Thermophilic recombinant type II pullulanase and application thereof

Abstract

The present disclosure discloses a thermophilic recombinant type II pullulanase and the application thereof, and belongs to the technical field of genetic engineering. The present disclosure obtains a thermophilic recombinant type II pullulanase by heterologously expressing type II pullulanase in Escherichia coli. Its optimum pH is 6.6, it has better pH tolerance under the conditions of pH 5.8-8.0, and its optimum temperature is 95° C. After incubating at 95° C. for 10 h, the remaining enzyme activity is greater than 50%. It can exhibit higher specific enzyme activity under strong reducing conditions. For example, adding DTT to the culture environment can increase the specific enzyme activity of Sumo-PulPy by 237.2%. The present disclosure also provides the combined truncation mutant Δ28N+Δ791C of type II pullulanase Sumo-PulPy. The specific enzyme activity of the enzyme mutant is 32.18±0.92 U/mg, which is 5.99 times as high as that of the wild-type enzyme, thereby having important industrial application value and potential.

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