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

BRENDA support

Literature summary for 2.4.1.19 extracted from

  • Han, R.; Liu, L.; Shin, H.; Chen, R.; Li, J.; Du, G.; Chen, J.
    Systems engineering of tyrosine 195, tyrosine 260, and glutamine 265 in cyclodextrin glycosyltransferase from Paenibacillus macerans to enhance maltodextrin specificity for 2-O-D-glucopyranosyl-L-ascorbic acid synthesis (2013), Appl. Environ. Microbiol., 79, 672-677.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Paenibacillus macerans

Protein Variants

Protein Variants Comment Organism
Q265K the mutant shows 15% cyclization (alpha-cyclodextrin-forming) and 236% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
Q265K the mutant with enhanced maltodextrin specificity produces higher 2-O-D-glucopyranosyl-L-ascorbic acid yields than the wild type enzyme. Compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity Paenibacillus macerans
Q265K/Y195S the mutant shows no cyclization (alpha-cyclodextrin-forming) and 226% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
Y195S the mutant shows no cyclization (alpha-cyclodextrin-forming) and 200% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
Y195S the mutant with enhanced maltodextrin specificity produces higher 2-O-D-glucopyranosyl-L-ascorbic acid yields than the wild type enzyme. Compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity Paenibacillus macerans
Y195S/Q265K compared with the wild type enzyme, the mutant has no cyclization activity and 498% hydrolysis and disproportionation activity Paenibacillus macerans
Y195S/Y260R compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity Paenibacillus macerans
Y260R the mutant shows no cyclization (alpha-cyclodextrin-forming) and 226% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
Y260R the mutant with enhanced maltodextrin specificity produces higher 2-O-D-glucopyranosyl-L-ascorbic acid yields than the wild type enzyme. Compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity Paenibacillus macerans
Y260R/Q265K compared with the wild type enzyme, the mutant has 8% cyclization activity and 213% hydrolysis activity Paenibacillus macerans
Y260R/Q265K/Y195S the mutant shows 12% cyclization (alpha-cyclodextrin-forming) and 557% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
Y260R/Y195S the mutant shows no cyclization (alpha-cyclodextrin-forming) and 492% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans

Inhibitors

Inhibitors Comment Organism Structure
additional information not inhibited by L-ascorbic acid Paenibacillus macerans

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.47
-
maltodextrin mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.48
-
maltodextrin mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
0.5
-
maltodextrin mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.51
-
maltodextrin mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
0.52
-
maltodextrin mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
0.53
-
maltodextrin mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.62
-
maltodextrin mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.64
-
maltodextrin wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
38.32
-
L-ascorbic acid wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
43.44
-
L-ascorbic acid mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
44.32
-
L-ascorbic acid mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
44.52
-
L-ascorbic acid mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
47.21
-
L-ascorbic acid mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
48.47
-
L-ascorbic acid mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
48.84
-
L-ascorbic acid mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
51.07
-
L-ascorbic acid mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
75000
-
x * 75000, SDS-PAGE Paenibacillus macerans

Organism

Organism UniProt Comment Textmining
Paenibacillus macerans
-
-
-
Paenibacillus macerans JFB05-01
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography Paenibacillus macerans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
maltodextrin + L-ascorbic acid
-
Paenibacillus macerans 2-O-D-glucopyranosyl-L-ascorbic acid + ?
-
?
maltodextrin + L-ascorbic acid
-
Paenibacillus macerans JFB05-01 2-O-D-glucopyranosyl-L-ascorbic acid + ?
-
?

Subunits

Subunits Comment Organism
? x * 75000, SDS-PAGE Paenibacillus macerans

Synonyms

Synonyms Comment Organism
CGTase
-
Paenibacillus macerans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
36
-
-
Paenibacillus macerans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
40
-
the half-life at 40°C is 7.6 h Paenibacillus macerans

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.000086
-
L-ascorbic acid mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.000097
-
L-ascorbic acid wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
0.0001
-
L-ascorbic acid mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
0.000105
-
L-ascorbic acid mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
0.000106
-
L-ascorbic acid mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.00011
-
L-ascorbic acid mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
0.000114
-
L-ascorbic acid mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.000133
-
L-ascorbic acid mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.0059
-
maltodextrin wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
0.0067
-
maltodextrin mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.0084
-
maltodextrin mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
0.009
-
maltodextrin mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
0.0096
-
maltodextrin mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.0098
-
maltodextrin mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
0.0104
-
maltodextrin mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
0.0124
-
maltodextrin mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6.5
-
-
Paenibacillus macerans

pH Stability

pH Stability pH Stability Maximum Comment Organism
5.5
-
the half-life at pH 5.5 is 29.8 h Paenibacillus macerans