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Table 1 Comparison of different aspects concerning the production and selected properties of cholesterol oxidases

From: Cloning, expression and biochemical characterization of the cholesterol oxidase CgChoA from Chryseobacterium gleum

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)

[6, 24]

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

[21, 26]

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)

-

[10, 27, 28]

  1. Data taken from the Brenda database (http://www.brenda-enzymes.org/), RCSB Protein data bank (pdb) and indicated references (refs.).
  2. avector, host, expression strain, cultivation medium, cultivation conditions.
  3. bAbbreviations: ABTS 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), BSA bovine serum albumin, HRP horseradish peroxidase, IMAC Immobilized metal ion affinity chromatography, SEC size-exclusion chromatography.