Quantitative determination of cellulase concentration as distinct from cell concentration in studies of microbial cellulose utilization: analytical framework and methodological approach.
In analyzing microbial cellulose utilization, it would be useful to independently measure the mass concentration of cells and cellulase enzymes. Such measurements would allow investigation of the allocation of cellular resources between synthesis of cells and cellulase, in vivo cell- and cellulase-specific cellulose hydrolysis rates, and bioenergetics. Methodological protocols are not established for independent determination of cell and cellulase concentrations for the common case in which a substantial fraction of cellulase is attached to the cell surface. Alternative analytical approaches by which to develop such protocols are examined from the perspective of error minimization. For cell concentration measurement, acceptable accuracy is expected when the concentrations of a cell-specific component (e.g., DNA) is determined or when total protein is determined in conjunction with a measurement specific to cellulase. For cellulase concentration measurement, acceptable accuracy is expected when a measurement specific to cellulase such as ELISA is used. Several analytical approaches are rejected based on large expected errors.
Lynd
Lee R
LR
Chemical and Biochemical Engineering, Thayer School of Engineering, Dartmouth College, Hanover, New Hampshire 03755, USA. Lee.Lynd@dartmouth.edu
Zhang
Yiheng
Y
D001419:Bacteria / Q000201:enzymology / Q000254:growth & development*; D015982:Bias; D002480:Cellulase / Q000032:analysis* / Q000378:metabolism; D002482:Cellulose / Q000378:metabolism*; D004734:Energy Metabolism; D005069:Evaluation Studies as Topic; D005658:Fungi / Q000201:enzymology / Q000254:growth & development*
D002482:Cellulose; D002480:Cellulase