Chem Int Ed Engl 59:11142–11149. https://
doi.org/10.1002/anie.202002899
25. Spinck M, Ecke M, Sievers S, Neumann H
(2018) Highly sensitive lysine deacetylase
assay based on acetylated firefly luciferase. Biochemistry 57:3552–3555. https://doi.org/
10.1021/acs.biochem.8b00483
26. Neumann H, Peak-Chew SY, Chin JW (2008)
Genetically encoding N(epsilon)-acetyllysine in
recombinant proteins. Nat Chem Biol
4:232–234.
https://doi.org/10.1038/
nchembio.73
27. Stemmer WP, Morris SK (1992) Enzymatic
inverse PCR: a restriction site independent,
single-fragment method for high-efficiency,
site-directed mutagenesis. BioTechniques
13:214–220
28. Zhao K, Chai X, Marmorstein R (2004) Structure and substrate binding properties of cobB,
a Sir2 homolog protein deacetylase from
Escherichia coli. J Mol Biol 337:731–741.
https://doi.org/10.1016/j.jmb.2004.01.060
29. Hoff KG, Avalos JL, Sens K, Wolberger C
(2006) Insights into the Sirtuin mechanism
from ternary complexes containing NAD+
and acetylated peptide. Structure 14
(8):1231–1240
30. Pyne ME, Moo-Young M, Chung DA, Chou
CP (2015) Coupling the CRISPR/Cas9 system with lambda red Recombineering enables
simplified chromosomal gene replacement in
Escherichia coli. Appl Environ Microbiol
81:5103–5114.
https://doi.org/10.1128/
aem.01248-15
31. Appleby TC, Kinsland C, Begley TP, Ealick SE
(2000) The crystal structure and mechanism of
orotidine 5
0 -monophosphate decarboxylase.
Proc Natl Acad Sci U S A 97:2005–2010.
https://doi.org/10.1073/pnas.259441296
32. Boeke JD, Trueheart J, Natsoulis G, Fink GR
(1987) 5-Fluoroorotic acid as a selective agent
in yeast molecular genetics. Methods Enzymol
154:164–175
A Directed Evolution System for Lysine Deacetylases
337
doi.org/10.1002/anie.202002899
25. Spinck M, Ecke M, Sievers S, Neumann H
(2018) Highly sensitive lysine deacetylase
assay based on acetylated firefly luciferase. Biochemistry 57:3552–3555. https://doi.org/
10.1021/acs.biochem.8b00483
26. Neumann H, Peak-Chew SY, Chin JW (2008)
Genetically encoding N(epsilon)-acetyllysine in
recombinant proteins. Nat Chem Biol
4:232–234.
https://doi.org/10.1038/
nchembio.73
27. Stemmer WP, Morris SK (1992) Enzymatic
inverse PCR: a restriction site independent,
single-fragment method for high-efficiency,
site-directed mutagenesis. BioTechniques
13:214–220
28. Zhao K, Chai X, Marmorstein R (2004) Structure and substrate binding properties of cobB,
a Sir2 homolog protein deacetylase from
Escherichia coli. J Mol Biol 337:731–741.
https://doi.org/10.1016/j.jmb.2004.01.060
29. Hoff KG, Avalos JL, Sens K, Wolberger C
(2006) Insights into the Sirtuin mechanism
from ternary complexes containing NAD+
and acetylated peptide. Structure 14
(8):1231–1240
30. Pyne ME, Moo-Young M, Chung DA, Chou
CP (2015) Coupling the CRISPR/Cas9 system with lambda red Recombineering enables
simplified chromosomal gene replacement in
Escherichia coli. Appl Environ Microbiol
81:5103–5114.
https://doi.org/10.1128/
aem.01248-15
31. Appleby TC, Kinsland C, Begley TP, Ealick SE
(2000) The crystal structure and mechanism of
orotidine 5
0 -monophosphate decarboxylase.
Proc Natl Acad Sci U S A 97:2005–2010.
https://doi.org/10.1073/pnas.259441296
32. Boeke JD, Trueheart J, Natsoulis G, Fink GR
(1987) 5-Fluoroorotic acid as a selective agent
in yeast molecular genetics. Methods Enzymol
154:164–175
A Directed Evolution System for Lysine Deacetylases
337