Enzymatic cascades called one-pot multienzyme (OPME) reaction systems enable efficient synthesis of otherwise-challenging carbohydrate targets, and IMCS is commercializing several OPME systems to facilitate its adoption by non-specialists. Storage stability is an important part of any enzyme commercialization effort, but it is particularly important for OPME kits which require carefully determined ratios of enzymes. Here we present our progress in developing stable storage conditions for several recombinant enzymes using intrinsic fluorescence to screen protein stability, accelerated aging studies, and six-month stability studies.  Liquid formulations were developed for seven bacterial enzymes, including an inorganic pyrophosphatase, CMP-sialic acid synthetase, sialic acid aldolase, and four sialyltransferases. These enzymes displayed melting temperature and aggregation temperature improvements of 10-20°C after three rounds of formulation screening. Accelerated aging studies and K<sub>D</sub>/B<sub>22</sub> measurements were used to make final determinations between lead formulations. Two enzymes have successfully completed six-month stability studies at -20°C, 4°C, and room temperature. These results demonstrate the value of liquid formulation optimization and the stability of IMCS’s glycoenzyme products.

Liquid formulation optimization for stability and activity retention of bacterial enzymes

Where thousands of chemistry professionals meet to share ideas and advance scientific and technical knowledge. Spring 2023 abstracts, posters, and presentations will launch at the end of March.

ACS Spring 2023

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.