title: Influence of promoter and signal peptide on the expression of pullulanase in Bacillus subtilis.
abstract: To achieve efficient expression and secretion of a biologically-active pullulanase, the effect of promoter and signal peptide on the production of pullulanase was studied. Three types of promoters (PP43, P apr and P amy ) and four types of signal peptides (SP sacB , SP amy , SP aprl and SP aprs ) were combined to construct twelve expression cassettes for pullulanase in Bacillus subtilis. The pullulanase activity assay was employed to quantify the level of differential expression, and a real-time PCR assay was applied to comparatively track the transcriptional level. Under the same experimental conditions, the potency ratios among the three promoters were P apr  > P amy  > PP43. The secretion efficiency ratios mediated by the signal peptides were SP sacB  > SP amy  > SP aprs  > SP aprl . The highest yield of pullulanase could be achieved under the promotion mediated by P apr and secretion by SP sacB .
authors.affiliation: Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin, 300457, People's Republic of China, wypnes@163.com.
authors.forename: Yapin
authors.initials: Y
authors.lastname: Wang
authors.forename: Yihan
authors.initials: Y
authors.lastname: Liu
authors.forename: Zhengxiang
authors.initials: Z
authors.lastname: Wang
authors.forename: Fuping
authors.initials: F
authors.lastname: Lu
mesh_terms: D001412:Bacillus subtilis / Q000235:genetics* / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D006026:Glycoside Hydrolases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals; D021381:Protein Transport; D060888:Real-Time Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D006026:Glycoside Hydrolases; C100181:pullulanase