화학공학소재연구정보센터
Journal of Industrial and Engineering Chemistry, Vol.115, 528-536, November, 2022
Aqueous two-phase extraction to separate alpha-ketoglutaric acid from fermentation broth by combining crystallization
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Alpha-ketoglutaric acid (a-KG) is a crucial intermediate with broad application prospects. However, the downstream separation of a-KG from fermentation broth with low concentration and complex components has become the bottleneck of large-scale production. In this study, a-KG was separated from the actual fermentation broth by aqueous two-phase system (ATPS) and crystallization. The optimal ATPS composed of acetone/(NH4)2SO4 was selected by studying the phase diagram and a-KG distribution behavior in different organic solvents and salts/sugars. Under the optimized conditions, the purity of a-KG products from crude fermentation broth reached 98.08%, and the one-pass extraction efficiency was 90.52%. The removal of cell debris and soluble proteins increased with the increase of acetone and (NH4)2SO4 concentrations, with the removal ratios reaching up to 99.44% and 86.54%. In addition, 92.56% of the glucose was distributed in the aqueous phase. Although the a-KG fermentation broth is more complex than the whole-cell biotransformation, impurities in the crude fermentation broth could be efficiently removed and a higher a-KG distribution coefficient was obtained. Meanwhile, the recovered (NH4)2SO4 could be reused for fermentation medium. This strategy provides an economical and effective method combined with upstream biosynthesis to recover a-KG and other bioproducts from crude fermentation broth.
  1. Zeng WZ, Zhang HL, Xu S, Fang F, Zhou JW, Bioresour. Technol., 243, 1037 (2017)
  2. Tischler J, Gruhn WJ, Reid J, Allgeyer E, Buettner F, Marr C, et al., Embo J., 38, e99518 (2018)
  3. Gyanwali B, Lim ZX, Soh J, Lim C, Guan SP, Goh J, et al., Trends Endocrinol. Met., 33, 136 (2022)
  4. Luo ZS, Yu SQ, Zeng WZ, Zhou JW, Biotechnol. Adv., 47, e107706 (2021)
  5. Bayliak MM, Lushchak VI, Ageing Res. Rev., 66, e101237 (2021)
  6. Zdzisinska B, Zurek A, Kandefer-Szerszen M, Arch. Immunol. Ther. Ex., 65, 21 (2017)
  7. Beer B, Pick A, Sieber V, Metab. Eng., 40, 5 (2017)
  8. Xie SQ, Li ZX, Zhu GD, Song WL, Yi CH, J. Clean Prod., 343, e131033 (2022)
  9. Zhang XD, Xu N, Li JL, Ma ZP, Wei L, Liu QD, et al., Enzyme Microb. Technol., 136, e109530 (2020)
  10. Cybulski K, Tomaszewska-Hetman L, Rakicka M, Laba W, Rymowicz W, Rywinska A, Ind. Crop. Prod., 119, 102 (2018)
  11. Song Y, Li JH, Shin HD, Liu L, Du GC, Chen J, Bioresour. Technol., 219, 716 (2016)
  12. Wieschalka S, Blombach B, Bott M, Eikmanns BJ, Appl. Microbiol. Biotechnol., 94, 449 (2012)
  13. Trinh LTP, Lee YJ, Park CS, Bae HJ, J. Ind. Eng. Chem., 69, 57 (2019)
  14. Morgunov IG, Kamzolova SV, Samoilenko VA, Appl. Microbiol. Biotechnol., 97, 8711 (2013)
  15. Szczygielda M, Prochaska K, J. Membr. Sci., 536, 37 (2017)
  16. Peng XY, Zeng WZ, Xu S, Zha J, Zhou JW, Xu GQ, Biotechnol. Prog., 34, 1370 (2018)
  17. Pal D, Thakre N, Kumar A, Keshav A, Sep. Sci. Technol., 51, 1141 (2016)
  18. Inci I, Asçı YS, Uslu H, J. Ind. Eng. Chem., 18, 152 (2012)
  19. Razmara RS, Daneshfar A, Sahrai R, J. Ind. Eng. Chem., 17, 533 (2011)
  20. Xie SQ, Song WL, Yi CH, Qiu XQ, J. Ind. Eng. Chem., 56, 145 (2017)
  21. Zhang ZR, Shen JT, Dai JY, Sun YQ, Dong YS, Xiu ZL, Sep. Purif. Technol., 242, e116784 (2020)
  22. Fu HX, Wang XD, Sun YQ, Yan L, Shen JT, Wang JF, et al., Sep. Purif. Technol., 180, 44 (2017)
  23. Baral P, Pundir A, Kurmi A, Singh R, Kumar V, Agrawal D, Sep. Purif. Technol., 269, e118788 (2021)
  24. Sun YQ, Yan L, Fu HX, Xiu ZL, Process Biochem., 49, 506 (2014)
  25. Li H, Liu HX, Fang J, Yin S, Chen CY, Li CL, J. Chem. Technol. Biotechnol., 94, 3850 (2019)
  26. Beltran AB, Nisola GM, Vivas EL, Cho W, Chung WJ, J. Ind. Eng. Chem., 19, 182 (2013)
  27. Qi LB, Hu YX, Chai QZ, Wang Q, J. Ind. Eng. Chem., 72, 346 (2019)
  28. Shi XQ, Yang M, Chu AQ, Zhang FY, Fang J, Xu N, et al., AIChE J., e17814 (2022)
  29. Shi XQ, Liu HX, Chu AQ, Yang M, Fang J, Yi S, et al., Process Biochem., 111, 241 (2021)
  30. Dai JY, Liu CJ, Xiu ZL, Process Biochem., 50, 1951 (2015)
  31. Dai JY, Ma LH, Wang ZF, Guan WT, Xiu ZL, Bioprocess. Biosyst. Eng., 40, 423 (2017)
  32. Liu JX, Guo YB, Xu XF, J. Pharm. Res., 30, 335 (2011)
  33. Bradford MM, Anal. Biochem., 72, 248 (1976)
  34. Chen X, Zhang DJ, Qi WT, Gao SJ, Xiu ZL, Xu P, Appl. Microbiol. Biotechnol., 63, 143 (2003)
  35. Sun YQ, Zhang XX, Zheng YF, Yan L, Xiu ZL, Sep. Purif. Technol., 209, 972 (2019)
  36. Fu HX, Sun YQ, Teng H, Zhang DJ, Xiu ZL, Sep. Purif. Technol., 139, 36 (2015)