Hembach, Lea; Cord-Landwehr, Stefan; Landwehr, Stefan Cord-; Moerschbacher, Bruno: Enzymatic production of all fourteen partially acetylated chitosan tetramers using different chitin deacetylases acting in forward or reverse mode
Inhalt
- Enzymes – Origins, heterologous expression and purification.
- Partial deacetylation of the chitin tetramer (A4) using different chitin deacetylases.
- Partial N-acetylation of the chitosan tetramer (D4) using different chitin deacetylases.
- UHPLC-ESI-MS analysis of chitin (COS) and chitosan oligomers (paCOS).
- Determination of the pattern of acetylation of paCOS.
- Medium-scale production and purification of paCOS.
- Table 1 Summarized information about the CDAs used in this study, their origin host as well as the chosen expression host and the protein-tag used for purification (FPLC).
- Results and Discussion
- Figure 1 Relative amounts of different chito-oligosaccharides after performing enzymatic N-acetylation of the chitosan tetramer (D4, [GlcN]4) using different CDAs [BcCDA5, NodB, CvCDA, VcCDA, PesCDA, CnCDA2, PaCDA, CnCDA4, PgtCDA] for 98 h in 2 M sodium a
- Figure 2 Base peak chromatograms of HILIC-ESI-MS analysis showing products after enzymatic deacetylation of the fully acetylated GlcNAc tetramer (A4, [GlcNAc]4, filled circles) (A,C) and after enzymatic N-acetylation of the fully deacetylated GlcN tetrame
- Figure 3 Mode of action of different CDAs (bacterial, viral, and fungal) on the chitin tetramer (A4, [GlcNAc]4, filled circles) shown in the upper part, as well as on the chitosan tetramer (D4, [GlcN]4, open circles) in the lower part.
- Figure 4 Combination of bacterial CDAs during N-acetylation.
- Figure 5 Combination of bacterial and fungal CDAs during N-acetylation.
- Figure 6 Production routes of all possible chitin and chitosan tetramers using CDAs to specifically deacetylate or N-acetylate (pa)COS 1 represents the use of NodB, 2 of VcCDA, 3 of PesCDA, 4 of CnCDA4, and 5 of PgtCDA.
- Conclusion
- Figure 7 Chromatographic purification up to analytical grade of the (pa)COS DDAD.
- Acknowledgements