研究目的
To assess if laccases from different origins had the same or differentiating reaction patterns and hence FTIR fingerprints on a set of naturally occurring phenols including lignin mono-phenols.
研究成果
FTIR represents a method with high potential for establishing a rapid and versatile enzyme assessment method when comparing large populations of laccases for instance during protein engineering.
研究不足
The sensitivity limitation observed in FTIR may be overcome by adjusting other process parameters like reaction time and enzyme loading.
1：Experimental Design and Method Selection:
FTIR was used to monitor enzymatic oxidation of monolignols, sinapic acid, and sinapic aldehyde by four different laccases. PARAFAC and PCA were used for data modelling and quantification.
2：Sample Selection and Data Sources:
Four laccases from different microorganisms were studied, and their activity was assessed by monitoring the oxidation of syringaldazine.
3：List of Experimental Equipment and Materials:
FTIR instrument MilkoScan? FT2, deuterium oxide, sodium acetate buffer, syringaldazine, and the four laccases.
4：Experimental Procedures and Operational Workflow:
Each reaction mixture was injected directly into the cuvette and 50 spectra were acquired consecutively for a total of 15 minutes.
5：Data Analysis Methods:
PARAFAC and PCA were used to analyze the spectral data and establish enzyme activity calibrations.
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