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Literature summary extracted from

  • Shaya, D.; Hahn, B.; Park, N.Y.; Sim, J.; Kim, Y.S.; Cygler, M.
    Characterization of Chondroitin Sulfate Lyase ABC from Bacteroides thetaiotaomicron WAL2926 (2008), Biochemistry, 47, 6650-6661.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
4.2.2.21 expressed in Escherichia coli as a His-tagged fusion protein Bacteroides thetaiotaomicron

Protein Variants

EC Number Protein Variants Comment Organism
4.2.2.21 E628D mutant shows no activity Bacteroides thetaiotaomicron
4.2.2.21 E628Q Km (mM) (chondroitin sulfate A) 0.158, (chondroitin sulfate C) 0.035, kcat (1/sec) (chondroitin sulfate A) 10.38, (chondroitin sulfate B) 4.41, no activity toward dermatan sulfate Bacteroides thetaiotaomicron
4.2.2.21 H344D mutant shows no activity Bacteroides thetaiotaomicron
4.2.2.21 H344E mutant shows no activity Bacteroides thetaiotaomicron
4.2.2.21 H344N Km (mM) (chondroitin sulfate A) 0.039, (chondroitin sulfate C) 0.084, (dermatan sulfate) 0.09, kcat (1/sec) (chondroitin sulfate A) 42.2, (chondroitin sulfate B) 18.7, (dermatan sulfate) 9.06 Bacteroides thetaiotaomicron
4.2.2.21 H344Q Km (mM) (chondroitin sulfate A) 0.073, kcat (1/sec) (chondroitin sulfate A) 16.78, no activity toward dermatan sulfate or chondroitin sulfate C Bacteroides thetaiotaomicron
4.2.2.21 H345D/E mutant shows no activity Bacteroides thetaiotaomicron
4.2.2.21 H345N Km (mM) (chondroitin sulfate A) 0.061, (chondroitin sulfate C) 0.026, kcat (1/sec) (chondroitin sulfate A) 21, (chondroitin sulfate B) 6.6, no activity toward dermatan sulfate Bacteroides thetaiotaomicron
4.2.2.21 H345Q Km (mM) (chondroitin sulfate A) 0.045, (chondroitin sulfate C) 0.102, kcat (1/sec) (chondroitin sulfate A) 23.46, (chondroitin sulfate B) 10.85, no activity toward dermatan sulfate Bacteroides thetaiotaomicron
4.2.2.21 H453N Km (mM) (chondroitin sulfate A) 0.034, (chondroitin sulfate C) 0.012, kcat (1/sec) (chondroitin sulfate A) 13.15, (chondroitin sulfate B) 6.8, no activity toward dermatan sulfate Bacteroides thetaiotaomicron
4.2.2.21 H454/D/N/Q mutant shows no activity Bacteroides thetaiotaomicron
4.2.2.21 Y461A mutant shows no activity Bacteroides thetaiotaomicron

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
4.2.2.21 0.012
-
 chondroitin sulfate C mutant H453N Bacteroides thetaiotaomicron
4.2.2.21 0.026
-
 chondroitin sulfate C mutant H345N Bacteroides thetaiotaomicron
4.2.2.21 0.033
-
 chondroitin sulfate C wild-type Bacteroides thetaiotaomicron
4.2.2.21 0.034
-
 chondroitin sulfate A mutant H453N Bacteroides thetaiotaomicron
4.2.2.21 0.035
-
 chondroitin sulfate C mutant E628Q Bacteroides thetaiotaomicron
4.2.2.21 0.039
-
 chondroitin sulfate A mutant H344N Bacteroides thetaiotaomicron
4.2.2.21 0.045
-
 chondroitin sulfate A mutant H345Q Bacteroides thetaiotaomicron
4.2.2.21 0.061
-
 dermatan sulfate wild-type Bacteroides thetaiotaomicron
4.2.2.21 0.061
-
 chondroitin sulfate A mutant H345N Bacteroides thetaiotaomicron
4.2.2.21 0.067
-
 chondroitin sulfate A wild-type Bacteroides thetaiotaomicron
4.2.2.21 0.073
-
 chondroitin sulfate A mutant H344Q Bacteroides thetaiotaomicron
4.2.2.21 0.084
-
 chondroitin sulfate C mutant H344N Bacteroides thetaiotaomicron
4.2.2.21 0.09
-
 dermatan sulfate mutant H344N Bacteroides thetaiotaomicron
4.2.2.21 0.102
-
 chondroitin sulfate C mutant H345Q Bacteroides thetaiotaomicron
4.2.2.21 0.158
-
 chondroitin sulfate A mutant E628Q Bacteroides thetaiotaomicron

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
4.2.2.21 Ca2+ stimulates activity, particularly against dermatan sulfate Bacteroides thetaiotaomicron
4.2.2.21 Mg2+ stimulates activity, particularly against dermatan sulfate Bacteroides thetaiotaomicron
4.2.2.21 additional information for the monovalent ions tested, increasing the concentration of Na+, K+, or CS+ chloride from 0 to 0.1 M increases the activity against all substrates. Further increases in salt concentration reduces the activity dramatically, with 50% inhibition occurring at 0.15 M and a nearly complete inhibition at 0.4 M salt Bacteroides thetaiotaomicron

Organism

EC Number Organism UniProt Comment Textmining
4.2.2.21 Bacteroides thetaiotaomicron Q8A2I1
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
4.2.2.21 using a combination of DEAE-, Ni-NTA-, Mono-S HR 10/10, and HiLoad 16/60 Superdex 200-columns Bacteroides thetaiotaomicron

Reaction

EC Number Reaction Comment Organism Reaction ID
4.2.2.21 [GlcAbeta(1-3)GalNAc(4S)beta(1-4)]3 = [GlcAbeta(1-3)GalNAc(4S)beta(1-4)]2 + 2-(acetylamino)-2-deoxy-3-O-alpha-L-threo-hex-4-enopyranuronosyl-4-O-sulfo-beta-D-galactopyranose only disaccharide products and no tetrasaccharide for each of the substrates tested are found during the course of digestion, consistent with an exolytic mode of action for the enzyme Bacteroides thetaiotaomicron
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
4.2.2.21 chondroitin sulfate A bovine trachea, relative activity: 100% Bacteroides thetaiotaomicron ?
-
 ?
4.2.2.21 chondroitin sulfate C shark cartilage, relative activity: 61.1% Bacteroides thetaiotaomicron ?
-
 ?
4.2.2.21 chondroitin sulfate D skate cartilage, relative activity: 18.5% Bacteroides thetaiotaomicron ?
-
 ?
4.2.2.21 chondroitin sulfate E skate cartilage, relative activity: 36.7% Bacteroides thetaiotaomicron ?
-
 ?
4.2.2.21 dermatan sulfate porcine intestinal mucosa, relative activity: 9.1% Bacteroides thetaiotaomicron ?
-
 ?
4.2.2.21 high molecular weight hyaluronic acid D relative activity: 1.1% Bacteroides thetaiotaomicron ?
-
 ?
4.2.2.21 low molecular weight hyaluronic acid D relative activity: 1.1% Bacteroides thetaiotaomicron ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
4.2.2.21 BactnABC
-
 Bacteroides thetaiotaomicron
4.2.2.21 Chondroitin Sulfate Lyase ABC
-
 Bacteroides thetaiotaomicron

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
4.2.2.21 37
-
-
 Bacteroides thetaiotaomicron

Temperature Range [°C]

EC Number Temperature Minimum [°C] Temperature Maximum [°C] Comment Organism
4.2.2.21 25 40 no significant reduction in activity at temperatures in the range of 25-40 °C Bacteroides thetaiotaomicron

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
4.2.2.21 25 40 above 40°C the enzyme gets denaturated Bacteroides thetaiotaomicron

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
4.2.2.21 4.41
-
 chondroitin sulfate C mutant E628Q Bacteroides thetaiotaomicron
4.2.2.21 6.6
-
 chondroitin sulfate C mutant H345N Bacteroides thetaiotaomicron
4.2.2.21 6.8
-
 chondroitin sulfate C mutant H453N Bacteroides thetaiotaomicron
4.2.2.21 9.06
-
 dermatan sulfate mutant H344N Bacteroides thetaiotaomicron
4.2.2.21 10.38
-
 chondroitin sulfate A mutant E628Q Bacteroides thetaiotaomicron
4.2.2.21 10.85
-
 chondroitin sulfate C mutant H345Q Bacteroides thetaiotaomicron
4.2.2.21 13.15
-
 chondroitin sulfate A mutant H453N Bacteroides thetaiotaomicron
4.2.2.21 16.78
-
 chondroitin sulfate A mutant H344Q Bacteroides thetaiotaomicron
4.2.2.21 18.7
-
 chondroitin sulfate C mutant H344N Bacteroides thetaiotaomicron
4.2.2.21 21
-
 chondroitin sulfate A mutant H345N Bacteroides thetaiotaomicron
4.2.2.21 23.46
-
 chondroitin sulfate A mutant H345Q Bacteroides thetaiotaomicron
4.2.2.21 38.45
-
 dermatan sulfate wild-type Bacteroides thetaiotaomicron
4.2.2.21 42.2
-
 chondroitin sulfate A mutant H344N Bacteroides thetaiotaomicron
4.2.2.21 173
-
 chondroitin sulfate C wild-type Bacteroides thetaiotaomicron
4.2.2.21 263.2
-
 chondroitin sulfate A wild-type Bacteroides thetaiotaomicron

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
4.2.2.21 7.6
-
-
 Bacteroides thetaiotaomicron

pH Range

EC Number pH Minimum pH Maximum Comment Organism
4.2.2.21 7 8
-
 Bacteroides thetaiotaomicron