Abstract
Objective
To explore an efficient use of crude glycerol for the production of a highly thermostable β-mannanase (ReTMan26) by Pichia pastoris X33.
Results
Cell growth was significantly inhibited by 4 and 6% (w/v) crude glycerol in 250 ml shake-flasks and in 5 l bioreactor batch cultures, respectively, but not affected by pure glycerol at the same concentrations. For further study, the impact of various impurities in crude glycerol on the cell growth of, and ReTMan26 production by, Pichia pastoris was investigated. Salts and methanol did not exert an inhibitory effect, but ≥ 0.2% and 0.3% (w/v) soap in shake-flask and bioreactor cultures, respectively, inhibited fermentation. Under identical conditions, the biomass and ReTMan26 activity produced by high-cell-density fermentation using 5% crude glycerol (glycerol at 80%, w/w) were slightly higher than those using 4% (w/v) pure glycerol.
Conclusions
Non-pretreated ≤ 5% (w/v) crude glycerol could be effectively utilized for industrial production of ReTMan26, and the total production costs using crude glycerol were ~ 4.2% lower than those using pure glycerol.
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Acknowledgements
This work was supported by the open project of Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University (Grant No.KLIB-KF201510), and the Fundamental Research Funds for the Central Universities (Grant No. JUSRP51504 & 111A24), the 111 Project (No. 111-2-06).
Supporting information
Supplementary Table 1—Composition of crude glycerol used in this work.
Supplementary Table 2—The contents of dioxins and mycotoxins in crude glycerol.
Supplementary Table 3—Effect of salt with various concentrations on cell growth and ReTMan26 production.
Supplementary Figure 1—Metabolic pathways of glycerol utilized by microorganisms.
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Luo, Z., Miao, J., Luo, W. et al. Crude glycerol from biodiesel as a carbon source for production of a recombinant highly thermostable β-mannanase by Pichia pastoris . Biotechnol Lett 40, 135–141 (2018). https://doi.org/10.1007/s10529-017-2451-x
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DOI: https://doi.org/10.1007/s10529-017-2451-x