Rapid determination of the synthetic activity of lipases/esterases via transesterification and esterification zymography. (1st August 2016)

• Record Type: Journal Article
• Authors: Duarte, Jaqueline G.; Leone-Ignacio, Kassia; da Silva, Jose Andre C.; Fernandez-Lafuente, Roberto; Freire, Denise M.G.
• Is Part Of: Fuel. Volume 177(2016)
• Page Start: 123
• Page End: 129
• Publication Date: 2016-08-01
• Subjects: Transesterification reactions -- Zymography -- Biodiesel -- Lipase -- Biolubricants; Fuel -- Periodicals; Coal -- Periodicals; Coal; Fuel; Periodicals; 662.6
• DOI: 10.1016/j.fuel.2016.02.079
• Languages: English
• ISSNs: 0016-2361

Abstract

Highlights: 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. Abstract: 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.