Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.2.1.62 extracted from

  • Durand, J.; Biarnes, X.; Watterlot, L.; Bonzom, C.; Borsenberger, V.; Planas, A.; Bozonnet, S.; O'Donohue, M.; Faure, R.
    A single point mutation alters the transglycosylation/Hydrolysis partition, significantly enhancing the synthetic capability of an endo-glycoceramidase (2016), ACS Catal., 6, 8264-8275 .
No PubMed abstract available
Search for more literature for 3.2.1.62

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Rhodococcus sp. M-777

Protein Variants

Protein Variants Comment Organism
D311Y the mutant enzyme is characterized by a lowered KM value for the hydrolysis of 2-chloro-4-nitrophenyl beta-cellobioside and increased transglycosylation when using aliphatic 1,3-diols or alcohols bearing a delta-hydroxy ketone function as acceptors. The transglycosylation/hydrolysis ratio in reactions catalyzed by the mutant is completely inversed and weak secondary hydrolysis is postponed, thus providing the basis for high transglycosylation yields (between 68 and 93%) Rhodococcus sp. M-777
D311Y the mutant shows increased catalytic efficiency for the hydrolysis of 2-chloro-4-nitrophenyl beta-cellobioside compared to the wild type enzyme. The transglycosylation ability of the mutant is improved in comparison to the wild type enzyme Rhodococcus sp. M-777

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
7
-
 2-chloro-4-nitrophenyl beta-cellobioside mutant enzyme D311Y, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777
22.4
-
 2-chloro-4-nitrophenyl beta-cellobioside wild type enzyme, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777
22.7
-
 2-chloro-4-nitrophenyl beta-cellobioside mutant enzyme D314Y, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777

Organism

Organism UniProt Comment Textmining
Rhodococcus sp. M-777 O33853
-
-

Purification (Commentary)

Purification (Comment) Organism
TALON metal affinity resin column chromatography Rhodococcus sp. M-777

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-chloro-4-nitrophenyl beta-cellobioside + H2O poorly recognized substrate Rhodococcus sp. M-777 2-chloro-4-nitrophenol + beta-cellobiose
-
 ir
alpha-cellobioside + 1-hydroxyoctan-4-one transglycosylation reaction Rhodococcus sp. M-777 4-oxooct-1-yl beta-D-glucopyranosyl(1->4)-beta-D-glucopyranoside
-
 r
alpha-cellobioside + 5-hydroxypentan-2-one transglycosylation reaction Rhodococcus sp. M-777 2-oxopent-5-yl beta-D-glucopyranosyl(1->4)-beta-D-glucopyranoside
-
 r
alpha-cellobioside + octane-1,3-diol transglycosylation reaction Rhodococcus sp. M-777 3-hydroxyoct-1-yl beta-D-glucopyranosyl(1->4)-beta-D-glucopyranoside + 1-(2-hydroxy)ethylhex-1-yl beta-D-glucopyranosyl(1->4)-beta-D-glucopyranoside
-
 r
alpha-cellobioside + pentane-1,3-diol transglycosylation reaction Rhodococcus sp. M-777 3-hydroxypent-1-yl beta-D-glucopyranosyl(1->4)-beta-D-glucopyranoside + 1-ethyl(3-hydroxy)prop-1-yl beta-D-glucopyranosyl(1->4)-beta-D-glucopyranoside
-
 r

Synonyms

Synonyms Comment Organism
EGC
-
 Rhodococcus sp. M-777
endo-glycoceramidase II
-
 Rhodococcus sp. M-777

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.9
-
 2-chloro-4-nitrophenyl beta-cellobioside mutant enzyme D311Y, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777
3.7
-
 2-chloro-4-nitrophenyl beta-cellobioside wild type enzyme, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777
5.4
-
 2-chloro-4-nitrophenyl beta-cellobioside mutant enzyme D314Y, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.17
-
 2-chloro-4-nitrophenyl beta-cellobioside wild type enzyme, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777
0.24
-
 2-chloro-4-nitrophenyl beta-cellobioside mutant enzyme D314Y, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777
0.42
-
 2-chloro-4-nitrophenyl beta-cellobioside mutant enzyme D311Y, hydrolysis reaction, at pH 4.0 and 37°C Rhodococcus sp. M-777