Development, characterization and application of 3D printed adsorbents for in situ recovery of taxadiene from microbial cultivations

J Chromatogr A. 2024 Apr 26:1721:464815. doi: 10.1016/j.chroma.2024.464815. Epub 2024 Mar 19.

Abstract

Microbial cell factories are an attractive alternative to produce high-value natural products using sustainable processes. However, product recovery is one of the main challenges to reduce production cost and make these technologies economically interesting. In this work, new resins were formulated to 3D print hydrophobic adsorbents for the recovery of biologics from microbial cultivations. Benzyl methacrylate (BEMA) and butyl methacrylate (BUMA) were selected as functional monomers suitable for the adsorption of hydrophobic compounds. Pore morphology was tailored through the inclusion of pore forming agents (porogens) in the resin. Different porogens and porogen concentrations were evaluated resulting in materials with different porous networks. Sudan 1 and the anticancer drug paclitaxel were employed as model compounds to test the adsorption performance of hydrophobic and terpene molecules onto the developed 3D printed materials. The material with greatest adsorption capacity was obtained using BEMA monomer with 40 % (v/v) porogen (BEMA40). The performance of BEMA40 to recover taxadiene from small-scale (5 mL) Saccharomyces cerevisiae cultivations was tested and compared with commercial Diaion HP-20 beads. Taxadiene titres on BEMA40 (46 ± 2 mg/L) and Diaion HP-20 (54 ± 4 mg/L) were comparable, with no taxadiene detected in the cells and cell-free media, suggesting near 100 % taxadiene partition on the adsorbents. Compared to commercial beads, 3D printed adsorbents can be customized with adjustments in the resin formulation, are well adaptable to diverse bioreactor types, do not clog sampling ports and columns and are easier to handle during post processing. The results of this work demonstrate the potential of 3D printing to fabricate hydrophobic interaction adsorbent materials and their application in the recovery of biological products.

Keywords: 3D printing; Downstream processing; In situ product recovery; Saccharomyces cerevisiae; Solid adsorption; Taxadiene.

MeSH terms

  • Alkenes*
  • Diterpenes* / chemistry
  • Methacrylates*
  • Paclitaxel
  • Printing, Three-Dimensional
  • Saccharomyces cerevisiae / metabolism
  • Terpenes

Substances

  • taxa-4(5),11(12)diene
  • Diterpenes
  • Paclitaxel
  • Terpenes
  • benzyl methacrylate
  • Alkenes
  • Methacrylates