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TK-PUL, a novel thermo-acidophilic pullulanase from  Thermococcus kodakarensis  and a unique member of glycoside hydrolase family GH13 was successfully produced in  Escherichia coli  grown by fed batch culture in a fermenter and partially purified by simple heat treatment. Specific activity of partially purified TK-PUL was 28 U/mg. Corn starch was successfully liquefied and saccharified using this single enzyme at pH 4.2. Simultaneous liquefaction and saccharification of corn starch by TK-PUL was comparable to Termamyl, a commercially available starch-hydrolyzing industrial enzyme. Both enzymes efficiently hydrolysed corn starch into sugar syrups having major proportions of maltose. TK-PUL performs efficiently at the natural pH of starch (~4.5) in the absence of any metal ions, hence is a potential candidate for starch industry.

amylase

TK-PUL, a pullulan hydrolase type III from  Thermococcus kodakarensis , a potential candidate for simultaneous liquefaction and saccharification of starch