Rapid determination of the synthetic activity of lipases/esterases via transesterification and esterification zymography

• A new and versatile method to determine synthetic activity by zymography is proposed.
• Specific detection of proteins which perform the synthetic activity desired.
• Zymogram results are confirmed by titration, GC-analysis and NMR.
• This method may be applied for enzyme high-throughput screening.

A new simple and extremely versatile zymography method based on transesterification and esterification reactions was developed in this work. The method consists in building a transesterification or esterification reaction medium according to the goal of the research. Since commercial enzymes and crude extracts commonly consist in pools of proteins, this methodology provides a means to determine which protein are responsible for the enzymatic activity desired. The protocol may be potentially used as a high-throughput screening of lipases or esterases that catalyzes the synthesis reaction for biodiesel and biolubricants production.