Kinetics of substrate transglycosylation by glycoside hydrolase family 3 glucan (1→3)-β-glucosidase from the white-rot fungus Phanerochaete chrysosporium

2004 ◽  
Vol 339 (18) ◽  
pp. 2851-2857 ◽  
Author(s):  
Rie Kawai ◽  
Kiyohiko Igarashi ◽  
Motomitsu Kitaoka ◽  
Tadashi Ishii ◽  
Masahiro Samejima
Keyword(s):  
Phanerochaete Chrysosporium ◽  
Glycoside Hydrolase ◽  
White Rot ◽  
White Rot Fungus ◽  
Glycoside Hydrolase Family ◽  
Kinetics Of ◽  
Hydrolase Family

scholarly journals Transcript Analysis of Genes Encoding a Family 61 Endoglucanase and a Putative Membrane-Anchored Family 9 Glycosyl Hydrolase from Phanerochaete chrysosporium

2002 ◽  
Vol 68 (11) ◽  
pp. 5765-5768 ◽  
Author(s):  
Amber Vanden Wymelenberg ◽  
Stuart Denman ◽  
Diane Dietrich ◽  
Jennifer Bassett ◽  
Xiaochun Yu ◽  
...  
Keyword(s):  
Phanerochaete Chrysosporium ◽  
Glycoside Hydrolase ◽  
Glycosyl Hydrolase ◽  
Thermobifida Fusca ◽  
Glycoside Hydrolase Family ◽  
Transcript Analysis ◽  
Transcript Levels ◽  
Genes Encoding ◽  
Membrane Bound ◽  
Hydrolase Family

ABSTRACT Phanerochaete chrysosporium cellulase genes were cloned and characterized. The cel61A product was structurally similar to fungal endoglucanases of glycoside hydrolase family 61, whereas the cel9A product revealed similarities to Thermobifida fusca Cel9A (E4), an enzyme with both endo- and exocellulase characteristics. The fungal Cel9A is apparently a membrane-bound protein, which is very unusual for microbial cellulases. Transcript levels of both genes were substantially higher in cellulose-grown cultures than in glucose-grown cultures. These results show that P. chrysosporium possesses a wide array of conventional and unconventional cellulase genes.

scholarly journals Thermostable Mutants of Glycoside Hydrolase Family 6 Cellobiohydrolase from the Basidiomycete Phanerochaete chrysosporium

2020 ◽  
Vol 67 (3) ◽  
pp. 79-86
Author(s):  
Sora Yamaguchi ◽  
Naoki Sunagawa ◽  
Mikako Tachioka ◽  
Kiyohiko Igarashi ◽  
Masahiro Samejima
Keyword(s):  
Phanerochaete Chrysosporium ◽  
Glycoside Hydrolase ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family

scholarly journals Thermostable mutants of glycoside hydrolase family 6 cellobiohydrolase from the basidiomycete Phanerochaete chrysosporium

2020 ◽  
Author(s):  
Sora Yamaguchi ◽  
Naoki Sunagawa ◽  
Mikako Tachioka ◽  
Kiyohiko Igarashi ◽  
Masahiro Samejima
Keyword(s):  
Thermal Stability ◽  
Phanerochaete Chrysosporium ◽  
Glycoside Hydrolase ◽  
Thermal Inactivation ◽  
Renewable Resource ◽  
Cellulosic Biomass ◽  
Limiting Factor ◽  
Glycoside Hydrolase Family ◽  
Hydrolytic Activities ◽  
Hydrolase Family

AbstractThermal inactivation of saccharifying enzymes is a crucial issue for the efficient utilization of cellulosic biomass as a renewable resource. Cellobiohydrolases (CBHs) is a kind of cellulase. In general, CBHs belonging to glycoside hydrolase (GH) family 6 (Cel6) act synergistically with CBHs of GH family 7 (Cel7) and other carbohydrate-active enzymes during the degradation of cellulosic biomass. However, while the catalytic rate of enzymes generally becomes faster at higher temperatures, Cel6 CBHs are inactivated at lower temperatures than Cel7 CBHs, and this represents a limiting factor for industrial utilization. In this study, we produced a series of mutants of the glycoside hydrolase family 6 cellobiohydrolase PcCel6A from the fungus Phanerochaete chrysosporium, and compared their thermal stability. Eight mutants from a random mutagenesis library and one rationally designed mutant were selected as candidate thermostable mutants and produced by heterologous expression in the yeast Pichia pastoris. Comparison of the hydrolytic activities at 50 and 60 °C indicated that the thermal stability of PcCel6A is influenced by the number and position of cysteine residues that are not involved in disulfide bonds.

scholarly journals Kinetics of inter-domain electron transfer in flavocytochrome cellobiose dehydrogenase from the white-rot fungus Phanerochaete chrysosporium

2002 ◽  
Vol 365 (2) ◽  
pp. 521-526 ◽  
Author(s):  
Kiyohiko IGARASHI ◽  
Ikuo MOMOHARA ◽  
Takeshi NISHINO ◽  
Masahiro SAMEJIMA
Keyword(s):  
Electron Transfer ◽  
Phanerochaete Chrysosporium ◽  
Substrate Concentration ◽  
Substrate Inhibition ◽  
Cellobiose Dehydrogenase ◽  
White Rot ◽  
White Rot Fungus ◽  
Isosbestic Point ◽  
Second Phase ◽  
Kinetics Of

The pre-steady-state kinetics of inter-domain electron transfer in the extracellular flavocytochrome cellobiose dehydrogenase from Phanerochaete chrysosporium was studied using various values of pH and substrate concentration. Monitoring at the isosbestic point of each prosthetic group indicated that the reductive half-reactions of flavin and haem were biphasic and monophasic respectively. When the observed rates of the flavin and haem reactions were plotted against substrate concentration, the behaviour of the second phase of the flavin reduction was almost identical with that of haem reduction at all substrate concentrations and pH values tested, suggesting that the formation of flavin semiquinone and haem reduction involve the same electron transfer reaction. Although flavin reduction by cellobiose was observed in the range of pH3.0–7.0, the velocity of the next electron transfer step decreased with increase of pH and was almost zero above pH6.0. The second phase of flavin reduction and the haem reduction were inhibited similarly by high concentrations of the substrate, whereas the first phase of flavin reduction showed a hyperbolic relation to the cellobiose concentration. Increase in pH enhanced the substrate inhibition of haem reduction but not the initial flavin reduction. Moreover, the dissociation constant Kd of flavin reduction and the substrate inhibition constant Ki of haem reduction decreased similarly with an increase of pH. From these results, it is evident that binding of cellobiose to the active site inhibits electron transfer from flavin to haem.

Atypical kinetics of cytochromes P450 catalysing 3'-hydroxylation of flavone from the white-rot fungus Phanerochaete chrysosporium

2009 ◽  
Vol 147 (1) ◽  
pp. 117-125 ◽  
Author(s):  
N. Kasai ◽  
S. Ikushiro ◽  
S. Hirosue ◽  
A. Arisawa ◽  
H. Ichinose ◽  
...  
Keyword(s):  
Phanerochaete Chrysosporium ◽  
Cytochromes P450 ◽  
White Rot ◽  
White Rot Fungus ◽  
Kinetics Of

scholarly journals Structure and function of Bs164 β-mannosidase from Bacteroides salyersiae the founding member of glycoside hydrolase family GH164

2019 ◽  
Vol 295 (13) ◽  
pp. 4316-4326 ◽  
Author(s):  
Zachary Armstrong ◽  
Gideon J. Davies
Keyword(s):  
Glycoside Hydrolase ◽  
Biochemical Characterization ◽  
Glycoside Hydrolase Family ◽  
Human Gut ◽  
X Ray Crystallography ◽  
Founding Member ◽  
And Function ◽  
Protein Sequence Space ◽  
Kinetics Of ◽  
Hydrolase Family

Recent work exploring protein sequence space has revealed a new glycoside hydrolase (GH) family (GH164) of putative mannosidases. GH164 genes are present in several commensal bacteria, implicating these genes in the degradation of dietary glycans. However, little is known about the structure, mechanism of action, and substrate specificity of these enzymes. Herein we report the biochemical characterization and crystal structures of the founding member of this family (Bs164) from the human gut symbiont Bacteroides salyersiae. Previous reports of this enzyme indicated that it has α-mannosidase activity, however, we conclusively show that it cleaves only β-mannose linkages. Using NMR spectroscopy, detailed enzyme kinetics of WT and mutant Bs164, and multiangle light scattering we found that it is a trimeric retaining β-mannosidase, that is susceptible to several known mannosidase inhibitors. X-ray crystallography revealed the structure of Bs164, the first known structure of a GH164, at 1.91 Å resolution. Bs164 is composed of three domains: a (β/α)8 barrel, a trimerization domain, and a β-sandwich domain, representing a previously unobserved structural-fold for β-mannosidases. Structures of Bs164 at 1.80–2.55 Å resolution in complex with the inhibitors noeuromycin, mannoimidazole, or 2,4-dinitrophenol 2-deoxy-2-fluoro-mannoside reveal the residues essential for specificity and catalysis including the catalytic nucleophile (Glu-297) and acid/base residue (Glu-160). These findings further our knowledge of the mechanisms commensal microbes use for nutrient acquisition.

scholarly journals Crystal structure of a family 6 cellobiohydrolase from the basidiomycetePhanerochaete chrysosporium

2017 ◽  
Vol 73 (7) ◽  
pp. 398-403 ◽  
Author(s):  
Mikako Tachioka ◽  
Akihiko Nakamura ◽  
Takuya Ishida ◽  
Kiyohiko Igarashi ◽  
Masahiro Samejima
Keyword(s):  
Catalytic Domain ◽  
White Rot ◽  
White Rot Fungus ◽  
Crystalline Cellulose ◽  
Glycoside Hydrolase Family ◽  
Carbohydrate Binding ◽  
Linker Peptide ◽  
Ligand Free ◽  
Hydrolysis Of ◽  
Hydrolase Family

Cellobiohydrolases belonging to glycoside hydrolase family 6 (CBH II, Cel6A) play key roles in the hydrolysis of crystalline cellulose. CBH II from the white-rot fungusPhanerochaete chrysosporium(PcCel6A) consists of a catalytic domain (CD) and a carbohydrate-binding module connected by a linker peptide, like other known fungal cellobiohydrolases. In the present study, the CD ofPcCel6A was crystallized without ligands, andp-nitrophenyl β-D-cellotrioside (pNPG3) was soaked into the crystals. The determined structures of the ligand-free andpNPG3-soaked crystals revealed that binding of cellobiose at substrate subsites +1 and +2 induces a conformational change of the N-terminal and C-terminal loops, switching the tunnel-shaped active site from the open to the closed form.

Sign in / Sign up

Export Citation Format

Share Document