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Table 1

From: High-throughput screening of cellulase F mutants from multiplexed plasmid sets using an automated plate assay on a functional proteomic robotic workcell

Clone

Plate

Sequence of CelF Terminal Codons

pH Range

50°C, 1 Hour Stability

Mutagenesis

  

NNR

NNR

NNR

NNR

*

  

WT #5 CONTROL

From single 96-clone screen

AAT

GCT

AGA

CCA

GGA

TTC

TAA

5.0–5.8

Moderate

   

Asn

Ala

Arg

Pro

Gly

Phe

*

  

#62 CONTROL

From single 96-clone screen

AAT

GCT

CCA

GTA

CGG

GAA

TAA

5.0–5.8

Moderate

   

Asn

Ala

Pro

Val

Arg

Glu

*

  

COLONY PICKING

 

Variants with High Activity at Low pH

  

MAN

AUTO

          

E7

 

From G2 well of 768-clone multiplex screen

AAT

GCT

TTA

TAA

   

4.0–5.8

Not Stable

   

Asn

Ala

Leu

*

     

F2

G12

From B11 well of 768-clone multiplex screen

AAT

GCT

CAA

TGG

ACA

CCA

TAA

4.0–5.8

Moderate

   

Asn

Ala

Gln

Trp

Thr

Pro

*

  

F9 φ

G4 φ

From G2 well of 768-clone multiplex screen

AAT

GCT

ATG

CCA

ACG

TTA

TAA

4.0–5.8

Very Stable

   

Asn

Ala

Met

Pro

Thr

Leu

*

  

F12

 

From G11 well of 768-clone multiplex screen

AAT

GCT

CCA

TAA

   

4.0–5.8

Moderate

   

Asn

Ala

Pro

*

     

HTS Mutagenesis

  

NNR/Y

NNR/Y

NNR/Y

NNR/Y

*

  
 

C7

From Plate 2 well of 23,424- clone multiplex screen

AAT

GCT

GGG

TTA

GAA

TAG

TAA

4.0–5.8

Very Stable

   

Asn

Ala

Gly

Leu

Glu

*

*

  
  1. φ An additional Thr to Ser mutation at amino acid position 213 in F9 and G4 clones