Detection of Multiple Enzymes in Fermentation Broth Using Single PAGE Analysis
Activity staining or zymography is a technique to detect enzymes based on their function/activity toward a specific substrate. Multiple enzyme-producing microbes secrete enzymes along with other proteins at varying time points during fermentation. The technique of zymography can be used to detect functionality of enzymes in complex protein/other enzyme mixtures. The protein bands corresponding to specific enzyme among other enzymes/proteins can be located by polyacrylamide gel electrophoresis (PAGE) followed by zymogram analysis. This can be employed to locate the secretion pattern of protein/enzyme from intracellular region to extracellular medium. Here we describe simple method for detection and cellular localization of esterases and protease secreted by single microbial strain in one PAGE gel.
Key wordsZymogram Multiple enzymes Fermentation broth PAGE
We thank Department of Biotechnology and Department of Science and Technology (DST), Government of India, for their continuous support to carry out research in our lab. K. Divakar thanks DST, Govt. of India for funding through DST-INSPIRE Faculty award and research grant (IFA14-ENG-87).