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Biophysical and Biochemical Analyses of Cellulase‐Polymer Bioconjugates for Biofuel Applications
Author(s) -
Makaroff Katherine,
Lucius Melissa,
Wright Thai,
Stewart Jamie,
Williams Cameron,
Berberich Jason,
Konkolewicz Dominik,
Page Richard C
Publication year - 2016
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.30.1_supplement.843.1
Subject(s) - cellulase , chemistry , polymer , derivatization , raft , cellulose , polymerization , cellulosic ethanol , lysozyme , combinatorial chemistry , organic chemistry , chromatography , chemical engineering , biochemistry , mass spectrometry , engineering
In recent years, due to high demand, protein‐polymer conjugates have become a more common and important biomaterial. Through specific derivatization strategies, polymers of different lengths and types are conjugated onto proteins. The conjugation of polymers onto proteins has been observed to stabilize proteins, and shift the optimal temperature and pH of proteins. To meet increasing demands for biofuels, the production of cellulosic ethanol will need to be increased greater than 600‐fold by 2022. This has lead to the call for more stable thermophilic cellulases, which catalyze the conversion of cellulose to glucose, a precursor for ethanol. Reversible addition‐fragmentation chain transfer (RAFT) is a common derivatization strategy used to produce protein polymer conjugates with increased stability. We initially optimized the RAFT technique for production of lysozyme‐polymer bioconjugates. A modified version of RAFT that involves grafting‐to and grafting‐from techniques was found to produce active lysozyme‐polymer bioconjugates with increased stability. The modified approach allows for fine control over the type and number of polymers conjugated onto the protein. The derivatization strategy and conditions used in the conjugation of lysozyme, are now being applied to the thermophilic cellulase Fn Cel5a. Results of the RAFT‐polymerization strategy with Fn Cel5a will be discussed with respect to the end goal of producing a cellulase that is more stable under the extreme industrial conditions utilized during biofuel production.