Pullulanase (Pul) is an important amylase that can hydrolyze the α-1,6 glycosidic bonds in polysaccharide macromolecules such as starch and pullulan. To increase the fermentation level of Pul, the Pul from Bacillus deramificans was expressed in Bacillus subtilis WB600. Firstly, the effects of native constitutive promoter, synthetic constitutive promoter, inducible promoters, and auto-induction promoters on Pul expression was investigated. It was showed that the synthetic constitutive promoter P566 achieved 58.1 U/mL of Pul, 96% higher than that of P43. Then, the fermentation medium was optimized using single factor and response surface methodology. The optimal medium was composed of corn steep liquor (14.8 g/L), peptone (7.3 g/L), glucose (17.5 g/L), and Ca2+(1.2 mmol/L). The Pul activity reached 129.4 U/mL when the optimal medium was used. Finally, glucose feeding was used in 5 L fermenter, resulting in 226.4 U/mL of Pul after 53 h of fermentation. These results provide a methodological reference for the production of pullulanase in B.subtilis.
XU Kuidong
,
TONG Yi
,
LI Yi
,
TAO Jin
,
LIANG Yingchao
,
LIU Song
,
LI Jianghua
. Combined strategy to improve the expression of pullulanase in Bacillus subtilis[J]. Food and Fermentation Industries, 2022
, 48(5)
: 8
-14
.
DOI: 10.13995/j.cnki.11-1802/ts.029114
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