PMID- 18799474
OWN - NLM
STAT- MEDLINE
DCOM- 20081126
LR  - 20081017
IS  - 1741-0134 (Electronic)
IS  - 1741-0126 (Linking)
VI  - 21
IP  - 11
DP  - 2008 Nov
TI  - Thermostable variants constructed via the structure-guided consensus method also 
      show increased stability in salts solutions and homogeneous aqueous-organic 
      media.
PG  - 673-80
LID - 10.1093/protein/gzn048 [doi]
AB  - Enzyme instability is a major factor preventing widespread adoption of enzymes 
      for catalysis. Stability at high temperatures and in the presence of high salt 
      concentrations and organic solvents would allow enzymes to be employed for 
      transformations of compounds not readily soluble in low temperature or in purely 
      aqueous systems. Furthermore, many redox enzymes require costly cofactors for 
      function and consequently a robust cofactor regeneration system. In this work, we 
      demonstrate how thermostable variants developed via an amino acid sequence-based 
      consensus method also showed improved stability in solutions with high 
      concentrations of kosmotropic and chaotropic salts and water-miscible organic 
      solvents. This is invaluable to protein engineers since deactivation in salt 
      solutions and organic solvents is not well understood, rendering a priori design 
      of enzyme stability in these media difficult. Variants of glucose 1-dehydrogenase 
      (GDH) were studied in solutions of different salts along the Hofmeister series 
      and in the presence of varying amounts of miscible organic solvent. Only the most 
      stable variants showed little deactivation dependence on salt-type and salt 
      concentration. Kinetic stability, expressed by the deactivation rate constant 
      k(d,obs), did not always correlate with thermodynamic stability of variants, as 
      measured by melting temperature T(m). However, a strong correlation (R(2) > 0.95) 
      between temperature stability and organic solvent stability was found when 
      plotting T(50)(60) versus C(50)(60) values. All GDH variants retained stability 
      in homogeneous aqueous-organic solvents with >80% v/v of organic solvent.
FAU - Vazquez-Figueroa, E
AU  - Vazquez-Figueroa E
AD  - School of Chemical and Biomolecular Engineering, Parker H. Petit Institute for 
      Bioengineering and Bioscience, Atlanta, GA 30332-0363, USA.
FAU - Yeh, V
AU  - Yeh V
FAU - Broering, J M
AU  - Broering JM
FAU - Chaparro-Riggers, J F
AU  - Chaparro-Riggers JF
FAU - Bommarius, A S
AU  - Bommarius AS
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20080916
PL  - England
TA  - Protein Eng Des Sel
JT  - Protein engineering, design & selection : PEDS
JID - 101186484
RN  - 0 (Organic Chemicals)
RN  - 0 (Salts)
RN  - 0 (Solutions)
RN  - 0 (Solvents)
RN  - EC 1.1.1.47 (Glucose 1-Dehydrogenase)
SB  - IM
MH  - Animals
MH  - Bacillus subtilis
MH  - Consensus Sequence
MH  - Enzyme Stability/drug effects
MH  - Glucose 1-Dehydrogenase/*chemistry/*genetics
MH  - Hot Temperature
MH  - Kinetics
MH  - Organic Chemicals/chemistry
MH  - Protein Denaturation
MH  - Salts/chemistry/*pharmacology
MH  - Solutions/chemistry
MH  - Solvents/chemistry/*pharmacology
MH  - Temperature
EDAT- 2008/09/19 09:00
MHDA- 2008/12/17 09:00
CRDT- 2008/09/19 09:00
PHST- 2008/09/19 09:00 [pubmed]
PHST- 2008/12/17 09:00 [medline]
PHST- 2008/09/19 09:00 [entrez]
AID - gzn048 [pii]
AID - 10.1093/protein/gzn048 [doi]
PST - ppublish
SO  - Protein Eng Des Sel. 2008 Nov;21(11):673-80. doi: 10.1093/protein/gzn048. Epub 
      2008 Sep 16.