Additional carbohydrate-binding modules enhance the insoluble substrate-hydrolytic activity of beta-1,3-glucanase from alkaliphilic Nocardiopsis sp. F96.
Beta-1,3-glucanase (BglF) from Nocardiopsis sp. F96 is composed of only a catalytic domain. To improve the enzymatic properties of BglF, we attempted to construct chimeric enzymes consisting of BglF and some carbohydrate-binding modules, such as the C-terminal additional domain (CAD) and the N-terminal additional domain (NAD) of beta-1,3-glucanase H from Bacillus circulans IAM1165 and the chitin-binding domain (ChBD) of chitinase from alkaliphilic Bacillus sp. J813. CAD-fused BglF (BglF-CAD), NAD-fused BglF (NAD-BglF), both NAD- and CAD-fused BglF (NAD-BglF-CAD) and ChBD-fused BglF (BglF-ChBD) were constructed and characterized. The addition of CAD caused increases in binding abilities and hydrolytic activities toward insoluble beta-1,3-glucans. As well as BglF-CAD, the binding ability and hydrolytic activity of BglF-ChBD toward pachyman were also increased. The hydrolytic activity of BglF-CAD at pH 9-10 was higher than that of BglF. The relative activities of BglF-CAD and BglF-ChBD at around 50-70 degrees C were higher than that of BglF.
Koizumi
Naoya
N
Department of Bioengineering, Tokyo Institute of Technology, Yokohama, Japan.
Masuda
Sumiko
S
Maeda
Kiyoe
K
Isoda
Yuya
Y
Yatsunami
Rie
R
Fukui
Toshiaki
T
Nakamura
Satoshi
S
D000192:Actinomycetales / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D001407:Bacillus / Q000201:enzymology; D050260:Carbohydrate Metabolism; D020134:Catalytic Domain; D002417:Cattle; D002686:Chitin / Q000378:metabolism; D002688:Chitinases / Q000737:chemistry / Q000378:metabolism; D043326:Glucan 1,3-beta-Glucosidase / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D013696:Temperature
D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; D002686:Chitin; D002688:Chitinases; D043326:Glucan 1,3-beta-Glucosidase