SalL

Enzyme Information
reference: 10.1038/nchembio.2007.56
host: E. coli BL21(DE3)
test: 1-300 nM SalL with 0.5–500 μM SAM and 10–500 mM KX (X = F, Cl, Br, I) or (ii) 0.5–100 mM L-methionine and 0.3–100 μM 5′-XDA (X = F, Cl, Br, I) in 50 mM phosphate buffer (pH 7.9) at 37 °C
producer: Salinospora tropica CNB-440
phmm-model: SAM_chlorinase.hmm

Gene Data
effect mutation gene_name reference pHMM_position
Y70T reduces activity by two magnitude; Y70T G131S 4-5 times increased activity (active site free of water) Y70 SalL DOI: 10.1021/acs.jafc.3c08185
5′-ClDA connected to monomer B Asn188 SalL DOI: 10.1038/nchembio.2007.56
no fluorination (previously Ser158 was hypothesized to be important for fluoride binding) G131S SalL DOI: 10.1021/acs.jafc.3c08185
fluorination V12L/Y70T/W129F/G131S SalL DOI: 10.1021/acs.jafc.3c08185
fluorination Y70T/W129F/G131S SalL DOI: 10.1021/acs.jafc.3c08185
increased 5'-FDA production; highest fluorination activity among the mutants Y70T/W129F SalL DOI: 10.1021/acs.jafc.3c08185
connected to 2′-OH and 3′-OH of ribose Asp11 SalL DOI: 10.1038/nchembio.2007.56
None F8 SalL DOI: 10.1021/acs.jafc.3c08185
None V12 SalL DOI: 10.1021/acs.jafc.3c08185
None F45 SalL DOI: 10.1021/acs.jafc.3c08185
in the double mutant Ser158 disrupts the H-bonding network with Thr75 Thr75 SalL DOI: 10.1021/acs.jafc.3c08185
W129F reduces activity W129 SalL DOI: 10.1021/acs.jafc.3c08185
5′-ClDA is hydrogen bonded to monomer A; G131S detrimental to halogenation; amide backbone connected to Cl; Y70T G131S 4-5 times increased activity (active site free of water) Gly131 SalL DOI: 10.1038/nchembio.2007.56
L-methionine forms hydrogen bonds to only monomer B Ser142 SalL DOI: 10.1038/nchembio.2007.56
L-methionine forms hydrogen bonds to only monomer B Arg243 SalL DOI: 10.1038/nchembio.2007.56
None A18S/Y70T SalL DOI: 10.1021/acs.jafc.3c08185
None F8Y/Y70T SalL DOI: 10.1021/acs.jafc.3c08185
None Y70T SalL DOI: 10.1021/acs.jafc.3c08185
None Y70G SalL DOI: 10.1021/acs.jafc.3c08185
None Y70P SalL DOI: 10.1021/acs.jafc.3c08185
None Y70W SalL DOI: 10.1021/acs.jafc.3c08185
fluorination Y70L SalL DOI: 10.1021/acs.jafc.3c08185
fluorination Y70I SalL DOI: 10.1021/acs.jafc.3c08185
fluorination Y70M SalL DOI: 10.1021/acs.jafc.3c08185
fluorination Y70T SalL DOI: 10.1021/acs.jafc.3c08185
fluorination Y70S SalL DOI: 10.1021/acs.jafc.3c08185
fluorination Y70C SalL DOI: 10.1021/acs.jafc.3c08185
fluorintation Y70A SalL DOI: 10.1021/acs.jafc.3c08185
maintained the majority of the chlorination activity F45A SalL DOI: 10.1021/acs.jafc.3c08185
maintained the majority of the chlorination activity, fluorination as well V12A SalL DOI: 10.1021/acs.jafc.3c08185
lower fluorination activity compared to Y70T V12S/Y70T SalL DOI: 10.1021/acs.jafc.3c08185
lower fluorination activity compared to Y70T V12D/Y70T SalL DOI: 10.1021/acs.jafc.3c08185
lower fluorination activity compared to Y70T V12N/Y70T SalL DOI: 10.1021/acs.jafc.3c08185