Rational design of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 to increase alpha-cyclodextrin production.

@article{vanderVeen2000RationalDO,
  title={Rational design of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 to increase alpha-cyclodextrin production.},
  author={Bart A. van der Veen and Joost C. M. Uitdehaag and Dirk Penninga and G J van Alebeek and Landon M. Smith and Bauke W. Dijkstra and Lubbert Dijkhuizen},
  journal={Journal of molecular biology},
  year={2000},
  volume={296 4},
  pages={
          1027-38
        }
}
Cyclodextrin glycosyltransferases (CGTase) (EC 2.4.1.19) are extracellular bacterial enzymes that generate cyclodextrins from starch. All known CGTases produce mixtures of alpha, beta, and gamma-cyclodextrins. A maltononaose inhibitor bound to the active site of the CGTase from Bacillus circulans strain 251 revealed sugar binding subsites, distant from the catalytic residues, which have been proposed to be involved in the cyclodextrin size specificity of these enzymes. To probe the importance… 
View on PubMed
rug.nl
91 Citations
Highly Influential Citations
7
Background Citations
20
Methods Citations
6
Results Citations
3

Figures and Tables from this paper

91 Citations

Mutations at subsite −3 in cyclodextrin glycosyltransferase from Paenibacillus macerans enhancing α-cyclodextrin specificity

The D372K and Y89R single and double mutants were much more suitable for the industrial production of α-cyclodextrin than the wild-type enzyme, which might be a result of stabilizing the bent conformation of the intermediate in the cyclization reaction.

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

The hypothesis that the α-cyclodextrin selectivity of Paenibacillus macerans α-CGTase could be augmented by masking subsite −7 of the active site, blocking the formation of larger cyclodextrins is tested.

Mutations of Lysine 47 in cyclodextrin glycosyltransferase from Paenibacillus macerans enhance beta-cyclodextrin specificity.

The effects of amino acid side chain at position 47 on cyclodextrin product specificity were investigated by replacing Lys47 in the CGTase from Paenibacillus macerans strain JFB05-01 with arginine, histidine, threonine, serine, or leucine, and all of the mutants displayed a shift in product specificity toward the production of beta-cyclodesxtrin.

Molecular modification of Bacillus stearothermophilus NO2 cyclodextrin glucosyltransferase and preparation of α-cyclodextrin

This research focuses on CGTase reaction and product specificities, which suggest a novel method for the industrial production of α-CD.

Rational design to change product specificities and thermostability of cyclodextrin glycosyltransferase from Paenibacillus sp.

Ala315 mutants were designed to improve thermostability, and A315D mutant exhibited the highest thermostable at 60–70 °C for 30 min, providing an effective approach for improving the product specificity and thermostabilities of CGTases.

Engineering cyclodextrin glycosyltransferase into a starch hydrolase with a high exo-specificity.

Y93F Substitution of Cyclodextrin Glucanotransferase from Bacillus sp. A2-5a and its Enzyme Characterization

Y93F substitution of BA2-5a CGTase alters the enzyme kinetic property and product specificity and the Y93F produced more β-CD and decreased its kcat and kcat/Km.

Double mutations enhance β-cyclization activity of cyclodextrin glycosyltransferase from Bacillus circulans

A CGTase with high coupling activity using γ-cyclodextrin isolated from a novel strain clustering under the genus Carboxydocella.

This enzyme is the first glycoside hydrolase isolated from the genus, indicating starch degradation via cyclodextrin production in the Carboxydocella strain, and its fundamental reactivities are characterized and compared with two commercial CGTases, assayed under identical condition, in order to facilitate interpretation of the results.

Enhancement of α-cyclodextrin product specificity by enriching histidines of α-cyclodextrin glucanotransferase at remote subsite −6

...

References

SHOWING 1-10 OF 55 REFERENCES

Site-directed mutations in tyrosine 195 of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 affect activity and product specificity.

The presence of aromatic amino acids (Tyr or Phe) at the Tyr195 position thus appears to be of crucial importance for an efficient cyclization reaction, virtually preventing the formation of linear products.

Engineering of factors determining alpha-amylase and cyclodextrin glycosyltransferase specificity in the cyclodextrin glycosyltransferase from Thermoanaerobacterium thermosulfurigenes EM1.

Characterization of the F196G mutant CGTase of T. thermosulfurigenes EM1 revealed that cyclization, coupling and disproportionation activities had become drastically reduced in the mutant F 196G(delta'DE), but its saccharifying activity had doubled, reaching the highest level ever reported for a CGT enzyme.

Engineering of Cyclodextrin Product Specificity and pH Optima of the Thermostable Cyclodextrin Glycosyltransferase fromThermoanaerobacterium thermosulfurigenes EM1*

The mutants showed changes in both the high and low pH slopes of the optimum curve for cyclization and hydrolysis when compared with the wild-type enzyme, which suggests that the pH optimum curve of CGTase is determined only by residue Glu258.

Four aromatic residues in the active center of cyclodextrin glucanotransferase from alkalophilic Bacillus sp. 1011: effects of replacements on substrate binding and cyclization characteristics.

Results indicated that Phe-183 and P he-259 are cooperatively involved in acceptor binding, and that they play a critical role in cyclization when the nonreducing end of amylose binds to the active center of CGTase.

Structure of cyclodextrin glycosyltransferase complexed with a maltononaose inhibitor at 2.6 angstrom resolution. Implications for product specificity.

The refined structure of the resulting protein-carbohydrate complex has final crystallographic and free R-factors for data in the 8-2.6 angstrom resolution range and reveals that a new inhibitor, composed of nine saccharide residues, is bound in the active site.

Replacement of an amino acid residue of cyclodextrin glucanotransferase of Bacillus ohbensis doubles the production of gamma-cyclodextrin.

Nucleotide sequence and X-ray structure of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 in a maltose-dependent crystal form.

The cyclodextrin glycosyltransferase (CGTase, EC 2.4.1.19) gene from Bacillus circulans strain 251 was cloned and sequenced. It was found to code for a mature protein of 686 amino acid residues,…

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

Two mutations were designed that changed the production spectrum from the predominant product beta-cyclodextrin of the wild-type enzyme towards gamma-cyclodesxtrin, which is of practical interest because it is rare and can encapsulate larger nonpolar compounds.

The three transglycosylation reactions catalyzed by cyclodextrin glycosyltransferase from Bacillus circulans (strain 251) proceed via different kinetic mechanisms.

The rate of interconversion of linear and circular conformations of oligosaccharides in the cyclization and coupling reactions was found to determine the reaction rate and will allow rational design of CGTase mutant enzymes synthesizing cyclodextrins of specific sizes.

The Raw Starch Binding Domain of Cyclodextrin Glycosyltransferase from Bacillus circulans Strain 251*

Purified mutant CGTases were characterized with respect to raw starch binding and cyclization reaction kinetics on both soluble and raw starch, and it is shown that maltose binding site 1 is most important for raw starchbinding, whereas maltosebinding site 2 is involved in guiding linear starch chains into the active site.
...