Cloning and expression in <i>Pichia pastoris</i> of a blue mussel  (<i>Mytilus edulis</i>) β‐mannanase gene

Xu, Bingze; Sellos, Daniel; Janson, Jan‐Christer

doi:10.1046/j.1432-1327.2002.02824.x

Cited by 39 publications

(36 citation statements)

References 35 publications

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“…In each case, the GH-A (␤/␣) 8 barrels were predominant in the 3D-jury consensus results with the scores ranging from 74 to 198. The scores were well above the benchmark of 50, suggesting that all GH113 members folded into a GH-A (␤/␣) 8 barrel. Sequence alignment between AaManA and the other GH113 members showed that the catalytic residues (indicated by the diamond and triangle in Fig.…”

Section: Overall Structure Comparison With Other Glycoside Hydrolases-mentioning

confidence: 88%

“…As shown in Fig. 5A, AaManA folds into a typical (␤/␣) 8 -barrel. On the top of the barrel, AaManA features a groove (indicated by a red arrow in Fig.…”

Section: Characterization Of the Gene Cluster Containing Aamana-mentioning

confidence: 92%

“…AaManA adopts the GH-A (␤/␣) 8 barrel and belongs to clan GH-A. Additionally, the possible fold of each of the members assigned to the novel GH113 family was analyzed by the BioInfoBank metaserver.…”

Section: Overall Structure Comparison With Other Glycoside Hydrolases-mentioning

confidence: 99%

“…1, A and B) (4); these enzymes have been isolated from bacteria, fungi, plants, and some mollusks (5)(6)(7)(8). Interest in these enzymes has been increasing due to their importance in hemicellulose hydrolysis and various other applications (5,9,10).…”

mentioning

confidence: 99%

“…At present, the tertiary structures of seven mannanases have been determined, of which five are from GH family 5 (12)(13)(14)(15)(16) and two are from family 26 (17,18). All of these mannanases share a common (␤/␣) 8 barrel fold, a retaining reaction mechanism (Fig. 1C), and two conserved catalytic residues (two glutamate residues at the C termini of ␤4 and ␤7, respectively).…”

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confidence: 99%

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Biochemical and Structural Characterization of the Intracellular Mannanase AaManA of Alicyclobacillus acidocaldarius Reveals a Novel Glycoside Hydrolase Family Belonging to Clan GH-A

Zhang

Peng

et al. 2008

Journal of Biological Chemistry

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An intracellular mannanase was identified from the thermoacidophile Alicyclobacillus acidocaldarius Tc-12-31. This enzyme is particularly interesting, because it shows no significant sequence similarity to any known glycoside hydrolase. Gene cloning, biochemical characterization, and structural studies of this novel mannanase are reported in this paper. The gene consists of 963 bp and encodes a 320-amino acid protein, AaManA. Based on its substrate specificity and product profile, AaManA is classified as an endo-␤-1,4-mannanase that is capable of transglycosylation. Kinetic analysis studies revealed that the enzyme required at least five subsites for efficient hydrolysis. The crystal structure at 1.9 Å resolution showed that AaManA adopted a (␤/␣) 8 -barrel fold. Two catalytic residues were identified: Glu 151 at the C terminus of ␤-stand ␤4 and Glu 231 at the C terminus of ␤7. Based on the structure of the enzyme and evidence of its transglycosylation activity, AaManA is placed in clan GH-A. Superpositioning of its structure with that of other clan GH-A enzymes revealed that six of the eight GH-A key residues were functionally conserved in AaManA, with the exceptions being residues Thr 95 and Cys 150 . We propose a model of substrate binding in AaManA in which Glu 282 interacts with the axial OH-C(2) in ؊2 subsites. Based on sequence comparisons, the enzyme was assigned to a new glycoside hydrolase family (GH113) that belongs to clan GH-A.Mannans are polysaccharides found in plants and consist of a backbone of ␤-1,4-linked mannose and glucose units. Mannose residues often carry an ␣-galactosyl substitute at O-6, and the degree of substitution depends on the plant of origin (Fig. 1A) (1). Mannans are widely distributed in nature in parts of the hemicellulose fraction in hardwoods and softwoods (1), legume seeds (2), and beans of carob trees (3).Endo-␤-1,4-mannanases (mannanases; EC 3.2.1.78) are glycoside hydrolases that randomly cleave the ␤-1,4-linkage in mannans (Fig. 1, A and B) (4); these enzymes have been isolated from bacteria, fungi, plants, and some mollusks (5-8). Interest in these enzymes has been increasing due to their importance in hemicellulose hydrolysis and various other applications (5, 9, 10). During the last 2 decades, more than 80 sequences of the catalytic domains of mannanase have been reported (see the CAZy site on the World Wide Web) and classified into glycoside hydrolase (GH) 2 families 5 and 26, based on their sequence similarities (11). In recent years, crystallographic resolution of the structures of these enzymes has yielded information on their structure. At present, the tertiary structures of seven mannanases have been determined, of which five are from GH family 5 (12-16) and two are from family 26 (17, 18). All of these mannanases share a common (␤/␣) 8 barrel fold, a retaining reaction mechanism (Fig. 1C), and two conserved catalytic residues (two glutamate residues at the C termini of ␤4 and ␤7, respectively). Therefore, all of these enzymes have been assigned to the same GH ...

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Section: Overall Structure Comparison With Other Glycoside Hydrolases-mentioning

confidence: 88%

“…As shown in Fig. 5A, AaManA folds into a typical (␤/␣) 8 -barrel. On the top of the barrel, AaManA features a groove (indicated by a red arrow in Fig.…”

Section: Characterization Of the Gene Cluster Containing Aamana-mentioning

confidence: 92%

Section: Overall Structure Comparison With Other Glycoside Hydrolases-mentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Biochemical and Structural Characterization of the Intracellular Mannanase AaManA of Alicyclobacillus acidocaldarius Reveals a Novel Glycoside Hydrolase Family Belonging to Clan GH-A

Zhang

Peng

et al. 2008

Journal of Biological Chemistry

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Cloning, expression and characterization of endo‐β‐1,4‐mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris

Duruksu

Öztürk

Biely

et al. 2009

Biotechnology Progress

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in Wiley InterScience (www.interscience.wiley.com).To be utilized in biomass conversion, including ethanol production and galactosylated oligosaccharide synthesis, namely prebiotics, the gene of extracellular endo-b-1,4-mannanase (EC 3.2.1.78) of Aspergillus fumigatus IMI 385708 (formerly known as Thermomyces lanuginosus IMI 158749) was expressed first in Aspergillus sojae and then in Pichia pastoris under the control of the glyceraldehyde triphosphate dehydrogenase (gpdA) and the alcohol oxidase (AOX1) promoters, respectively. The highest production of mannanase (352 U mL À1 ) in A. sojae was observed after 6 days of cultivation. In P. pastoris, the highest mannanase production was observed 10 h after induction with methanol (61 U mL À1 ). The fold increase in mannanase production was estimated as $12-fold and $2-fold in A. sojae and P. pastoris, respectively, when compared with A. fumigatus. Both recombinant enzymes showed molecular mass of about 60 kDa and similar specific activities ($350 U mg À1 protein). Temperature optima were at 60 C and 45 C, and maximum activity was at pH 4.5 and 5.2 for A. sojae and P. pastoris, respectively. The enzyme from P. pastoris was more stable retaining most of the activity up to 50 C, whereas the enzyme from A. sojae rapidly lost activity above 40 C.

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Mannanase

Huerta‐Ochoa¹,

Prado-Barragán²,

Gracida³

et al. 2006

Enzyme Technology

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Cloning and expression in Pichia pastoris of a blue mussel (Mytilus edulis) β‐mannanase gene

Cited by 39 publications

References 35 publications

Biochemical and Structural Characterization of the Intracellular Mannanase AaManA of Alicyclobacillus acidocaldarius Reveals a Novel Glycoside Hydrolase Family Belonging to Clan GH-A

Biochemical and Structural Characterization of the Intracellular Mannanase AaManA of Alicyclobacillus acidocaldarius Reveals a Novel Glycoside Hydrolase Family Belonging to Clan GH-A

Cloning, expression and characterization of endo‐β‐1,4‐mannanase from Aspergillus fumigatus in Aspergillus sojae and Pichia pastoris

Mannanase

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