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Table 2 Amino acid substitutions identified in inactive clones

From: Structural insights from random mutagenesis of Campylobacter jejunioligosaccharyltransferase PglB

Single mutants

Double mutants

Mutation

PglB production

Mutation

PglB production

V432F

strong

  

W457S

strong

W457R-N582S

weak

W458R

strong

  

W458C (3x)

strong

  

D459V (3x)

strong

D459V-A448T

weak

  

D459N-S492C

strong

Y460N

strong

  

Y460C

weak

  

G461D

strong

G461D-S488P

weak

R465H

strong

R465H-G477V

weak

D469L

strong

  

D475F

strong

  

D475Y

strong

  

D475E

weak

  

G476D

weak

G476D-K646I

weak

G481D

strong

  

G481N

strong

  

G481S (2x)

strong

  

S488P (2x)

weak

S488P-V631I

weak

D494V

weak

  

M501K

weak

  

A528V

strong

  

K556N

weak

  

A566S

weak

  

A682D

strong

  
  1. Random mutagenesis of pglB Cj by epPCR was restricted to the region encoding the periplasmic domain (H423-I713). All clones reproducibly exhibited ELISA signals for EPA-CP5 which were ≤ 20% of the wild-type PglB control strain (pGVXN112Kan). Only results for clones which expressed detectable amounts of PglB in DWP are shown (analysed by anti-HA Western blot).
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BMC Biotechnology

ISSN: 1472-6750

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