 isolated from kimchi. L. mesenteroides SRCM21425 from kimchi was selected for efficient mannitol production based on fructose analysis and identified by its 16S rRNA gene sequence, as well as by carbohydrate fermentation pattern analysis. To enhance mannitol production by L. mesenteroides SRCM201425, the effects of carbon, nitrogen, and mineral sources on mannitol production were first determined using Plackett-Burman design (PBD). The effects of 11 variables on mannitol production were investigated of which three variables, fructose, sucrose, and peptone, were selected. In the second step, each concentration of fructose, sucrose, and peptone was optimized using a central composite design (CCD) and response surface analysis. The predicted concentrations of fructose, sucrose, and peptone were 38.68 g/l, 30 g/l, and 39.67 g/l, respectively. The mathematical response model was reliable, with a coefficient of determination of $R^2=0.9185$. Mannitol production increased 20-fold as compared with the MRS medium, corresponding to a mannitol yield 97.46% when compared to MRS supplemented with 100 g/l of fructose in flask system. Furthermore, the production in the optimized medium was cost-effective. The findings of this study can be expected to be useful in biological production for catalytic hydrogenation causing byproduct and additional production costs.

본 연구에서는 경제성 있는 생물학적 만니톨 고생산을 위해 선별균주의 만니톨 생산 최적화 배지조성을 RSM 방법을 이용하여 확립하고자 하였다. 먼저 김치로부터 분리된 10균주의 만니톨 생산량과 과당으로부터 만니톨 전환율 분석을 통하여 SRCM201425 균주를 선발하였으며, 선발균주는 16S rRNA 유전자 염기서열과 당 발효 분석을 통하여 Leuconostoc mesenteroides로 동정하였다. Plackett-Burman design (PBD)을 이용하여 만니톨 생산에 영향을 주는 배지 인자를 선별하기 위해 총 11개의 탄소원, 질소원, 무기원소의 영향을 조사하였으며, 통계학적 분석을 통하여 최종적으로 fructose와 sucrose, peptone을 선정하였다. 만니톨 생산을 위한 선별된 각 변수의 최적 농도를 결정하기 위한 방법으로 central composite design (CCD)과 반응표면 분석법을 이용하였으며, 최종적으로 CCD를 통해 만니톨 생산을 위한 배지 조성의 최적 농도는 fructose 38.68 g/l, sucrose 30 g/l, peptone 39.67 g/l으로 예측되었으며, 통계학적 분석을 통해 실험모델의 적합성을 확인하였다. 최종적으로 MRS 배지에서의 생산량의 약 20배의 만니톨을 생산할 수 있었으며, 100 g/l의 fructose가 포함된 MRS 배지 대비 97.46%의 만니톨을 생산하면서, 산업화시 기존 배지 대비 생산비용을 크게 절감할 수 있을 것으로 예상되었다. 본 연구를 통하여 만니톨 생산을 위한 배지 조성의 최적화를 확립하였으며, 만니톨을 생산하기 위한 방법으로 주로 사용되고 있는 고비용의 촉매환원 방법을 대체할 방법을 제시할 수 있는 기초자료로 활용될 수 있을 것으로 기대된다.

Keywords

References

  1. Andreevskaya, M., Johansson, P., Jaaskelainen, E., Ramo, T., Ritari, J., Paulin, L., Bjorkroth, J. and Auvinen, P. 2016. Lactobacillus oligofermentans glucose, ribose and xylose transciptomes show higher similarity between glucose and xylose catabolism-induced responses in the early exponential growth phase. BMC Genomics 17, 539. https://doi.org/10.1186/s12864-016-2840-x
  2. Bong, K. M., Kim, K. M., Seo, M. K., Han, J. H., Park, I. C., Lee, C. W. and Kim, P. I. 2017. Optimization of medium for the carotenoid production by Rhodobacter sphaeroides PS-24 using response surface methodology. Kor. J. Org. Agric. 25, 135-148. https://doi.org/10.11625/KJOA.2017.25.1.135
  3. Ghoreishi, S. M. and Gholami, R. 2009. Innovative strategies for engineering mannitol production. Trends Food Sci. Technol. 20, 263-270. https://doi.org/10.1016/j.tifs.2009.03.006
  4. Grobben, G. J., Peters, S. W. P., Wisselink, H., Weusthuis, R. A., Hoefnagel, M. H. N., Hugenholtz, J. and Eggink, G. 2001. Spontaneous formation of a mannitol-producing variant of Leuconostoc pseudomesenteroides grown in the presence of fructose. Appl. Environ. Microbiol. 67, 2867-2870. https://doi.org/10.1128/AEM.67.6.2867-2870.2001
  5. Ha, G. S., Yang, H. J., Jeong, S. J., Ryu, M. S., Kim, J. W., Yang, H. Y., Shin, S. J., Im, S. A., Seo, J. W., Jeong, S. Y. and Jeong, D. Y. 2018. Medium optimization for enhanced growth of Bacillus subtilis SRCM102046 possessing antibacterial activity using response-surface methodology. Kor. J. Food Preserv. 25, 613-624. https://doi.org/10.11002/kjfp.2018.25.5.613
  6. Jeong, E. J., Moon, D. W., Oh, J. S., Moon, J. S., Seong, H. B., Kim, K. Y. and Han, N. S. 2017. Development of Cabbage juice medium for industrial production of Leuconostoc mesenteroides starter. J. Microbiol. Biotechnol. 27, 2112-2118. https://doi.org/10.4014/jmb.1708.08050
  7. Jeong, S. J., Yang, H. J., Jeong, S. Y. and Jeong, D. Y. 2015. Identification of characterization and statistical optimization of medium constituent for Bacillus subtilis SCJ4 isolated from Korean traditional fermented food. Kor. J. Microbiol. 51, 48-60. https://doi.org/10.7845/kjm.2015.5004
  8. Joshi, S., Yadav, S., Nerurkar, A. and Desai, A. J. 2007. Statistical optimization of medium components for the production of biosurfactant by Bacillus licheniformis K51. J. Microbiol. Biotechnol. 17, 313-319.
  9. Lee, H. R., Ahn, J. E. and Han, N. S. 2013. Chemical mutation of Leuconostoc mesenteroides for improved mannitol production: development of a high-throughput screening strategy. Biothechnol. Bioprocess Eng. 28, 213-215.
  10. Otgonbayar, G. E., Eom, H. J., Kim, B. S. and Han, N. S. 2011. Mannitol production by Leuconostoc citreum KACC 91348P isolated from Kimchi. J. Microbiol. Biotechnol. 21, 968-971. https://doi.org/10.4014/jmb.1105.05034
  11. Park, E. H., Yoo, B. H. and Kim, M. D. 2018. Characterization of the petent mannitol producer Leuconostoc mesenteroides KCTC13305. J. Agri. Life. Sci. 30, 41-49.
  12. Ryu, B. H., Kim, D. H. and Yun, J. W. 1996. Characteristics of mannitol production by Leuconostoc sp. KY-002. KSBB J. 11, 636-641.
  13. Saitou, N. and Nei, M. 1987. The Neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol. 4, 406-425.
  14. Seo, J. W., Yang, H. J., Jeong, S. J., Ryu, M. S., Ha, G. S., Jeong, S. Y. and Jeong, D. Y. 2018. Characterization of Lactobacillus brevis SCML 432 isolated from Meju in Sunchang and optimization of its culture conditions by statistical methods. Kor. J. Food Preserv. 25, 397-410. https://doi.org/10.11002/kjfp.2018.25.3.397
  15. Yang, H. J., Park, C. S., Yang, H. Y., Jeong, S. J., Jeong, S. Y., Jeong, D. Y., Kang, D, O., Moon, J. Y. and Choi, N. S. 2015. Optimization of medium for the production of cellulase by Bacillus subtilis NC1 using response surface methodology. J. Life Sci. 25, 680-685. https://doi.org/10.5352/JLS.2015.25.6.680
  16. Yoo, S. K., Hur, S. S., Song, S. H., Kimg, K. M. and Whang, K. S. 2005. Optimization of mannitol fermentation by Leuconostoc mesenteroides sp. strain JFY. J. Life Sci. 15, 374-381. https://doi.org/10.5352/JLS.2005.15.3.374
  17. Yue, M., Cao, H., Zhang, J., Meng, Y., Chen, W., Huang, L. and Du, Y. 2013. Improvement of mannitol production by Lactobacillus brevis mutant 3-A5 based on dual-stage pH control and fed-batch fermentations. World J. Microbiol. Biotechnol. 29, 1923-1930. https://doi.org/10.1007/s11274-013-1357-6
  18. Yun, J. W., Kang, S. C., Ryu, B. H. and Song, S. K. 1996. Production of mannitol by Lactobacillus sp. KY-107. KSBB J. 11, 374-379.
  19. Yun, J. W. and Song, S. K. 1994. Mannitol production by Aureobasidium pullurans. KSBB J. 9, 140-146.
  20. Zhang, Z., Cheng, W. Y., Ju, X. Y. and Jin, H. X. 2015. The effect of dextransucrase gene inactivation on mannitol production by Leuconostoc mesenteroides. Indian J. Microbiol. 55, 35-40. https://doi.org/10.1007/s12088-014-0503-7