2002
DOI: 10.1002/bit.10301
|View full text |Cite
|
Sign up to set email alerts
|

Anchorage of cyclodextrin glucanotransferase on the outer membrane of Escherichia coli

Abstract: The gene encoding cyclodextrin glucanotransferase (CGTase) was successfully cloned from B. macerans by PCR. A recombinant plasmid pCS005 with a gene encoding the Lpp-OmpA-CGTase trifusion protein was constructed and transformed into E. coli for the surface display of CGTase. Results of immunoblotting analysis and protease accessibility on the fractionated cell membranes confirmed that the Lpp-OmpA-CGTase trifusion protein was successfully anchored on the outer membrane of E. coli. However, only 50% of the memb… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
1
1

Citation Types

2
23
0

Year Published

2005
2005
2021
2021

Publication Types

Select...
6
1
1

Relationship

0
8

Authors

Journals

citations
Cited by 22 publications
(25 citation statements)
references
References 32 publications
2
23
0
Order By: Relevance
“…The LppOmpA and INP systems used in this study look promising, since they are well suited as a carrier of relatively large inserts. The largest proteins that have been successfully displayed with Lpp-OmpA and INP in E. coli so far are a 74-kDa cyclodextrin glucanotransferase and a 90-kDa chitinase, respectively (42,45).…”
Section: Resultsmentioning
confidence: 99%
See 1 more Smart Citation
“…The LppOmpA and INP systems used in this study look promising, since they are well suited as a carrier of relatively large inserts. The largest proteins that have been successfully displayed with Lpp-OmpA and INP in E. coli so far are a 74-kDa cyclodextrin glucanotransferase and a 90-kDa chitinase, respectively (42,45).…”
Section: Resultsmentioning
confidence: 99%
“…The Lpp-OmpA-based cell display system has been extensively used for the display of heterologous proteins, such as ␤-lactamase (11), cellulases (12), the scFv antibody (6), cyclodextrin glucanotransferase (42), and chitinbinding domain (43), on the surface of E. coli.…”
mentioning
confidence: 99%
“…The mislocalization of target proteins can affect their functions negatively ( Dieye et al, 2003;Van Der Vaart et al, 1997;Wan et al, 2002). In contrast, the surface display systems based on the external mode of protein display can ensure the full exposure of target proteins outside of the cell wall and the surface intensity of target proteins can readily be adjusted by selecting appropriate display hosts and suitable concentrations for the fusion proteins in the incubation mixture.…”
Section: Anchoring Domains In Surface Display Systems Of Labmentioning
confidence: 99%
“…Lpp-OmpA is an efficient surface display system developed by Georgiou et al (9), which has been used successfully to anchor a variety of proteins, including some enzymes, onto the cell surface (10,21). This display system allows C-terminal fusion of the passenger proteins and consists of two key anchoring motifs: (i) the signal sequence and the first nine amino acids of E. coli lipoprotein (Lpp), to target the proteins to the inner face of the outer membrane, and (ii) the transmembrane region (amino acids 46 to 159) of E. coli outer membrane protein A (OmpA), to conduct the proteins across the outer membrane.…”
mentioning
confidence: 99%
“…This display system allows C-terminal fusion of the passenger proteins and consists of two key anchoring motifs: (i) the signal sequence and the first nine amino acids of E. coli lipoprotein (Lpp), to target the proteins to the inner face of the outer membrane, and (ii) the transmembrane region (amino acids 46 to 159) of E. coli outer membrane protein A (OmpA), to conduct the proteins across the outer membrane. Since the Lpp-OmpA mode combines the anchoring capabilities of lipoprotein and outer membrane protein, it has outstanding advantages in high surface display efficiency and strong adaptability to passenger proteins varying in size (21).…”
mentioning
confidence: 99%