Efficient Secretory Expression of Hyperthermoacidophilic α-Amylase PFA in Bacillus subtilis WB600

doi:10.7506/spkx1002-6630-201818016

Abstract

Abstract: In this study, we screened 9 secretory (Sec) pathway signal peptides for the production of recombinant hyperthermoacidophilic α-amylase PFA from Pyrococcus furiosus in Bacillus subtilis WB600. The signal peptide YfkN achieved the highest extracellular α-amylase activity. Then we used saturation mutagenesis of Ile3 and Gln4 of the signal peptide YfkN as a novel approach to improve the secretion of PFA. The culture supernatant of the recombinant strain contained the signal peptide I3G/Q4R and had remarkable α-amylase activity (up to 715 U/mL). The purified α-amylase PFA showed maximal activity at pH 5.0 and 100 ℃, and its half-life at 100 ℃ was about 13 h with or without 5 mmol/L Ca2+ added. To the best of our knowledge, we obtained higher yield of PFA in a food-grade host, which may benefit the industrial production and application of PFA.

Key words: hyperthermoacidophilic α-amylase, Bacillus subtilis, signal peptide, secretory expression, saturation mutagenesis

CLC Number:

Q814

YUAN Lin, ZENG Jing, GUO Jianjun, GUO Hao, YANG Gang, CHEN Jun. Efficient Secretory Expression of Hyperthermoacidophilic α-Amylase PFA in Bacillus subtilis WB600[J]. FOOD SCIENCE, 2018, 39(18): 100-108.

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Efficient Secretory Expression of Hyperthermoacidophilic α-Amylase PFA in Bacillus subtilis WB600

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