N-acetyllactosamine (LacNAc) is an important lactose-derived molecule which can act as an effective prebiotic. In this study a process for the enzymatic synthesis and downstream purification of LacNAc was designed based on the use of thermostable β-galactosidases from Bacillus circulans (BgaD-D), Thermus thermophilus HB27 or Pyrococcus furiosus (CelB) respectively. Four configurations for the purification stage were simulated; anion-exchange chromatography, an activated charcoal-Celite column, N-acetylglucosamine (GlcNAc) crystallization and an activated charcoal-Celite column, as well as selective crystallization. While the enzyme CelB has greater stability at higher temperatures, this enzyme gives a lower LacNAc yield, leading to significant capital investment. For the design based on the BgaD-D biocatalyst and anion exchange chromatography, recovery of GlcNAc improved the project profitability when the GlcNAc price was greater than $10 per kg. GlcNAc was the main contributor to the raw material costs for most processes, although methanol contributed 72% of these costs for the process based on an activated charcoal column. The use of a crystallizer for GlcNAc separation before this column, reduced this methanol consumption by 73%. The use of selective crystallization proved the best approach, reducing the minimum LacNAc sales price to $2 per gram. The plant was more economic when the acceptor to donor ratio was reduced from 10 to 4 and the lactose concentration increased from 50 mM to 550 mM.
- Economic evaluation
Karimi Alavijeh, M., Meyer, A. S., Gras, S. L., & Kentish, S. E. (2020). Simulation and economic assessment of large-scale enzymatic N-acetyllactosamine manufacture. Biochemical Engineering Journal, 154, . https://doi.org/10.1016/j.bej.2019.107459