Cross‐linked enzyme aggregates of recombinant cyclodextrin glycosyltransferase for high‐purity β‐cyclodextrin production

2019 ◽  
Vol 94 (5) ◽  
pp. 1528-1533 ◽  
Author(s):  
Jianguo Zhang ◽  
Mengla Li ◽  
Hongli Mao
Keyword(s):  
High Purity ◽  
Cyclodextrin Glycosyltransferase ◽  
Cyclodextrin Production

Biochemical properties and cyclodextrin production profiles of isoforms of cyclodextrin glycosyltransferase

2010 ◽  
Vol 70 (3-4) ◽  
pp. 377-383 ◽  
Author(s):  
Wanchai Yenpetch ◽  
Kanoktip Packdibamrung ◽  
Wolfgang Zimmermann ◽  
Piamsook Pongsawasdi
Keyword(s):  
Biochemical Properties ◽  
Cyclodextrin Glycosyltransferase ◽  
Cyclodextrin Production

Rational design of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 to increase α-cyclodextrin production 1 1Edited by G. Von Heijne

2000 ◽  
Vol 296 (4) ◽  
pp. 1027-1038 ◽  
Author(s):  
Bart A van der Veen ◽  
Joost C.M Uitdehaag ◽  
Dirk Penninga ◽  
Gert-Jan W.M van Alebeek ◽  
Loraine M Smith ◽  
...  
Keyword(s):  
Rational Design ◽  
Bacillus Circulans ◽  
Cyclodextrin Glycosyltransferase ◽  
Cyclodextrin Production

scholarly journals Enhancing the α-Cyclodextrin Specificity of Cyclodextrin Glycosyltransferase from Paenibacillus macerans by Mutagenesis Masking Subsite −7

2016 ◽  
Vol 82 (8) ◽  
pp. 2247-2255 ◽  
Author(s):  
Lei Wang ◽  
Xuguo Duan ◽  
Jing Wu
Keyword(s):  
Active Site ◽  
Wild Type ◽  
Cyclodextrin Glycosyltransferase ◽  
Wild Type Enzyme ◽  
Widespread Application ◽  
Content Type ◽  
Hydrogen Bonding Interactions ◽  
Paenibacillus Macerans ◽  
Site Blocking ◽  
Cyclodextrin Production

ABSTRACTCyclodextrin glycosyltransferases (CGTases) (EC 2.4.1.19) catalyze the conversion of starch or starch derivates into mixtures of α-, β-, and γ-cyclodextrins. Because time-consuming and expensive purification procedures hinder the widespread application of single-ingredient cyclodextrins, enzymes with enhanced specificity are needed. In this study, we tested the hypothesis that the α-cyclodextrin selectivity ofPaenibacillus maceransα-CGTase could be augmented by masking subsite −7 of the active site, blocking the formation of larger cyclodextrins, particularly β-cyclodextrin. Five single mutants and three double mutants designed to remove hydrogen-bonding interactions between the enzyme and substrate at subsite −7 were constructed and characterized in detail. Although the rates of α-cyclodextrin formation varied only modestly, the rate of β-cyclodextrin formation decreased dramatically in these mutants. The increase in α-cyclodextrin selectivity was directly proportional to the increase in the ratio of theirkcatvalues for α- and β-cyclodextrin formation. The R146A/D147P and R146P/D147A double mutants exhibited ratios of α-cyclodextrin to total cyclodextrin production of 75.1% and 76.1%, approximately one-fifth greater than that of the wild-type enzyme (63.2%), without loss of thermostability. Thus, these double mutants may be more suitable for the industrial production of α-cyclodextrin than the wild-type enzyme. The production of β-cyclodextrin by these mutants was almost identical to their production of γ-cyclodextrin, which was unaffected by the mutations in subsite −7, suggesting that subsite −7 was effectively blocked by these mutations. Further increases in α-cyclodextrin selectivity will require identification of the mechanism or mechanisms by which these small quantities of larger cyclodextrins are formed.

scholarly journals Substrate binding to a cyclodextrin glycosyltransferase and mutations increasing the gamma-cyclodextrin production

1998 ◽  
Vol 255 (3) ◽  
pp. 710-717 ◽  
Author(s):  
Goetz Parsiegla ◽  
Andreas K. Schmidt ◽  
Georg E. Schulz
Keyword(s):  
Substrate Binding ◽  
Cyclodextrin Glycosyltransferase ◽  
Cyclodextrin Production ◽  
Gamma Cyclodextrin

Mutations in Cyclodextrin Glycosyltransferase fromBacillus circulansEnhance β-Cyclization Activity and β-Cyclodextrin Production

2014 ◽  
Vol 62 (46) ◽  
pp. 11209-11214 ◽  
Author(s):  
Min Huang ◽  
Caiming Li ◽  
Zhengbiao Gu ◽  
Li Cheng ◽  
Yan Hong ◽  
...  
Keyword(s):  
Cyclodextrin Glycosyltransferase ◽  
Cyclodextrin Production

scholarly journals Cyclodextrin production by Bacillus lehensis isolated from cassava starch: Characterisation of a novel enzyme

2014 ◽  
Vol 32 (No. 1) ◽  
pp. 48-53 ◽  
Author(s):  
K.C. Blanco ◽  
F.F. de Moraes ◽  
N.S. Bernardi ◽  
M.H.P.B. Vettori ◽  
R. Monti ◽  
...  
Keyword(s):  
Affinity Chromatography ◽  
Maximum Velocity ◽  
Temperature Stability ◽  
Cassava Starch ◽  
Optimum Temperature ◽  
Cyclodextrin Glycosyltransferase ◽  
Sds Page ◽  
Optimum Ph ◽  
Menten Constant ◽  
Cyclodextrin Production

The properties of a previously unknown enzyme, denominated cyclodextrin glycosyltransferase, produced from Bacillus lehensis, were evaluated using affinity chromatography for protein purification. Enzyme characteristics (optimum pH and temperature; pH and temperature stability), the influence of substances on the enzyme activity, enzyme kinetics, and cyclodextrin production were analysed. Cyclodextrin glycosyltransferase was purified up to 320.74-fold by affinity chromatography using β-cyclodextrin as the binder and it exhibited 8.71% activity recovery. This enzyme is a monomer with a molecular weight of 81.27 kDa, as estimated by SDS-PAGE. Optimum temperature and pH for cyclodextrin glycosyltransferase were 55°C and 8.0, respectively. The Michaelis-Menten constant was 8.62 g/l during maximum velocity of 0.858 g/l.h.  

scholarly journals Production and characterization of cyclodextrin glycosyltransferase from Bacillus.sp isolated from Cuban Soil.

2014 ◽  
Author(s):  
Héctor Luis. Ramirez Pérez
Keyword(s):  
Kinetic Parameters ◽  
Initial Rate ◽  
Cyclodextrin Glycosyltransferase ◽  
Product Concentration ◽  
Wide Ph Range ◽  
Significant Yield ◽  
Influence Of Substrate ◽  
Ph Range ◽  
Cyclodextrin Production

A cyclodextrin glycosyltransferase (CGTase) from an alkaliphilic Bacillus.sp strain, isolated from Cuba soil, was purified with Sephadex G-50 with a yield of 66.5 %. The CGTase was stable over a very wide pH range, 6.0 –10, at 25°C and was most active at pH 7.5. The enzyme exhibited an optimum temperature of 60°C and was stable to 50°C for at least 8 h. The T50 value – defined as the temperature at which 50% of the initial activity was retained– was 63 °C in this enzyme . The influence of substrate or product concentration on the initial rate of CD production was studied and the kinetic parameters were determined. The analysis of kinetic parameters Km and Vmax was obtained by the action of CGTase on the starch of corn with respect to β-CD and the values were 4.1 g/L and 5,2 μM β-CD/min ml respectively.. The purified CGTase from Bacillus.sp could be used for an efficient cyclodextrin production which significant yield of γ-cyclodextrins.

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