CAZypedia - User contributions [en-ca]

Glycoside Hydrolase Family 65

2011-05-29T14:39:31Z

Hiroyuki Nakai:

{{CuratorApproved}}
* [[Author]]: ^^^Hiroyuki Nakai^^^
* [[Responsible Curator]]: ^^^Hiroyuki Nakai^^^
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<div style="float:right">
{| {{Prettytable}}
|-
|{{Hl2}} colspan="2" align="center" |'''Glycoside Hydrolase Family GH65'''
|-
|'''Clan'''
|GH-L
|-
|'''Mechanism'''
|inverting
|-
|'''Active site residues'''
|known
|-
|{{Hl2}} colspan="2" align="center" |'''CAZy DB link'''
|-
| colspan="2" |{{CAZyDBlink}}GH65.html
|}
</div>


== Substrate specificities ==
Glycoside hydrolase belonging to [[GH65]] acts on the substrate containing α-glucosidic linkages. [[GH65]] is mainly constructed by phosphorylase; maltose (Glc-α-1,4-Glc) phosphorylase (EC [{{EClink}}2.4.1.8 2.4.1.8]), trehalose (Glc-α1,α1-Glc) phosphorylase (EC [{{EClink}}2.4.1.64 2.4.1.64]), kojibiose (Glc-α-1,2-Glc) phosphorylase (EC [{{EClink}}2.4.1.230 2.4.1.230]), and trehalose 6-phosphate (Glc-α1,α1-Glc6P) phosphorylase (EC [{{EClink}}2.4.1.- 2.4.1.-]). Noticeably α,α-trehalase (EC [{{EClink}}3.2.1.28 3.2.1.28]) is also categorized from hydrolase as [[GH65]] members.

== Kinetics and Mechanism ==
Phosphorolysis by [[GH65]] enzymes proceeds with inversion of anomeric configuration, as first shown by Charlotte Fitting and Michael Doudoroff <cite>Fitting1952</cite> on maltose phosphorylase from ''Neisseria meningitidis'', i.e. maltose + Pi ↔ β-glucose 1-phosphate + glucose. The reaction mechanism for inverting [[GH65]] phosphorylase has been proposed to be similar to an genenal acid/base-catalysed direct displacement mechanism for inverting glycoside hydrolase <cite>Nakai2009</cite>, as proposed for inverting [[GH94]] phosphorylase (previously classified into glycoside transferase family 36) <cite>Hidaka2004</cite>, where direct nucleophilic attack by phosphate on the anomeric C-1 carbon is assisted by proton donation from the general acid catalyst to the glycosidic oxygen, instead of water molecule activated by general base catalyst in the inverting glycoside hydrolase reaction. The inverting phosphorolysis catalyzed by GH65 enzyme is reversible, which confers the phosphorylase with a capacity to effectively synthesize various α-glucosides from β-glucose 1-phosphate as donor and acceptor molecules. Noticeably β-glucosy fluoride can be used as donor in the synthetic reaction instead of the β-glucose 1-phosphate <cite>Tsumuraya1990</cite>.

== Catalytic Residues ==
The general acid catalyst was firstly predicted by superimposing the active site structure of maltose phosphorylase from ''Lactobacillus brevis'' <cite>Egloff2001</cite> with a catalytic (α/α)<sub>6</sub> barrel domain of GH15 glucoamylase (EC [{{EClink}}3.2.1.28 3.2.1.28]) from ''Aspergillus awamori'' <cite>Aleshin1992</cite>. Considering the similarities of the active site structure, Glu487 of ''L. brevis'' maltose phosphorylase was estimated as the general acid residue. Additionally it had been proved by site-direct mutagenesis on Glu487 of ''Paenibacillus'' sp. maltose phosphorylase <cite>Hidaka2005</cite>, which corresponds to the Glu487 of ''L. brevis'' maltose phosphorylase.

== Three-dimensional structures ==
The three-dimensional structure of ''L. brevis'' maltose phosphorylase (PDB ID [{{PDBlink}}1h54 1h54]) was determined <cite>Egloff2001</cite> and shows similarities with the (α/α)<sub>6</sub> barrel fold of [[GH15]] glucoamylase (EC [{{EClink}}3.2.1.28 3.2.1.28]) <cite>Aleshin1992</cite>, [[GH94]] cellobiose phosphorylase (EC [{{EClink}}2.4.1.20 2.4.1.20]) <cite>Hidaka2006</cite>, and [[GH94]] chitobiose phosphorylase (EC [{{EClink}}2.4.1.- 2.4.1.-]) <cite>Hidaka2004</cite>. [[GH15]] and [[GH65]] together constitute glycoside hydrolase clan L <cite>Cantarel2009</cite>.

== Family Firsts ==
;First stereochemistry determination: maltose phosphorylase (EC [{{EClink}}2.4.1.8 2.4.1.8]) from ''Neisseria meningitidis'' <cite>Fitting1952</cite>.
;First sequence identification: maltose phosphorylase (EC [{{EClink}}2.4.1.8 2.4.1.8]) from ''Lactobacillus sanfranciscensis'' DSM 20451T <cite>Ehrmann1998</cite>
: trehalose phosphorylase (EC [{{EClink}}2.4.1.64 2.4.1.64]) from ''Thermoanaerobacter brockii'' ATCC 35047 <cite> Maruta2002</cite>
: kojibiose phosphorylase (EC [{{EClink}}2.4.1.230 2.4.1.230]) from ''Thermoanaerobacter brockii'' ATCC 35047 <cite>Yamatomo2004</cite>
: trehalose 6-phosphate phosphorylase (EC [{{EClink}}2.4.1.- 2.4.1.-]) from ''Lactococcus lactis'' ssp. ''lactis'' 19435 <cite> Andersson2001</cite>
: α,α-trehalase (EC [{{EClink}}3.2.1.28 3.2.1.28]) from ''Saccharomyces cerevisiae'' S288C <cite>Destruelle1995</cite>.
;First general acid residue identification: maltose phosphorylase (EC [{{EClink}}2.4.1.8 2.4.1.8]) from ''Lactobacillus brevis'' by X-ray structure analysis <cite>Egloff2001</cite> and confirmed by mutagenesis for ''Paenibacillus'' sp. maltose phosphorylase (EC [{{EClink}}2.4.1.8 2.4.1.8]) <cite>Hidaka2005</cite>.
;First three-dimentional structure determination: maltose phosphorylase (EC [{{EClink}}2.4.1.8 2.4.1.8]) from ''Lactobacillus brevis'' <cite>Egloff2001</cite>.

== References ==
<biblio>
#Fitting1952 pmid=12999827
#Nakai2009 pmid=19919544
#Hidaka2004 pmid=15274915
#Tsumuraya1990 pmid=2143366
#Egloff2001 pmid=11587643
#Aleshin1992 pmid=1527049
#Hidaka2005 Hidaka Y, Hatada Y, Akita M, Yoshida M, Nakamura N, Takada M, Nakakuki T, Ito S, and Horikoshi K. ''Maltose phosphorylase from a deep-sea Paenibacillus sp.: Enzymatic properties and nucleotide and amino-acid sequences.'' Enzyme and Microbial Technology, Volume 37, Issue 2, 1 July 2005, Pages 185-194. doi:10.1016/j.enzmictec.2005.02.010
#Hidaka2006 pmid=16646954
#Cantarel2009 pmid=18838391
#Ehrmann1998 pmid=9851037
#Maruta2002 pmid=12400703
#Yamatomo2004 pmid=16233673
#Andersson2001 pmid=11553642
#Destruelle1995 pmid=7502577
</biblio>

[[Category:Glycoside Hydrolase Families|GH065]]

User:Hiroyuki Nakai

2011-05-26T02:19:07Z

Hiroyuki Nakai:

[[Image:Nakai.jpg|thumb|200px|right]]
'''Hiroyuki Nakai''' is an assistant professor at Food Glycoscience Laboratory in Graduate School of Science and Technology, Niigata University. He obtained Ph. D from Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University in 2005, under the supervision of Prof. Atsuo Kimura and Prof. Seiya Chiba. He joined the group of Prof. Birte Svensson at Enzyme and Protein Chemistry, Department of Systems Biology, the Technical University of Denmark as post-doctoral fellow (2007) and assistant professor (2008-2010), working on chemoenzymatic synthesis of oligosaccharides by glycoside hydrolase of [[GH36]] <cite>Nakai2010a</cite> and phosphorylases of [[GH94]] <cite>Nakai2010b</cite> and [[GH65]] <cite>Nakai2010c Nakai2010d Nakai2009</cite> catalysing transglycosylation and reverse phosphorolysis, respectively. His recent research interests concern structure and function of Carbohydrate Active enZymes, enzymatic synthesis of oligosaccharides showing healthful effect on human intestine, antibacterial activity, and immunostimulatory activity.

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<biblio>
#Nakai2010a pmid=20681989
#Nakai2010b pmid=20678539
#Nakai2010c pmid=20713411
#Nakai2010d pmid=20392438
#Nakai2009 pmid=19919544
</biblio>

File:Nakai.jpg

2011-05-26T02:07:58Z

Hiroyuki Nakai:

User:Hiroyuki Nakai

2011-05-25T07:25:00Z

Hiroyuki Nakai: Created page with "'''Hiroyuki Nakai''' is an assistant professor at Food Glycoscience Laboratory in Graduate School of Science and Technology, Niigata University. He obtained Ph. D from Division o..."

'''Hiroyuki Nakai''' is an assistant professor at Food Glycoscience Laboratory in Graduate School of Science and Technology, Niigata University. He obtained Ph. D from Division of Applied Bioscience, Graduate School of Agriculture, Hokkaido University in 2005, under the supervision of Prof. Atsuo Kimura and Prof. Seiya Chiba. He joined the group of Prof. Birte Svensson at Enzyme and Protein Chemistry, Department of Systems Biology, the Technical University of Denmark as post-doctoral fellow (2007) and assistant professor (2008-2010), working on chemoenzymatic synthesis of oligosaccharides by glycoside hydrolase and phosphorylase catalysing transglycosylation and reverse phosphorolysis, respectively. His recent research interests concern structure and function of Carbohydrate Active enZymes, enzymatic synthesis of oligosaccharides showing healthful effect on human intestine, antibacterial activity, and immunostimulatory activity.

* Nakai H., Abou Hachem M., Petersen B.O., Westphal Y., Mannerstedt K., Baumann M.J., Dilokpimol A., Schols H.A., Duus J.Ø., Svensson B. Efficient chemoenzymatic oligosaccharide synthesis by reverse phosphorolysis using cellobiose phosphorylase and cellodextrin phosphorylase from ''Clostridium thermocellum.''　Biochimie (2010) 92, 1818–1826.

* Nakai H., Baumann M.J., Petersen B.O., Westphal Y., Abou Hachem M., Dilokpimol A., Duus J.Ø., Schols H.A., Svensson B.　''Aspergillus nidulans'' α-galactosidase of glycoside hydrolase family 36 catalyses the formation of α-galacto-oligosaccharides by transglycosylation. The FEBS Journal (2010) 277, 3538–3551.

* Nakai H., Petersen B.O., Westphal Y., Dilokpimol A., Abou Hachem M., Duus J.Ø., Schols H.A., Svensson B. Rational engineering of ''Lactobacillus acidophilus'' NCFM maltose phosphorylase into either trehalose or kojibiose dual specificity phosphorylase. Protein Engineering Design & Selection (2010) 23, 781–787.

* Nakai H., Dilokpimol A., Abou Hachem M., Svensson B. Efficient one-pot enzymatic synthesis of α-(1→4)-glucosidic disaccharides through a coupled reaction catalysed by ''Lactobacillus acidophilus'' NCFM maltose phosphorylase. Carbohydrate Research (2010) 345, 1061–1064.

* Nakai H., Baumann M.J., Petersen B.O., Westphal Y., Schols H.A., Dilokpimol A., Abou Hachem M., Lahtinen S.J., Duus J.Ø., Svensson B. The maltodextrin transport system and metabolism in ''Lactobacillus acidophilus'' NCFM and production of novel α-glucosides through reverse phosphorolysis by maltose phosphorylase. The FEBS Journal (2009) 276, 7353–7365.