Presentation Information
[P03-303]High-Throughput Mass Spectrometry Methods for Enzyme Activity Screening in Food and Biotechnology Applications
○Wei Zhe Teo1, Ping Han1, Elvis Teng Chua1, Wen Shan Yew1, Chueh Loo Poh1 (1. National University of Singapore (Singapore))
Keywords:
High Throughput Screening,Biofoundry,Automation
Enzyme engineering is a critical research area with applications spanning sustainable food production, nutritional enhancement, and industrial innovation. Huge enzyme libraries are usually generated, and screening these samples require rapid and efficient analytical methods. High-throughput mass spectrometry platforms offer powerful solutions for characterizing diverse enzyme-substrate reactions across a wide range of analytes, from polar metabolites to non-polar lipids.
Two complementary semi-automated workflows involving liquid handlers were developed to screen enzyme activities targeting different subrate classes. Agilent RapidFire (RF) coupled to mass spectrometer was used for analyzing lipids while LC-MS was established to profile enzymatic hydrolysis of complex carbohydrate substrates.
The RF-MS platform successfully analyzed 96-well plates within 30 minutes, demonstrating versatility across analytes of varying polarity. Enzyme activity comparisons revealed that Enzyme B produced higher levels of free fatty acid than Enzyme A, with water presence enhancing hydrolysis efficiency. For the complex carbohydrate substrates, the LC-MS workflow enabled rapid screening of enzyme cocktails, providing detailed compositional insights into hydrolysate profiles and identifying effective combinations for improving fiber solubilization.
These high-throughput MS-based screening platforms enable efficient evaluation of enzyme libraries and reaction conditions for diverse food and biotechnology applications. The workflows are adaptable to various analyte classes and provide actionable data for optimizing enzyme performance in sustainable food processing.
Two complementary semi-automated workflows involving liquid handlers were developed to screen enzyme activities targeting different subrate classes. Agilent RapidFire (RF) coupled to mass spectrometer was used for analyzing lipids while LC-MS was established to profile enzymatic hydrolysis of complex carbohydrate substrates.
The RF-MS platform successfully analyzed 96-well plates within 30 minutes, demonstrating versatility across analytes of varying polarity. Enzyme activity comparisons revealed that Enzyme B produced higher levels of free fatty acid than Enzyme A, with water presence enhancing hydrolysis efficiency. For the complex carbohydrate substrates, the LC-MS workflow enabled rapid screening of enzyme cocktails, providing detailed compositional insights into hydrolysate profiles and identifying effective combinations for improving fiber solubilization.
These high-throughput MS-based screening platforms enable efficient evaluation of enzyme libraries and reaction conditions for diverse food and biotechnology applications. The workflows are adaptable to various analyte classes and provide actionable data for optimizing enzyme performance in sustainable food processing.
Comment
To browse or post comments, you must log in.Log in