Skip to main content
. Author manuscript; available in PMC: 2017 Dec 6.
Published in final edited form as: Adv Protein Chem Struct Biol. 2017 Jun 5;109:223–264. doi: 10.1016/bs.apcsb.2017.04.008

Table 2.

Activity of Sortase Enzymes In Vitro

Sortase Primary Substrate (Km mM) Secondary
Substrate (Km µM)
Cleavage kcat (s−1) Transpeptidation
kcat (s−1)
Class A
SaSrtAΔ24 (Frankel, Kruger, Robinson, Kelleher, & McCafferty, 2005) Abz-LPETG-Dap(Dnp) (7.33 ± 1.01) Gly5 (196 ± 64) 0.086 ± 0.015 0.28 ± 0.02
SaSrtAΔ24 (Kruger, Dostal, & McCafferty, 2004) Abz-LPETG-Dap(Dnp) (5.5 mM) Gly5 (140) NR 0.27
SaSrtAΔ24 (Bentley, Lamb, & McCafferty, 2008) Abz-LPETGG-Dap(Dnp) (8.76 ± 0.78)a Gly5 (NR) NR 1.10 ± 0.06a
SaSrtAΔ59 (Chen, Dorr, & Liu, 2011) Abz-LPETGK-(Dnp) (7.6 ± 0.5) Gly3 (140 ± 30) NR 1.5 ± 0.2
SaSrtAΔ59 Evolved tetramutant (Chen et al., 2011) Abz-LPETGK-(Dnp) (0.17 ± 0.03) Gly3 (4800 ± 700) NR 4.8 ± 0.8
SpySrtAΔ81 (Race et al., 2009) Abz-LPETGG-Dap(Dnp) (0.83 ± 0.11) Ala2 (NR) NR 0.0136 ± 0.0011
BaSrtAΔ56 (Weiner, Robson, Marohn, & Clubb, 2010) Abz-LPETG-Dap(Dnp) (0.038 ± 4)b m-DAPc 0.0004 ± 0.0001b c
BaSrtAΔ56 (Chan et al., 2015) Abz-LPETG-Dap(Dnp) (0.306 ± 0.023) m-DAPc 3.6 ± 0.2 × 10−5 c
BaSrtAΔ64 (Chan et al., 2015) Abz-LPETG-Dap(Dnp) (0.173 ± 0.011) m-DAPc 5.7 ± 0.2 × 10−5 c
SmutSrtAΔ40 (Wallock-Richards et al., 2015) Dabcyl-QALPETGEE-Edans (0.0904 ± 0.0047)b NR Yes NR
Class B
SaSrtBΔ21 (Bentley, Gaweska, Kielec, & McCafferty, 2007) Abz-KVENPQTNAGT-Dap(DNP) (7.8 ± 2) Gly5 (NR) NR 5.4 ± 0.5 × 10−4
SaSrtBΔ31 (Jacobitz et al., 2014) SNKDKVENPQTNAGT (1.8) Gly5 (NR) NR 1.010−4
BaSrtBΔ37 (Maresso, Chapa, & Schneewind, 2006) Abz -KTDNPKTGDEA-Dap(DNP) NR Yes NR
CdSrtBΔ26 (van Leeuwen et al., 2014) KIVKSPKTGDETQLMK KPPVPPKTGDSTTIGK Gly4/5 or Ala-d-Glu-DAP NR NR
Class C
SpnSrtC117–228 (Manzano et al., 2008) IPQTG in RrgB30–633 YPKN in RrgB30–633 No Yes
SagSrtC143–254 from PI-2a (Cozzi et al., 2011) Dabcyl-KKVTIPQTGGIGT-Edans (0.0138) NR Yes NR
SagSrtC142–305 from PI-1 (Cozzi et al., 2012) Dabcyl-RPPGVFPKTGGIG-Edans (0.0 1358 ± 0.00063) NR 1.16 ± 0.044 × 10−3 NR
Dabcyl-RPSIPNTGGIG-Edans (0.03100 ± 0.00462) NR 1.77 ± 0.101 × 10−3 NR
Dabcyl-RGGLIPKTGEQQ-Edans (0.01639 ± 0.00250) NR 0.77 ± 0.038 × 10−3 NR
SagSrtC242–283 from PI-1 (Cozzi et al., 2012) Dabcyl-RPPGVFPKTGGIG-Edans (0.006385 ± 0.00142) NR 1.04 ± 0.058 × 10−3 NR
Dabcyl-RGGLIPKTGEQQ-Edans (0.02733 ± 0.00435) NR 4.36 ± 0.256 × 10−4 NR
Dabcyl-RPSIPNTGGIG-Edans (0.05715 ± 0.00354) NR 5.56 ± 0.174 × 10−3 NR
Class D
BaSrtDΔ55 (Robson, Jacobitz, Phillips, & Clubb, 2012) VQGEKLPNTASNN m-DAP (NR) Yes NR
BaSrtDΔ55 (Marraffini & Schneewind, 2006) Abz-GEKLPNTASNN-Dnp m-DAP (NR) Yes NR
CpSrtDΔ23–187 (Suryadinata, Seabrook, Adams, Nuttall, & Peat, 2015) Aβ1–16-LPQTGS NR Yes NR
a

Values reported from fluorescence assay and subject to inner filter effect and are likely underestimates of true parameters.

b

These values calculated assuming a hydrolytic shunt mechanism.

c

The enzyme reportedly does not perform this reaction in vitro.

Sorting signals for all substrates are highlighted in bold.

Errors are reported where published.

“Yes” indicates the reaction was performed in vitro but kinetics parameters were not reported.

NR, not reported.

m-DAP, mesodiaminopimelic acid.