RebH

Enzyme Information
reference: 10.1002/prot.21627
host: E. coli strain BL21(DE3)
test: 303 K in 100 μL total volume 20 mM potassium phosphate buffer (pH 8.0) containing 0.6 mM L-Trp, 0.2 mg/mL RebH, 50 μM FAD and 100 mM halide (NaCl or NaBr) and an in situ flavin reduction system

reference: 10.1002/cbic.202100210
host: E. coli
test: mutagenesis

Gene Data
phmm effect mutation gene_name reference
104 flipped peptide carbonyl upon Trp binding G112 RebH DOI: 10.1002/prot.21627
105 displace side chain upon Trp binding L113 RebH DOI: 10.1002/prot.21627
- forms a salt bridge to stabilize the dimer interface R387-E432 RebH DOI: 10.1002/prot.21627
- has parental 3-LSR;>=2-fold higher activity over parental 3-LSRM; for indole bromination Y455C RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR;>=2-fold higher activity over parental 3-LSRM; for indole bromination F465N RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR;>=2-fold higher activity over parental 3-LSRM; for indole bromination W466I RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR;>=2-fold higher activity over parental 3-LSRM; for indole bromination W466Y RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR;>=2-fold higher activity over parental 3-LSRM; for indole bromination N470S RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR;>=2-fold higher activity over parental 3-LSRM; for indole bromination N470K RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR; >=2-fold higher activity over parental 3-LSRM; for indole bromination F465K RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR; 4.5-fold improved conversion of brominated indole Y455C/F465K RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR; 4.2-fold improved conversion of brominated indole Y455W/N470S RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR; 80% activity loss F465K/N470K RebH DOI: 10.1002/cbic.202100210
- has parental 3-LSR; 80% activity loss F465N/N570K RebH DOI: 10.1002/cbic.202100210
- high halogenation conversion S130L/Q494R RebH DOI: 10.1002/cbic.201300780
- high halogenation conversion S130L/N166S/Q494R RebH DOI: 10.1002/cbic.201300780
- 20-fold improvement in conversion compared to WT S2P/M71V/K145M RebH DOI: 10.1002/cbic.201300780
- has parental 1-PVM; 2.5-fold increase in halogenation conversion on Trp E461G RebH DOI: 10.1002/cbic.201300780
- has parental 1-PVM; 2.5-fold increase in halogenation conversion on Trp E423D RebH DOI: 10.1002/cbic.201300780
- increased stability compared to WT S2P RebH DOI: 10.1002/cbic.201300780
- has parental 3-LSR; >=2-fold higher activity over parental 3-LSRM; for indole bromination; produces halogenated alkaloid 12-Cl-19,20-dihydroakuammicine; can halogenate tryptamine Y455W RebH DOI: 10.1002/cbic.202100210; 10.1021/ja2089348
79 active site; located directlz between the flavin and tryptophan binding sites K79 RebH DOI: 10.1021/bi0621213
- interacts with its hydroxyl with K79 S347 RebH DOI: 10.1021/bi0621213
- affected in flavin oxidation and overall halogenation activity K79A RebH DOI: 10.1021/bi0621213
- affected in flavin oxidation and overall halogenation activity K79M RebH DOI: 10.1021/bi0621213
347 proposed to be involved in stabilization of the protonated Cl-Trp intermediate E357 RebH DOI: 10.1021/bi0621213