-amylase-pullulanase enzyme, a dna sequence coding the same,...

Details -amylase-pullulanase enzyme, a dna sequence coding the same,... -amylase-pullulanase enzyme, a dna sequence coding the same,...

C - Chemistry, Metallurgy

12

N

195/1.2, 195/1.2

C12N 15/63 (2006.01) C12N 1/00 (2006.01) C12N 9/28 (2006.01) C12N 9/44 (2006.01) C12N 15/56 (2006.01) C12N 15/70 (2006.01) C12N 15/74 (2006.01) C12P 19/14 (2006.01) C12P 19/16 (2006.01) C12P 19/20 (2006.01)

Patent

CA 2018203

ABSTRACT OF THE DISCLOSURE: A gene from Clostridium thermohydrosulfuricum DSM 3783 encoding a thermos-table .alpha.-amylase-pullulanase enzyme was cloned in Escherichia coli as a 7 kb insert using a lambda-vector. For subcloning pUC18 and pUC19 plasmids were used. Using a C. thermohydrosulfuricum promoter at least ten different size of soluble intracellular thermostable, mostly degenerated polypeptides having both .alpha.-amylase and pullulanase activities and Mr in the range 165 000 to 100 000 were produced. The temperature optimum of the enzyme composed of these degenarated polypepetides was (80-85 °C), some 5 °C lower than the temperature optimum of the extracellular enzyme from the native host, but the heat stability was the same as that of the native enzyme. The enzyme preparation can be used alone or in combination with a glucogenic or a maltogenic enzyme for effective hydrolyses of starch.

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