A Bacillus megaterium plasmid system for the production, export, and one-step purification of affinity-tagged heterologous levansucrase from growth medium.
A multiple vector system for the production and export of recombinant affinity-tagged proteins in Bacillus megaterium was developed. Up to 1 mg/liter of a His6-tagged or Strep-tagged Lactobacillus reuteri levansucrase was directed into the growth medium, using the B. megaterium esterase LipA signal peptide, and recovered by one-step affinity chromatography.
Malten
Marco
M
Institute of Microbiology, Technical University Braunschweig, Spielmannstrasse 7, D-38106 Braunschweig, Germany.
Biedendieck
Rebekka
R
Gamer
Martin
M
Drews
Ann-Christin
AC
Stammen
Simon
S
Buchholz
Klaus
K
Dijkhuizen
Lubbert
L
Jahn
Dieter
D
D000345:Affinity Labels; D001410:Bacillus megaterium / Q000201:enzymology* / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics; D002846:Chromatography, Affinity; D003470:Culture Media; D015870:Gene Expression; D005798:Genes, Bacterial; D005822:Genetic Vectors; D006602:Hexosyltransferases / Q000096:biosynthesis* / Q000235:genetics* / Q000302:isolation & purification; D052200:Limosilactobacillus reuteri / Q000201:enzymology / Q000235:genetics; D010957:Plasmids / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
D000345:Affinity Labels; D001426:Bacterial Proteins; D003470:Culture Media; C079999:LipA protein, Bacteria; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D006602:Hexosyltransferases; C021513:levansucrase