TY - JOUR
T1 - Autohydrolysis of plant polysaccharides using transgenic hyperthermophilic enzymes
AU - Montalvo-Rodriguez, Rafael
AU - Haseltine, Cynthia
AU - Huess-LaRossa, Kathy
AU - Clemente, Tom
AU - Soto, Jimmy
AU - Staswick, Paul
AU - Blum, Paul
PY - 2000/10/20
Y1 - 2000/10/20
N2 - Commercial bioprocessing of plant carbohydrates, such as starch or cellulose, necessitates the use of commodity enzyme additives to accelerate polysaccharide hydrolysis. To simplify this procedure, transgenic plant tissues constitutively producing commodity enzymes were examined as a strategy for accelerating carbohydrate bioprocessing. Hyperthermophilic glycosyl hydrolases were selected to circumvent enzyme toxicity, because such enzymes are inactive at plant growth temperatures and are therefore physiologically benign. Transgenic tobacco lines were established that produced either a hyperthermophilic α-glucosidase or a β-glycosidase using genes derived from the archaeon Sulfolobus solfataricus. Western blot and immunoprecipitation analyses were used to demonstrate the presence of recombinant enzymes in plant tissues. Transgenic enzyme levels exhibited an unusual delayed pattern of accumulation while their activities survived plant tissue preservation. Transgenic plant protein extracts released glucose from purified polysaccharide substrates at appreciable rates during incubation in high-temperature reactions. Glucose was also produced following enzymatic treatment of plant extracts enriched for endogenous polysaccharides. Direct conversion of plant tissue into free sugar was evident using whole plant extracts of either transgenic line, and could be significantly accelerated in a synergistic manner by combining transgenic line extracts. (C) 2000 John Wiley and Sons, Inc.
AB - Commercial bioprocessing of plant carbohydrates, such as starch or cellulose, necessitates the use of commodity enzyme additives to accelerate polysaccharide hydrolysis. To simplify this procedure, transgenic plant tissues constitutively producing commodity enzymes were examined as a strategy for accelerating carbohydrate bioprocessing. Hyperthermophilic glycosyl hydrolases were selected to circumvent enzyme toxicity, because such enzymes are inactive at plant growth temperatures and are therefore physiologically benign. Transgenic tobacco lines were established that produced either a hyperthermophilic α-glucosidase or a β-glycosidase using genes derived from the archaeon Sulfolobus solfataricus. Western blot and immunoprecipitation analyses were used to demonstrate the presence of recombinant enzymes in plant tissues. Transgenic enzyme levels exhibited an unusual delayed pattern of accumulation while their activities survived plant tissue preservation. Transgenic plant protein extracts released glucose from purified polysaccharide substrates at appreciable rates during incubation in high-temperature reactions. Glucose was also produced following enzymatic treatment of plant extracts enriched for endogenous polysaccharides. Direct conversion of plant tissue into free sugar was evident using whole plant extracts of either transgenic line, and could be significantly accelerated in a synergistic manner by combining transgenic line extracts. (C) 2000 John Wiley and Sons, Inc.
KW - Bioprocessing
KW - Hydrolases
KW - Hyperthermophiles
KW - Polysaccharides
KW - Tobacco
KW - Transgenes
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U2 - 10.1002/1097-0290(20001020)70:2<151::AID-BIT4>3.0.CO;2-D
DO - 10.1002/1097-0290(20001020)70:2<151::AID-BIT4>3.0.CO;2-D
M3 - Article
C2 - 10972926
AN - SCOPUS:0034693454
SN - 0006-3592
VL - 70
SP - 151
EP - 159
JO - Biotechnology and Bioengineering
JF - Biotechnology and Bioengineering
IS - 2
ER -