Source (UniProt ID) | MW (kDa) | Expression systema | Purification stepsb | Activity asssayb | Specific activity on cholesterol (U/mg) | Refs. |
---|---|---|---|---|---|---|
Brevibacterium sterolicum (Q7SID9) | 61.52 | pET24b(+), E. coli BL21(DE3) pLysS, TB + 20 mL/L glycerol, 25/ 30°C | HiTrap (IMAC) | Potassium phosphate buffer pH 7.5 at 25°C (propan-2-ol and Thesit) monitoring H2O2 production using HRP and o-dianisidine | 7 (crude extract) | |
Brevibacterium sterolicum nov. sp. ATCC21387 | 46.50 | pUC19, E. coli MM294, LB, 30°C | Ammonium sulfate precipitation, DEAE-cellulose, Superose, hydroxyapatite | Sodium phosphate buffer pH 7.0, 37°C (Triton X-100), monitoring H2O2 production coupled to 4-aminoantipyrine and phenol via HRP | 55.2 | [25] |
Brevibacterium sterolicum nov. sp. ATCC21387 (Q2I2N2) | 59.07 | pET28a(+), E. coli BL21(DE3), LB, 28°C | Affinity chromato-graphy, (riboflavin bound to, Sepharose 4B) | Quantifying H2O2 by coupling to HRP reaction with aminopyrine, 37°C | 15.5 | [20] |
(3.7, with His tag) | [22] | |||||
Chromobacterium sp. DS-1 (B5MGF) | 63.00 | pET-21d(+), E. coli Rosetta, LB, 30°C | Heat purification at 70°C for 30 min, DEAE-cellulose DE52 | Sodium potassium phosphate buffer pH7.0, 30°C (sodium cholate, Triton X-100), 4-aminoantipyrine, phenol, HRP | 16.7 | |
Chryseobacterium gleum (D7VYA1) | 59.00 | pQE30 (pRARE2), E. coli JM109, modified M9, 16°C | HisTrap FF (IMAC), SEC on Superdex 200 pg | HPLC assay and coupled enzyme assay MOPS buffer, pH 6.75, 37°C, HRP and ABTS (Triton X-100 and taurocholic acid) | 15.5 | this study |
Streptomyces sp. SA-COO (P12676) | 58.99 | pUC19, E. coli BL21(DE3)plysS, 2YT, 28°C | Butyl-Sepharose column chromatography, DEAE-cellulose column chromatography | Formation rate of H2O2 was monitored in a coupled assay with HRP and ABTS, 37°C (Triton X-100, BSA) | - |