2015
DOI: 10.1128/aem.03514-14
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A Vector System for ABC Transporter-Mediated Secretion and Purification of Recombinant Proteins in Pseudomonas Species

Abstract: Pseudomonas fluorescens is an efficient platform for recombinant protein production. P. fluorescens has an ABC transporter secreting endogenous thermostable lipase (TliA) and protease, which can be exploited to transport recombinant proteins across the cell membrane. In this study, the expression vector pDART was constructed by inserting tliDEF, genes encoding the ABC transporter, along with the construct of the lipase ABC transporter recognition domain (LARD), into pDSK519, a widely used shuttle vector. When … Show more

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Cited by 16 publications
(25 citation statements)
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“…2A and supplemental Table S1). Similarly intriguing, other reports of recombinant protein secretion through ABC transporters often, if not always, involve acidic, negatively-charged target proteins: mannanase (13), phospholipase A 1 , 3 alkaline phosphatase (11), and epidermal growth factor (10). We also added the pI values of these proteins in Fig.…”
Section: Cross-analyzing the Secretion Of Recombinant Proteins And Thmentioning
confidence: 99%
See 2 more Smart Citations
“…2A and supplemental Table S1). Similarly intriguing, other reports of recombinant protein secretion through ABC transporters often, if not always, involve acidic, negatively-charged target proteins: mannanase (13), phospholipase A 1 , 3 alkaline phosphatase (11), and epidermal growth factor (10). We also added the pI values of these proteins in Fig.…”
Section: Cross-analyzing the Secretion Of Recombinant Proteins And Thmentioning
confidence: 99%
“…Plasmid pDART was used for the secretory production of different proteins (11). Plasmid vectors pFD10 and pBD10 were derivatives of pDART, constructed by adding codons for 10 aspartic acid residues to the target proteins in either the upstream or downstream position of MCS.…”
Section: Plasmid Vector Constructionsmentioning
confidence: 99%
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“…Simple addition of AP‐secreting microbial cells generated the phosphatase activity incorporated BIOMOSAIC films. The observed dephosphorylation from para ‐nitrophenyl phosphate was to produce 160 × 10 −6 m of para ‐nitrophenol in 24 h (Figure f) . The reusability of BIOMOSAIC–AP film tested by 3 consecutive times of the film incubation was that about 50.6 ± 0.5% activity was remained (Figure S13, Supporting Information).…”
Section: Resultsmentioning
confidence: 95%
“…Phosphate Hydrolysis Using BIOMOSAIC–AP Film : The biocatalytic activity of the BIOMOSAIC–AP film was determined using para ‐nitrophenyl phosphate ( p ‐NPP) as a substrate . The BIOMOSAIC–AP film (3.5 cm × 3.5 cm) was incubated with 25 mL of 5 × 10 −3 m p ‐NPP in a 0.1 m glycine buffer 100 × 10 −3 m Tris‐HCl (pH 8), 100 × 10 −3 m NaCl, and a 5 × 10 −3 m MgCl 2 buffer.…”
Section: Methodsmentioning
confidence: 99%