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Literature summary for 4.2.2.21 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)

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

Protein Variants

Protein Variants Comment Organism
E628D mutant shows no activity Bacteroides thetaiotaomicron
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
H344D mutant shows no activity Bacteroides thetaiotaomicron
H344E mutant shows no activity Bacteroides thetaiotaomicron
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
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
H345D/E mutant shows no activity Bacteroides thetaiotaomicron
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
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
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
H454/D/N/Q mutant shows no activity Bacteroides thetaiotaomicron
Y461A mutant shows no activity Bacteroides thetaiotaomicron

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ stimulates activity, particularly against dermatan sulfate Bacteroides thetaiotaomicron
Mg2+ stimulates activity, particularly against dermatan sulfate Bacteroides thetaiotaomicron
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

Organism UniProt Comment Textmining
Bacteroides thetaiotaomicron Q8A2I1
-
-

Purification (Commentary)

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

Reaction

Reaction Comment Organism Reaction ID
[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

Substrates and Products (Substrate)

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

Synonyms

Synonyms Comment Organism
BactnABC
-
Bacteroides thetaiotaomicron
Chondroitin Sulfate Lyase ABC
-
Bacteroides thetaiotaomicron

Temperature Optimum [°C]

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

Temperature Range [°C]

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

Temperature Stability [°C]

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

Turnover Number [1/s]

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

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
-
Bacteroides thetaiotaomicron

pH Range

pH Minimum pH Maximum Comment Organism
7 8
-
Bacteroides thetaiotaomicron