Continuous production of chitooligosaccharides in a column reactor by the PUF-immobilized whole cell enzymes of Mucor circinelloides IBT-83

Microb Cell Fact. 2024 Sep 28;23(1):258. doi: 10.1186/s12934-024-02529-4.

Abstract

Background: Chitosan oligosaccharides (COS) have great potential for applications in several fields, including agriculture, food industry or medicine. Nevertheless, the large-scale use of COS requires the development of cost-effective technologies for their production. The main objective of our investigation was to develop an effective method of enzymatic degradation of chitosan in a column reactor using Mucor circinelloides IBT-83 cells, immobilized in a polyurethane foam (PUF). These cells serve as a source of chitosanolytic enzymes.

Results: The study revealed that the process of freeze-drying of immobilized mycelium increases the stability of the associated enzymes during chitosan hydrolysis. The use of stabilized preparations as an active reactor bed enables the production of COS at a constant level for 16 reactor cycles (384 h in total), i.e. 216 h longer compared to non-stabilized mycelium. In the hydrolysate, oligomers ranging in structure from dimer to hexamer as well as D-glucosamine were detected. The potential application of the obtained product in agriculture has been verified. The results of phytotests have demonstrated that the introduction of COS into the soil at a concentration of 0.01 or 0.05% w/w resulted in an increase in the growth of Lepidium sativum stem and root, respectively (extensions by 38 and 44% compared to the control sample).

Conclusions: The research has verified that the PUF-immobilized M. circinelloides IBT-83 mycelium, which has been stabilized through freeze-drying, is a promising biocatalyst for the environmentally friendly and efficient generation of COS. This biocatalyst has the potential to be used in fertilizers.

Keywords: Mucor circinelloides; Chitooligosaccharides; Chitosanase; Promoting plant growth; Whole-cell biocatalyst.

MeSH terms

  • Bioreactors*
  • Cells, Immobilized / metabolism
  • Chitosan* / chemistry
  • Chitosan* / metabolism
  • Enzymes, Immobilized / chemistry
  • Enzymes, Immobilized / metabolism
  • Freeze Drying
  • Hydrolysis
  • Mucor* / enzymology
  • Mucor* / metabolism
  • Mycelium / metabolism
  • Oligosaccharides* / biosynthesis
  • Oligosaccharides* / metabolism
  • Polyurethanes / chemistry

Substances

  • Chitosan
  • Oligosaccharides
  • oligochitosan
  • Polyurethanes
  • polyurethane foam
  • Enzymes, Immobilized

Supplementary concepts

  • Mucor circinelloides