Dataset Information

A Novel Glycoside Hydrolase Family 5 ?-1,3-1,6-Endoglucanase from Saccharophagus degradans 2-40T and Its Transglycosylase Activity.

ABSTRACT: In this study, we characterized Gly5M, originating from a marine bacterium, as a novel ?-1,3-1,6-endoglucanase in glycoside hydrolase family 5 (GH5) in the Carbohydrate-Active enZyme database. The gly5M gene encodes Gly5M, a newly characterized enzyme from GH5 subfamily 47 (GH5_47) in Saccharophagus degradans 2-40(T) The gly5M gene was cloned and overexpressed in Escherichia coli Through analysis of the enzymatic reaction products by thin-layer chromatography, high-performance liquid chromatography, and matrix-assisted laser desorption ionization-tandem time of flight mass spectrometry, Gly5M was identified as a novel ?-1,3-endoglucanase (EC 3.2.1.39) and bacterial ?-1,6-glucanase (EC 3.2.1.75) in GH5. The ?-1,3-endoglucanase and ?-1,6-endoglucanase activities were detected by using laminarin (a ?-1,3-glucan with ?-1,6-glycosidic linkages derived from brown macroalgae) and pustulan (a ?-1,6-glucan derived from fungal cell walls) as the substrates, respectively. This enzyme also showed transglycosylase activity toward ?-1,3-oligosaccharides when laminarioligosaccharides were used as the substrates. Since laminarin is the major form of glucan storage in brown macroalgae, Gly5M could be used to produce glucose and laminarioligosaccharides, using brown macroalgae, for industrial purposes.In this study, we have discovered a novel ?-1,3-1,6-endoglucanase with a unique transglycosylase activity, namely, Gly5M, from a marine bacterium, Saccharophagus degradans 2-40(T) Gly5M was identified as the newly found ?-1,3-endoglucanase and bacterial ?-1,6-glucanase in GH5. Gly5M is capable of cleaving glycosidic linkages of both ?-1,3-glucans and ?-1,6-glucans. Gly5M also possesses a transglycosylase activity toward ?-1,3-oligosacchrides. Due to the broad specificity of Gly5M, this enzyme can be used to produce glucose or high-value ?-1,3- and/or ?-1,6-oligosaccharides.

SUBMITTER: Wang D

PROVIDER: S-EPMC4959192 | biostudies-literature | 2016 Jul

REPOSITORIES: biostudies-literature

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A Novel Glycoside Hydrolase Family 5 β-1,3-1,6-Endoglucanase from Saccharophagus degradans 2-40T and Its Transglycosylase Activity.

Wang Damao D Kim Do Hyoung DH Seo Nari N Yun Eun Ju EJ An Hyun Joo HJ Kim Jae-Han JH Kim Kyoung Heon KH

Applied and environmental microbiology 20160630 14

<h4>Unlabelled</h4>In this study, we characterized Gly5M, originating from a marine bacterium, as a novel β-1,3-1,6-endoglucanase in glycoside hydrolase family 5 (GH5) in the Carbohydrate-Active enZyme database. The gly5M gene encodes Gly5M, a newly characterized enzyme from GH5 subfamily 47 (GH5_47) in Saccharophagus degradans 2-40(T) The gly5M gene was cloned and overexpressed in Escherichia coli Through analysis of the enzymatic reaction products by thin-layer chromatography, high-performance ...[more]

PMID: 27208098

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Project description:Background:Laminarin is a potential biomass feedstock for the production of glucose, which is the most preferable fermentable sugar in many microorganisms by which it can be converted to biofuels and bio-based chemicals. Also, laminarin is a good resource as functional materials because it consists of ?-1,3-glucosidic linkages in its backbone and ?-1,6-glucosidic linkages in its branches so that its oligosaccharides driven from laminarin have a variety of biological activities. It is industrially important to be able to produce laminarioligosaccharides as well as glucose from laminarin by a single enzyme because the enzyme cost accounts for a large part of bio-based products. In this study, we investigated the industrial applicability of Bgl1B, a unique ?-glucosidase from Saccharophagus degradans 2-40T, belonging to the glycoside hydrolase family 1 (GH1) by characterizing its activity of hydrolyzing laminarin under various conditions. Results:Bgl1B was cloned and overexpressed in Escherichia coli from S. degradans 2-40T, and its enzymatic activity was characterized. Similar to most of ?-glucosidases in GH1, Bgl1B was able to hydrolyze a variety of disaccharides having different ?-linkages, such as laminaribiose, cellobiose, gentiobiose, lactose, and agarobiose, by cleaving ?-1,3-, ?-1,4-, and ?-1,6-glycosidic linkages. However, Bgl1B showed the highest specific activity toward laminaribiose with a ?-1,3-glycosidic linkage. In addition, it was able to hydrolyze laminarin, one of the major polysaccharides in brown macroalgae, into glucose with a conversion yield of 75% of theoretical maximum. Bgl1B also showed transglycosylation activity by producing oligosaccharides from laminarin and laminaribiose under a high mass ratio of substrate to enzyme. Furthermore, Bgl1B was found to be psychrophilic, exhibiting relative activity of 59-85% in the low-temperature range of 2-20 °C. Conclusions:Bgl1B can directly hydrolyze laminarin into glucose with a high conversion yield without leaving any oligosaccharides. Bgl1B can exhibit high enzymatic activity in a broad range of low temperatures (2-20 °C), which is advantageous for establishing energy-efficient bioprocesses. In addition, under high substrate to enzyme ratios, Bgl1B can produce high-value laminarioligosaccharides via its transglycosylation activity. These results show that Bgl1B can be an industrially important enzyme for the production of biofuels and bio-based chemicals from brown macroalgae.

Dataset Information

A Novel Glycoside Hydrolase Family 5 ?-1,3-1,6-Endoglucanase from Saccharophagus degradans 2-40T and Its Transglycosylase Activity.

Publications

A Novel Glycoside Hydrolase Family 5 β-1,3-1,6-Endoglucanase from Saccharophagus degradans 2-40T and Its Transglycosylase Activity.

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