Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
205123 | Fuel | 2016 | 7 Pages |
•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.