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Structure of two fungal beta-1,4-galactanases: searching for the basis for temperature and pH optimum.

Le Nours J, Ryttersgaard C, Lo Leggio L, Østergaard PR, Borchert TV, Christensen LL, Larsen S
Protein Sci 12 1195-204 (2003)
Related entries: 1hjq, 1hju

Cited: 27 times
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Abstract

beta-1,4-Galactanases hydrolyze the galactan side chains that are part of the complex carbohydrate structure of the pectin. They are assigned to family 53 of the glycoside hydrolases and display significant variations in their pH and temperature optimum and stability. Two fungal beta-1,4-galactanases from Myceliophthora thermophila and Humicola insolens have been cloned and heterologously expressed, and the crystal structures of the gene products were determined. The structures are compared to the previously only known family 53 structure of the galactanase from Aspergillus aculeatus (AAGAL) showing approximately 56% identity. The M. thermophila and H. insolens galactanases are thermophilic enzymes and are most active at neutral to basic pH, whereas AAGAL is mesophilic and most active at acidic pH. The structure of the M. thermophila galactanase (MTGAL) was determined from crystals obtained with HEPES and TRIS buffers to 1.88 A and 2.14 A resolution, respectively. The structure of the H. insolens galactanase (HIGAL) was determined to 2.55 A resolution. The thermostability of MTGAL and HIGAL correlates with increase in the protein rigidity and electrostatic interactions, stabilization of the alpha-helices, and a tighter packing. An inspection of the active sites in the three enzymes identifies several amino acid substitutions that could explain the variation in pH optimum. Examination of the activity as a function of pH for the D182N mutant of AAGAL and the A90S/ H91D mutant of MTGAL showed that the difference in pH optimum between AAGAL and MTGAL is at least partially associated with differences in the nature of residues at positions 182, 90, and/or 91.

Articles - 1hjs mentioned but not cited (5)

  1. Structure of two fungal beta-1,4-galactanases: searching for the basis for temperature and pH optimum. Le Nours J, Ryttersgaard C, Lo Leggio L, Østergaard PR, Borchert TV, Christensen LL, Larsen S. Protein Sci. 12 1195-1204 (2003)
  2. Structure of Aspergillus aculeatus β-1,4-galactanase in complex with galactobiose. Torpenholt S, Poulsen JCN, Muderspach SJ, De Maria L, Lo Leggio L. Acta Crystallogr F Struct Biol Commun 75 399-404 (2019)
  3. The binding of zinc ions to Emericella nidulans endo-β-1,4-galactanase is essential for crystal formation. Otten H, Michalak M, Mikkelsen JD, Larsen S. Acta Crystallogr. Sect. F Struct. Biol. Cryst. Commun. 69 850-854 (2013)
  4. Effect of mutations on the thermostability of Aspergillus aculeatus β-1,4-galactanase. Torpenholt S, De Maria L, Olsson MH, Christensen LH, Skjøt M, Westh P, Jensen JH, Lo Leggio L. Comput Struct Biotechnol J 13 256-264 (2015)
  5. Engineering the substrate binding site of the hyperthermostable archaeal endo-β-1,4-galactanase from Ignisphaera aggregans. Muderspach SJ, Fredslund F, Volf V, Poulsen JN, Blicher TH, Clausen MH, Rasmussen KK, Krogh KBRM, Jensen K, Lo Leggio L. Biotechnol Biofuels 14 183 (2021)


Reviews citing this publication (2)

  1. Myceliophthora thermophila syn. Sporotrichum thermophile: a thermophilic mould of biotechnological potential. Singh B. Crit. Rev. Biotechnol. 36 59-69 (2016)
  2. Peculiarities and applications of galactanolytic enzymes that act on type I and II arabinogalactans. Sakamoto T, Ishimaru M. Appl. Microbiol. Biotechnol. 97 5201-5213 (2013)

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  1. Comparative genome analysis of filamentous fungi reveals gene family expansions associated with fungal pathogenesis. Soanes DM, Alam I, Cornell M, Wong HM, Hedeler C, Paton NW, Rattray M, Hubbard SJ, Oliver SG, Talbot NJ. PLoS ONE 3 e2300 (2008)
  2. Redesigning protein pKa values. Tynan-Connolly BM, Nielsen JE. Protein Sci. 16 239-249 (2007)
  3. Bifidobacterium longum endogalactanase liberates galactotriose from type I galactans. Hinz SW, Pastink MI, van den Broek LA, Vincken JP, Voragen AG. Appl. Environ. Microbiol. 71 5501-5510 (2005)
  4. An alkaline active xylanase: insights into mechanisms of high pH catalytic adaptation. Mamo G, Thunnissen M, Hatti-Kaul R, Mattiasson B. Biochimie 91 1187-1196 (2009)
  5. Gene cloning, expression, and biochemical characterization of an alkali-tolerant β-mannanase from Humicola insolens Y1. Luo H, Wang K, Huang H, Shi P, Yang P, Yao B. J. Ind. Microbiol. Biotechnol. 39 547-555 (2012)
  6. Consistent mutational paths predict eukaryotic thermostability. van Noort V, Bradatsch B, Arumugam M, Amlacher S, Bange G, Creevey C, Falk S, Mende DR, Sinning I, Hurt E, Bork P. BMC Evol. Biol. 13 7 (2013)
  7. Purification and characterization of recombinant endoglucanase of Trichoderma reesei expressed in Saccharomyces cerevisiae with higher glycosylation and stability. Qin Y, Wei X, Liu X, Wang T, Qu Y. Protein Expr. Purif. 58 162-167 (2008)
  8. The structure of endo-beta-1,4-galactanase from Bacillus licheniformis in complex with two oligosaccharide products. Ryttersgaard C, Le Nours J, Lo Leggio L, Jørgensen CT, Christensen LL, Bjørnvad M, Larsen S. J. Mol. Biol. 341 107-117 (2004)
  9. Random exchanges of non-conserved amino acid residues among four parental termite cellulases by family shuffling improved thermostability. Ni J, Takehara M, Miyazawa M, Watanabe H. Protein Eng. Des. Sel. 20 535-542 (2007)
  10. Genetic and biochemical characterization of a protease-resistant mesophilic β-mannanase from Streptomyces sp. S27. Shi P, Yuan T, Zhao J, Huang H, Luo H, Meng K, Wang Y, Yao B. J. Ind. Microbiol. Biotechnol. 38 451-458 (2011)
  11. Characterization of a thermostable endo-beta-1,4-D-galactanase from the hyperthermophile Thermotoga maritima. Yang H, Ichinose H, Yoshida M, Nakajima M, Kobayashi H, Kaneko S. Biosci. Biotechnol. Biochem. 70 538-541 (2006)
  12. Structural and biochemical studies elucidate the mechanism of rhamnogalacturonan lyase from Aspergillus aculeatus. Jensen MH, Otten H, Christensen U, Borchert TV, Christensen LL, Larsen S, Leggio LL. J. Mol. Biol. 404 100-111 (2010)
  13. Activity of three β-1,4-galactanases on small chromogenic substrates. Torpenholt S, Le Nours J, Christensen U, Jahn M, Withers S, Ostergaard PR, Borchert TV, Poulsen JC, Lo Leggio L. Carbohydr. Res. 346 2028-2033 (2011)
  14. Alkalophilic adaptation of XynB endoxylanase from Aspergillus niger via rational design of pKa of catalytic residues. Xu H, Zhang F, Shang H, Li X, Wang J, Qiao D, Cao Y. J. Biosci. Bioeng. 115 618-622 (2013)
  15. Investigating the binding of beta-1,4-galactan to Bacillus licheniformis beta-1,4-galactanase by crystallography and computational modeling. Le Nours J, De Maria L, Welner D, Jørgensen CT, Christensen LL, Borchert TV, Larsen S, Lo Leggio L. Proteins 75 977-989 (2009)
  16. Thermostability enhancement of an endo-1,4-β-galactanase from Talaromyces stipitatus by site-directed mutagenesis. Larsen DM, Nyffenegger C, Swiniarska MM, Thygesen A, Strube ML, Meyer AS, Mikkelsen JD. Appl. Microbiol. Biotechnol. 99 4245-4253 (2015)
  17. Improving the acidic stability of Staphylococcus aureus α-acetolactate decarboxylase in Bacillus subtilis by changing basic residues to acidic residues. Zhang X, Rao Z, Li J, Zhou J, Yang T, Xu M, Bao T, Zhao X. Amino Acids 47 707-717 (2015)
  18. Structure-Based Design of Acetolactate Synthase From Bacillus licheniformis Improved Protein Stability Under Acidic Conditions. Zhao T, Li Y, Yuan S, Ye Y, Peng Z, Zhou R, Liu J. Front Microbiol 11 582909 (2020)
  19. Penicillium purpurogenum produces a highly stable endo-β-(1,4)-galactanase. Zavaleta V, Eyzaguirre J. Appl. Biochem. Biotechnol. 180 1313-1327 (2016)
  20. The use of Agrobacterium-mediated insertional mutagenesis sequencing to identify novel genes of Humicola insolens involved in cellulase production. Fan C, Xu X, Song L, Guan W, Li J, Liu B, Shi P, Zhang W. 3 Biotech 8 153 (2018)


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