Novel chemical- and protein-mediated methods for glucosamine detection
Food Materials Research(2022)
摘要
We describe two novel approaches for the determination of glucosamine (GlcN). The first approach is based on the chemical derivatization of GlcN with the non-fluorophor 1,3-diphenyl-1,3-propanedione (DPPD), which results in a condensation product with interesting fluorescent properties. The obtained compound was isolated by silica-gel chromatography and its structure elucidated by NMR and mass spectrometry. The second approach is based on a previously undescribed sensitivity of the enzyme glucosamine-6-phosphate deaminase (GPDA) towards GlcN, which resulted in the precipitation of the enzyme. Using a rational enzyme engineering approach and both liquid-based and plate-based screening methods, mutational GPDA variants with significantly improved precipitation properties were identified and characterized. These novel glucosamine detection methods may be a useful addition to the repertoire of currently available glucosamine detection sensors.
更多查看译文
关键词
glucosamine detection,fluorescence labeling,protein engineering,assay development,directed evolution
AI 理解论文
溯源树
样例
生成溯源树,研究论文发展脉络
Chat Paper
正在生成论文摘要