David Mangan*, Claudio Cornaggia, Vincent McKie and Barry McCleary
Megazyme International, IDA Business Park, Southern Cross, Road, Bray, Co. Wicklow, Ireland
Corresponding Author: David Mangan, David@megazyme.com, +353-1-2861220
Cellulase (endo-1,4-b-glucanase, EG) is arguably the most important glycosyl hydrolase involved in the saccharification process which is central to cellulosic biofuel production.
Traditionally, EG has been assayed using reducing-sugar procedures with cellulose, soluble cellulose derivatives (e.g. carboxymethyl-cellulose) or mixed-linkage b-glucan as substrates. In the 1980s, soluble, dyed polysaccharides and dyed and cross-linked polysaccharides (insoluble, gelatinous) were introduced and have found widespread use. None of these substrates or assay procedures readily lend themselves to automation. To satisfy this need, a range of end-blocked nitrophenyl-cello-oligosaccharides have been produced and evaluated in the assay of cellulase. A novel EG assay procedure based on an enzyme coupled assay format was launched in 2013[1],[2] and recently, a second generation assay displaying marked improvements has been described.[3] A sensitive, rapid, universally applicable and fully automatable assay is presented. A schematic representation is shown below.
