1cgv › Citations

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

Penninga D, Strokopytov B, Rozeboom HJ, Lawson CL, Dijkstra BW, Bergsma J, Dijkhuizen L
Biochemistry 34 3368-76 (1995)
Related entries: 1cgw, 1cgx, 1cgy

Cited: 62 times
EuropePMC logo PMID: 7880832

Abstract

Tyrosine 195 is located in the center of the active site cleft of cyclodextrin glycosyltransferase (EC 2.4.1.19) from Bacillus circulans strain 251. Alignment of amino acid sequences of CGTases and alpha-amylases, and the analysis of the binding mode of the substrate analogue acarbose in the active site cleft [Strokopytov, B., et al. (1995) Biochemistry 34, (in press)], suggested that Tyr195 plays an important role in cyclization of oligosaccharides. Tyr195 therefore was replaced with Phe (Y195F), Trp (Y195W), Leu (Y195L), and Gly (Y195G). Mutant proteins were purified and crystallized, and their X-ray structures were determined at 2.5-2.6 angstrum resolution, allowing a detailed comparison of their biochemical properties and three-dimensional structures with those of the wild-type CGTase protein. The mutant proteins possessed significantly reduced cyclodextrin forming and coupling activities but were not negatively affected in the disproportionation and saccharifying reactions. Also under production process conditions, after a 45 h incubation with a 10% starch solution, the Y195W, Y195L, and Y195G mutants showed a lower overall conversion of starch into cyclodextrins. These mutants produced a considerable amount of linear maltooligosaccharides. 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. Mass spectrometry of the Y195L reaction mixture, but not that of the other mutants and the wild type, revealed a shift toward the synthesis (in low yields) of larger products, especially of beta- and gamma- (but no alpha-) cyclodextrins and minor amounts of delta-, epsilon-, zeta- and eta-cyclodextrins.(ABSTRACT TRUNCATED AT 250 WORDS)

Articles - 1cgv mentioned but not cited (1)

  1. Local functional descriptors for surface comparison based binding prediction. Cipriano GM, Phillips GN, Gleicher M. BMC Bioinformatics 13 314 (2012)


Reviews citing this publication (11)

  1. The carbohydrate-binding module family 20--diversity, structure, and function. Christiansen C, Abou Hachem M, Janecek S, Viksø-Nielsen A, Blennow A, Svensson B. FEBS J 276 5006-5029 (2009)
  2. Protein engineering of bacterial alpha-amylases. Nielsen JE, Borchert TV. Biochim Biophys Acta 1543 253-274 (2000)
  3. alpha-Amylase family: molecular biology and evolution. Janecek S. Prog Biophys Mol Biol 67 67-97 (1997)
  4. Thiooligosaccharides as tools for structural biology. Driguez H. Chembiochem 2 311-318 (2001)
  5. gamma-Cyclodextrin: a review on enzymatic production and applications. Li Z, Wang M, Wang F, Gu Z, Du G, Wu J, Chen J. Appl Microbiol Biotechnol 77 245-255 (2007)
  6. Engineering of cyclodextrin glucanotransferases and the impact for biotechnological applications. Leemhuis H, Kelly RM, Dijkhuizen L. Appl Microbiol Biotechnol 85 823-835 (2010)
  7. Cyclodextrin glucanotransferase: from gene to applications. Qi Q, Zimmermann W. Appl Microbiol Biotechnol 66 475-485 (2005)
  8. The functions of 4-alpha-glucanotransferases and their use for the production of cyclic glucans. Takaha T, Smith SM. Biotechnol Genet Eng Rev 16 257-280 (1999)
  9. Recent advances in discovery, heterologous expression, and molecular engineering of cyclodextrin glycosyltransferase for versatile applications. Han R, Li J, Shin HD, Chen RR, Du G, Liu L, Chen J. Biotechnol Adv 32 415-428 (2014)
  10. Comprehensive study on transglycosylation of CGTase from various sources. Lim CH, Rasti B, Sulistyo J, Hamid MA. Heliyon 7 e06305 (2021)
  11. Strategies for enhancing extracellular secretion of recombinant cyclodextrin glucanotransferase in E. coli. Tesfai BT, Wu D, Chen S, Chen J, Wu J. Appl Biochem Biotechnol 167 897-908 (2012)

Articles citing this publication (50)

  1. The three transglycosylation reactions catalyzed by cyclodextrin glycosyltransferase from Bacillus circulans (strain 251) proceed via different kinetic mechanisms. van der Veen BA, van Alebeek GJ, Uitdehaag JC, Dijkstra BW, Dijkhuizen L. Eur J Biochem 267 658-665 (2000)
  2. Crystal structure of amylomaltase from thermus aquaticus, a glycosyltransferase catalysing the production of large cyclic glucans. Przylas I, Tomoo K, Terada Y, Takaha T, Fujii K, Saenger W, Sträter N. J Mol Biol 296 873-886 (2000)
  3. Rational design of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 to increase alpha-cyclodextrin production. van der Veen BA, Uitdehaag JC, Penninga D, van Alebeek GJ, Smith LM, Dijkstra BW, Dijkhuizen L. J Mol Biol 296 1027-1038 (2000)
  4. The evolution of cyclodextrin glucanotransferase product specificity. Kelly RM, Dijkhuizen L, Leemhuis H. Appl Microbiol Biotechnol 84 119-133 (2009)
  5. Mutations converting cyclodextrin glycosyltransferase from a transglycosylase into a starch hydrolase. Leemhuis H, Dijkstra BW, Dijkhuizen L. FEBS Lett 514 189-192 (2002)
  6. Crystal structure of a compact α-amylase from Geobacillus thermoleovorans. Mok SC, Teh AH, Saito JA, Najimudin N, Alam M. Enzyme Microb Technol 53 46-54 (2013)
  7. The role of arginine 47 in the cyclization and coupling reactions of cyclodextrin glycosyltransferase from Bacillus circulans strain 251 implications for product inhibition and product specificity. van der Veen BA, Uitdehaag JC, Dijkstra BW, Dijkhuizen L. Eur J Biochem 267 3432-3441 (2000)
  8. Identification, cloning, expression, and characterization of the extracellular acarbose-modifying glycosyltransferase, AcbD, from Actinoplanes sp. strain SE50. Hemker M, Stratmann A, Goeke K, Schröder W, Lenz J, Piepersberg W, Pape H. J Bacteriol 183 4484-4492 (2001)
  9. Addition of polar organic solvents can improve the product selectivity of cyclodextrin glycosyltransferase. Solvent effects on cgtase. Blackwood AD, Bucke C. Enzyme Microb Technol 27 704-708 (2000)
  10. Mutations at subsite -3 in cyclodextrin glycosyltransferase from Paenibacillus macerans enhancing alpha-cyclodextrin specificity. Li Z, Zhang J, Wang M, Gu Z, Du G, Li J, Wu J, Chen J. Appl Microbiol Biotechnol 83 483-490 (2009)
  11. Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp. G1 in E. coli. Ong RM, Goh KM, Mahadi NM, Hassan O, Rahman RN, Illias RM. J Ind Microbiol Biotechnol 35 1705-1714 (2008)
  12. Conversion of the maltogenic alpha-amylase Novamyl into a CGTase. Beier L, Svendsen A, Andersen C, Frandsen TP, Borchert TV, Cherry JR. Protein Eng 13 509-513 (2000)
  13. Improved thermostability of bacillus circulans cyclodextrin glycosyltransferase by the introduction of a salt bridge. Leemhuis H, Rozeboom HJ, Dijkstra BW, Dijkhuizen L. Proteins 54 128-134 (2004)
  14. Cyclodextrin glycosyltransferase: a key enzyme in the assimilation of starch by the halophilic archaeon Haloferax mediterranei. Bautista V, Esclapez J, Pérez-Pomares F, Martínez-Espinosa RM, Camacho M, Bonete MJ. Extremophiles 16 147-159 (2012)
  15. Use of random and saturation mutageneses to improve the properties of Thermus aquaticus amylomaltase for efficient production of cycloamyloses. Fujii K, Minagawa H, Terada Y, Takaha T, Kuriki T, Shimada J, Kaneko H. Appl Environ Microbiol 71 5823-5827 (2005)
  16. Purification and properties of a novel raw starch degrading-cyclodextrin glycosyltransferase from Klebsiella pneumoniae AS- 22. Gawande BN, Patkar AY. Enzyme Microb Technol 28 735-743 (2001)
  17. 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. Han R, Liu L, Shin HD, Chen RR, Li J, Du G, Chen J. Appl Environ Microbiol 79 672-677 (2013)
  18. Enzymatic circularization of a malto-octaose linear chain studied by stochastic reaction path calculations on cyclodextrin glycosyltransferase. Uitdehaag JC, van der Veen BA, Dijkhuizen L, Elber R, Dijkstra BW. Proteins 43 327-335 (2001)
  19. Structural elucidation of the cyclization mechanism of α-1,6-glucan by Bacillus circulans T-3040 cycloisomaltooligosaccharide glucanotransferase. Suzuki N, Fujimoto Z, Kim YM, Momma M, Kishine N, Suzuki R, Suzuki S, Kitamura S, Kobayashi M, Kimura A, Funane K. J Biol Chem 289 12040-12051 (2014)
  20. Comparative study of the cyclization reactions of three bacterial cyclomaltodextrin glucanotransferases. Terada Y, Sanbe H, Takaha T, Kitahata S, Koizumi K, Okada S. Appl Environ Microbiol 67 1453-1460 (2001)
  21. Construction of chimeric cyclodextrin glucanotransferases from Bacillus circulans A11 and Paenibacillus macerans IAM1243 and analysis of their product specificity. Rimphanitchayakit V, Tonozuka T, Sakano Y. Carbohydr Res 340 2279-2289 (2005)
  22. Site-saturation engineering of lysine 47 in cyclodextrin glycosyltransferase from Paenibacillus macerans to enhance substrate specificity towards maltodextrin for enzymatic synthesis of 2-O-D-glucopyranosyl-L-ascorbic acid (AA-2G). Han R, Liu L, Shin HD, Chen RR, Du G, Chen J. Appl Microbiol Biotechnol 97 5851-5860 (2013)
  23. Chemical modification of lysine side chains of cyclodextrin glycosyltransferase from Thermoanaerobacter causes a shift from cyclodextrin glycosyltransferase to alpha-amylase specificity. Alcalde M, Plou FJ, Andersen C, Martín MT, Pedersen S, Ballesteros A. FEBS Lett 445 333-337 (1999)
  24. Carbohydrate-binding module-cyclodextrin glycosyltransferase fusion enables efficient synthesis of 2-O-d-glucopyranosyl-l-ascorbic acid with soluble starch as the glycosyl donor. Han R, Li J, Shin HD, Chen RR, Du G, Liu L, Chen J. Appl Environ Microbiol 79 3234-3240 (2013)
  25. The fully conserved Asp residue in conserved sequence region I of the alpha-amylase family is crucial for the catalytic site architecture and activity. Leemhuis H, Rozeboom HJ, Dijkstra BW, Dijkhuizen L. FEBS Lett 541 47-51 (2003)
  26. Thermoanaerobacterium thermosulfurigenes cyclodextrin glycosyltransferase. Leemhuis H, Dijkstra BW, Dijkhuizen L. Eur J Biochem 270 155-162 (2003)
  27. Site-saturation mutagenesis of central tyrosine 195 leading to diverse product specificities of an α-cyclodextrin glycosyltransferase from Paenibacillus sp. 602-1. Xie T, Song B, Yue Y, Chao Y, Qian S. J Biotechnol 170 10-16 (2014)
  28. Truncated human serum albumin retains general anaesthetic binding activity. Liu R, Yang J, Ha CE, Bhagavan NV, Eckenhoff RG. Biochem J 388 39-45 (2005)
  29. Engineered cyclodextrin glucanotransferases from Bacillus sp. G-825-6 produce large-ring cyclodextrins with high specificity. Sonnendecker C, Melzer S, Zimmermann W. Microbiologyopen 8 e00757 (2019)
  30. Improving maltodextrin specificity for enzymatic synthesis of 2-O-d-glucopyranosyl-l-ascorbic acid by site-saturation engineering of subsite-3 in cyclodextrin glycosyltransferase from Paenibacillus macerans. Liu L, Xu Q, Han R, Shin HD, Chen RR, Li J, Du G, Chen J. J Biotechnol 166 198-205 (2013)
  31. A CGTase with high coupling activity using γ-cyclodextrin isolated from a novel strain clustering under the genus Carboxydocella. Ara KZ, Lundemo P, Fridjonsson OH, Hreggvidsson GO, Adlercreutz P, Karlsson EN. Glycobiology 25 514-523 (2015)
  32. Alteration of product specificity of cyclodextrin glucanotransferase from Thermococcus sp. B1001 by site-directed mutagenesis. Yamamoto T, Fujiwara S, Tachibana Y, Takagi M, Fukui K, Imanaka T. J Biosci Bioeng 89 206-209 (2000)
  33. Improved activity of β-cyclodextrin glycosyltransferase from Bacillus sp. N-227 via mutagenesis of the conserved residues. Wang H, Zhou W, Li H, Rie B, Piao C. 3 Biotech 7 149 (2017)
  34. Polyethylene glycols enhance the thermostability of β-cyclodextrin glycosyltransferase from Bacillus circulans. Li C, Li W, Holler TP, Gu Z, Li Z. Food Chem 164 17-22 (2014)
  35. Structural and Functional Characterization of Three Novel Fungal Amylases with Enhanced Stability and pH Tolerance. Roth C, Moroz OV, Turkenburg JP, Blagova E, Waterman J, Ariza A, Ming L, Tianqi S, Andersen C, Davies GJ, Wilson KS. Int J Mol Sci 20 E4902 (2019)
  36. An active-site mutation causes enhanced reactivity and altered regiospecificity of transglucosylation catalyzed by the Bacillus sp. SAM1606 alpha-glucosidase. Inohara-Ochiai M, Okada M, Nakayama T, Hemmi H, Ueda T, Iwashita T, Kan Y, Shibano Y, Ashikari T, Nishino T. J Biosci Bioeng 89 431-437 (2000)
  37. Bacillus macerans cyclomaltodextrin glucanotransferase transglycosylation reactions with different molar ratios of D-glucose and cyclomaltohexaose. Yoon SH, Robyt JF. Carbohydr Res 337 2245-2254 (2002)
  38. Succinylation of cyclodextrin glycosyltransferase from Thermoanaerobacter sp. 501 enhances its transferase activity using starch as donor. Alcalde M, Plou FJ, Teresa Martín M, Valdés I, Méndez E, Ballesteros A. J Biotechnol 86 71-80 (2001)
  39. Biosynthesis of 2-O-D-glucopyranosyl-l-ascorbic acid from maltose by an engineered cyclodextrin glycosyltransferase from Paenibacillus macerans. Liu L, Han R, Shin HD, Li J, Du G, Chen J. Carbohydr Res 382 101-107 (2013)
  40. CGTase-catalysed cis-glucosylation of L-rhamnosides for the preparation of Shigella flexneri 2a and 3a haptens. Urbach C, Halila S, Guerreiro C, Driguez H, Mulard LA, Armand S. Chembiochem 15 293-300 (2014)
  41. Engineering CGTase to improve synthesis of alkyl glycosides. Ara KZG, Linares-Pastén JA, Jönsson J, Viloria-Cols M, Ulvenlund S, Adlercreutz P, Karlsson EN. Glycobiology 31 603-612 (2021)
  42. Engineering of Cyclodextrin Glycosyltransferase through a Size/Polarity Guided Triple-Code Strategy with Enhanced α-Glycosyl Hesperidin Synthesis Ability. Chen H, Liu Y, Ren X, Wang J, Zhu L, Lu Y, Chen X. Appl Environ Microbiol 88 e0102722 (2022)
  43. Transfer reactions catalyzed by cyclodextrin glucosyltransferase using 4-thiomaltosyl and C-maltosyl fluorides as artificial donors. Bornaghi L, Utille JP, Rekaï el-D, Mallet JM, Sinaÿ P, Driguez H. Carbohydr Res 305 561-568 (1997)
  44. Molecular dynamic analysis of mutant Y195I α-cyclodextrin glycosyltransferase with switched product specificity from α-cyclodextrin to γ-cyclodextrin. Chen F, Xie T, Yue Y, Qian S, Chao Y, Pei J. J Mol Model 21 208 (2015)
  45. Characterisation of a glycosylated alkyl polyglycoside produced by a cyclodextrin glycosyltransferase by HPLC-ELSD and -MS. Svensson D, Adlercreutz P. J Chromatogr B Analyt Technol Biomed Life Sci 879 1857-1860 (2011)
  46. Effect of chemical modification of cyclodextrin glycosyltransferase (CGTase) from Thermoanaerobacter sp. on its activity and product selectivity. Alcalde M, Plou FJ, Pastor E, Ballesteros A. Ann N Y Acad Sci 864 183-187 (1998)
  47. Engineering of cyclodextrin glycosyltransferase improves the conversion efficiency of rebaudioside A to glucosylated steviol glycosides and increases the content of short-chain glycosylated steviol glycoside. Zhang R, Tang R, Wang W, Bi J, Xu X, Fan Q, Li Y, Chen Q. Microb Cell Fact 22 113 (2023)
  48. Structural basis of a mutant Y195I α-cyclodextrin glycosyltransferase with switched product specificity from α-cyclodextrin to β-/γ-cyclodextrin. Xie T, Hou Y, Li D, Yue Y, Qian S, Chao Y. J Biotechnol 182-183 92-96 (2014)
  49. Enhancing 2-O-α-D-glucopyranosyl-L-ascorbic acid synthesis by weakening the acceptor specificity of CGTase toward glucose and maltose. Tao X, Kong D, Zhang H, Su L, Chen S, Rao D, Wei B, Wu J, Wang L. Bioprocess Biosyst Eng 46 903-911 (2023)
  50. Optimized enzymatic synthesis of digestive resistant anomalous isoquercitrin glucosides using amylosucrase and response surface methodology. Rha CS, Park CS, Kim DO. Appl Microbiol Biotechnol 105 6931-6941 (2021)


Related citations provided by authors (2)

  1. Nucleotide Sequence and X-Ray Structure of Cyclodextrin Glycosyltransferase from Bacillus Circulans Strain 251 in a Maltose-Dependent Crystal Form. Lawson CLL, Van Montfort R, Strokopytov B, Rozeboom HJ, Kalk KH, De Vries GE, Penninga D, Dijkhuizen L, Dijkstra BW J. Mol. Biol. 236 590- (1994)
  2. Maltodextrin-Dependent Crystallization of Cyclomaltodextrin Glucanotransferase from Bacillus Circulans. Lawson CLL, Bergsma J, Bruinenberg PM, De Vries GE, Dijkhuizen L, Dijkstra BW J. Mol. Biol. 214 807- (1990)

Quick links