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OwnerNasir Ahmed, Naeem Rashid +2inventors
InventorsNasir Ahmed, Naeem Rashid, Muhammad Saleem Haider, Muhammad Akhtar
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A novel thermoacidophilic pullulanase (Tk-PUL) from hyperthermophilic archaeon Thermococcus kodakaraensis KOD1 is described here that efficiently hydrolyzes starch under industrial conditions in the absence of any additional metal ions. The gene encoding Tk-PUL was cloned and expressed in E. coli cells. The purified recombinant enzyme possesses the following properties; shows both pullulanase and α-amylase activities displays highest activity at 95-100° C. active over a broad pH range (3.0-8.5) with optimum working pH 3.5 stable for several hours at 90° C. and displays a half-life of 45 minutes at 100° C. activity and stability are independent of calcium and other metal ions hydrolyzes maltotriose Moreover, recombinant Tk-PUL can be used for single step liquefaction and saccharification of corn starch (without any α-amylase or β-amylase) at pH 4.2 in the absence of calcium.
1-17. (canceled)
A process comprising simultaneous liquefaction and saccharification of starch comprising:
A process comprising liquefaction of a starch solution in the presence of a pullulan hydrolase comprising an amino acid sequence having at least 65%, 85%, 90%, 98% or 99% homology to the amino acid sequence of SEQ ID NO:2 at a pH of about 4.2 in the absence of calcium followed by saccharification of the liquefied starch by Aspergillus niger glucoamylase without pH adjustment.
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