Doubling Power Output of Starch Biobattery Treated by the Most Thermostable Isoamylase from an Archaeon Sulfolobus tokodaii.
Biobattery, a kind of enzymatic fuel cells, can convert organic compounds (e.g., glucose, starch) to electricity in a closed system without moving parts. Inspired by natural starch metabolism catalyzed by starch phosphorylase, isoamylase is essential to debranch alpha-1,6-glycosidic bonds of starch, yielding linear amylodextrin - the best fuel for sugar-powered biobattery. However, there is no thermostable isoamylase stable enough for simultaneous starch gelatinization and enzymatic hydrolysis, different from the case of thermostable alpha-amylase. A putative isoamylase gene was mined from megagenomic database. The open reading frame ST0928 from a hyperthermophilic archaeron Sulfolobus tokodaii was cloned and expressed in E. coli. The recombinant protein was easily purified by heat precipitation at 80 (o)C for 30 min. This enzyme was characterized and required Mg(2+) as an activator. This enzyme was the most stable isoamylase reported with a half lifetime of 200 min at 90 (o)C in the presence of 0.5 mM MgCl2, suitable for simultaneous starch gelatinization and isoamylase hydrolysis. The cuvett-based air-breathing biobattery powered by isoamylase-treated starch exhibited nearly doubled power outputs than that powered by the same concentration starch solution, suggesting more glucose 1-phosphate generated.
College of Life Sciences, Henan Agricultural University, 95 Wenhua Road, Zhengzhou, 450002, China.|Biological Systems Engineering Department, Virginia Tech, 304 Seitz Hall, Blacksburg, Virginia 24061, USA.
Kun
K
Cheng
Cell Free Bioinnovations Inc. 1800 Kraft Drive, Suite 222, Blacksburg, Virginia 24060, USA.
Fei
F
Zhang
Cell Free Bioinnovations Inc. 1800 Kraft Drive, Suite 222, Blacksburg, Virginia 24060, USA.
Fangfang
F
Sun
College of Life Sciences, Henan Agricultural University, 95 Wenhua Road, Zhengzhou, 450002, China.
Hongge
H
Chen
Biological Systems Engineering Department, Virginia Tech, 304 Seitz Hall, Blacksburg, Virginia 24061, USA.|Cell Free Bioinnovations Inc. 1800 Kraft Drive, Suite 222, Blacksburg, Virginia 24060, USA.|Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, China.
Y-H
YH
Percival Zhang
D000595:Amino Acid Sequence; D000687:Amylopectin / Q000737:chemistry; D055162:Biocatalysis; D001674:Bioelectric Energy Sources; D020134:Catalytic Domain; D017124:Conserved Sequence; D003912:Dextrins / Q000737:chemistry; D004560:Electricity; D004591:Electrophoresis, Polyacrylamide Gel; D004795:Enzyme Stability; D006868:Hydrolysis; D007455:Iodine / Q000737:chemistry; D007517:Isoamylase / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D013213:Starch / Q000737:chemistry*; D016944:Sulfolobus / Q000201:enzymology*; D013696:Temperature
D003912:Dextrins; D013213:Starch; C010026:amylodextrins; D000687:Amylopectin; D007455:Iodine; D007517:Isoamylase