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Literature summary 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)

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

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.19 Q265K the mutant shows 15% cyclization (alpha-cyclodextrin-forming) and 236% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
2.4.1.19 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
2.4.1.19 Q265K/Y195S the mutant shows no cyclization (alpha-cyclodextrin-forming) and 226% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
2.4.1.19 Y195S the mutant shows no cyclization (alpha-cyclodextrin-forming) and 200% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
2.4.1.19 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
2.4.1.19 Y195S/Q265K compared with the wild type enzyme, the mutant has no cyclization activity and 498% hydrolysis and disproportionation activity Paenibacillus macerans
2.4.1.19 Y195S/Y260R compared with the wild type enzyme, the mutant has lower cyclization activity and higher hydrolysis and disproportionation activity Paenibacillus macerans
2.4.1.19 Y260R the mutant shows no cyclization (alpha-cyclodextrin-forming) and 226% hydrolysis (starch-degrading) activities compared to the wild type enzyme Paenibacillus macerans
2.4.1.19 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
2.4.1.19 Y260R/Q265K compared with the wild type enzyme, the mutant has 8% cyclization activity and 213% hydrolysis activity Paenibacillus macerans
2.4.1.19 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
2.4.1.19 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

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
2.4.1.19 0.47
-
 maltodextrin mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.48
-
 maltodextrin mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.5
-
 maltodextrin mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.51
-
 maltodextrin mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.52
-
 maltodextrin mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.53
-
 maltodextrin mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.62
-
 maltodextrin mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.64
-
 maltodextrin wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 38.32
-
 L-ascorbic acid wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 43.44
-
 L-ascorbic acid mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 44.32
-
 L-ascorbic acid mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 44.52
-
 L-ascorbic acid mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 47.21
-
 L-ascorbic acid mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 48.47
-
 L-ascorbic acid mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 48.84
-
 L-ascorbic acid mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 51.07
-
 L-ascorbic acid mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
2.4.1.19 75000
-
 x * 75000, SDS-PAGE Paenibacillus macerans

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.19 Paenibacillus macerans
-
-
-
2.4.1.19 Paenibacillus macerans JFB05-01
-
-
-

Purification (Commentary)

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

EC Number Synonyms Comment Organism
2.4.1.19 CGTase
-
 Paenibacillus macerans

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.19 36
-
-
 Paenibacillus macerans

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
2.4.1.19 40
-
 the half-life at 40°C is 7.6 h Paenibacillus macerans

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
2.4.1.19 0.000086
-
 L-ascorbic acid mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.000097
-
 L-ascorbic acid wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0001
-
 L-ascorbic acid mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.000105
-
 L-ascorbic acid mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.000106
-
 L-ascorbic acid mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.00011
-
 L-ascorbic acid mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.000114
-
 L-ascorbic acid mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.000133
-
 L-ascorbic acid mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0059
-
 maltodextrin wild type enzyme, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0067
-
 maltodextrin mutant enzyme Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0084
-
 maltodextrin mutant enzyme Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.009
-
 maltodextrin mutant enzyme Y260R, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0096
-
 maltodextrin mutant enzyme Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0098
-
 maltodextrin mutant enzyme Y260R/Y195S, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0104
-
 maltodextrin mutant enzyme Y260R/Q265K, at pH 5.5 and 37°C Paenibacillus macerans
2.4.1.19 0.0124
-
 maltodextrin mutant enzyme Y260R/Q265K/Y195S, at pH 5.5 and 37°C Paenibacillus macerans

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.19 6.5
-
-
 Paenibacillus macerans

pH Stability

EC Number pH Stability pH Stability Maximum Comment Organism
2.4.1.19 5.5
-
 the half-life at pH 5.5 is 29.8 h Paenibacillus macerans