Rapid micro-assays for amylolytic activities determination: customization and validation of the tests.

ABSTRACT: High-throughput function-based screening techniques remain the major bottleneck in the novel biocatalysts development pipeline. In the present study, we customized protocols for amylolytic activity determination (Somogyi-Nelson and starch-iodine tests) to micro-volume thermalcycler-based assays (linearity range 60-600 μM of reducing sugar, R² = 0.9855; 0-2 mg/mL of starch, R² = 0.9921, respectively). Exploitation of a thermalcycler enabled rapid and accurate temperature control, further reduction of reagents and samples volumes, and limited evaporation of the reaction mixtures, meeting several crucial requirements of an adequate enzymatic assay. In the optimized micro-volume Somogyi-Nelson protocol, we were able to reduce the time required for high-temperature heating

SUBMITTER: Borkowska M 

PROVIDER: S-EPMC6420461 | biostudies-literature | 2019 Mar

REPOSITORIES: biostudies-literature