Isotachophoresis-based sample preparation of cellulases in sugarcane juice using bovine serum albumin as a model protein.
The foremost requirement of quantification of cellulases expressed in genetically modified sugarcane is an efficient sample clean-up. This work investigates the feasibility of isotachophoresis for this purpose. An electrolyte system comprising a leading electrolyte of 10mM formic acid at pH 9.0 and a terminating electrolyte of 10mM β-alanine was devised and used to perform isotachophoresis of cellulases. The use of a simple front cutting method removed a majority of interfering species in the juice, thereby resulting in the formation of a distinct zone of desired proteins. In comparison to techniques such as ultrafiltration and liming, the analysis time and loss of desired proteins was lower when the sample was prepared by using isotachophoresis. Hence, isotachophoresis was an ideal choice for purification of the proteins in question from the remaining components in the juice.
Gupta
Ruchi
R
School of Electrical and Electronics Engineering, The University of Manchester, PO Box 88, Sackville Street Building, Sackville Street, Manchester, M60 1QD, UK. ruchi.gupta@postgrad.manchester.ac.uk
Baldock
Sara J
SJ
Fielden
Peter R
PR
Prest
Jeff E
JE
Grieve
Bruce D
BD
D001628:Beverages / Q000032:analysis; D044602:Cellulases / Q000032:analysis / Q000302:isolation & purification*; D004586:Electrophoresis / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D010940:Plant Proteins / Q000032:analysis / Q000302:isolation & purification*; D012015:Reference Standards; D031786:Saccharum / Q000737:chemistry / Q000201:enzymology*; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000302:isolation & purification*
D010940:Plant Proteins; D012710:Serum Albumin, Bovine; D044602:Cellulases