Gene Cloning of Xylanase Glycoside Hydrolase Family 11 from Bacillus halodurans CM1 in Escherichia coli DH5α

2019 ◽  
Vol 13 (4) ◽  
pp. 123-128
Author(s):  
MUHAMAD TAUFIQUL NAUFAL ◽  
◽  
AGUSTIN KRISNA WARDANI ◽  
IS HELIANTI ◽  
◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Gene Cloning ◽  
Glycoside Hydrolase ◽  
Bacillus Halodurans ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family

Cloning, Expression and Characterization of a Glycoside Hydrolase Family 39 Xylosidase from Bacillus Halodurans C-125

2007 ◽  
pp. 189-198
Author(s):  
Kurt Wagschal ◽  
Diana Franqui-Espiet ◽  
Charles C. Lee ◽  
George H. Robertson ◽  
Dominic W. S. Wong
Keyword(s):  
Glycoside Hydrolase ◽  
Bacillus Halodurans ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family

scholarly journals Conversion of levoglucosan into glucose by the coordination of four enzymes through oxidation, elimination, hydration, and reduction

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Yuya Kuritani ◽  
Kohei Sato ◽  
Hideo Dohra ◽  
Seiichiro Umemura ◽  
Motomitsu Kitaoka ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Recombinant Proteins ◽  
Metabolic Pathway ◽  
Glycoside Hydrolase ◽  
Gene Locus ◽  
Glycoside Hydrolase Family ◽  
Sequential Reaction ◽  
Layer Chromatography ◽  
Hydrolysis Of ◽  
Hydrolase Family

AbstractLevoglucosan (LG) is an anhydrosugar produced through glucan pyrolysis and is widely found in nature. We previously isolated an LG-utilizing thermophile, Bacillus smithii S-2701M, and suggested that this bacterium may have a metabolic pathway from LG to glucose, initiated by LG dehydrogenase (LGDH). Here, we completely elucidated the metabolic pathway of LG involving three novel enzymes in addition to LGDH. In the S-2701M genome, three genes expected to be involved in the LG metabolism were found in the vicinity of the LGDH gene locus. These four genes including LGDH gene (lgdA, lgdB1, lgdB2, and lgdC) were expressed in Escherichia coli and purified to obtain functional recombinant proteins. Thin layer chromatography analyses of the reactions with the combination of the four enzymes elucidated the following metabolic pathway: LgdA (LGDH) catalyzes 3-dehydrogenation of LG to produce 3-keto-LG, which undergoes β-elimination of 3-keto-LG by LgdB1, followed by hydration to produce 3-keto-d-glucose by LgdB2; next, LgdC reduces 3-keto-d-glucose to glucose. This sequential reaction mechanism resembles that proposed for an enzyme belonging to glycoside hydrolase family 4, and results in the observational hydrolysis of LG into glucose with coordination of the four enzymes.

Cloning, Expression and Characterization of a Glycoside Hydrolase Family 39 Xylosidase from Bacillus Halodurans C-125

2007 ◽  
Vol 146 (1-3) ◽  
pp. 69-78 ◽  
Author(s):  
Kurt Wagschal ◽  
Diana Franqui-Espiet ◽  
Charles C. Lee ◽  
George H. Robertson ◽  
Dominic W. S. Wong
Keyword(s):  
Glycoside Hydrolase ◽  
Bacillus Halodurans ◽  
Glycoside Hydrolase Family ◽  
Hydrolase Family

scholarly journals A New β-Galactosidase from the Antarctic Bacterium Alteromonas sp. ANT48 and Its Potential in Formation of Prebiotic Galacto-Oligosaccharides

Marine Drugs ◽  
2019 ◽  
Vol 17 (11) ◽  
pp. 599 ◽  
Author(s):  
Li ◽  
Zhu ◽  
Xing
Keyword(s):  
Escherichia Coli ◽  
Molecular Docking ◽  
Glycoside Hydrolase ◽  
Intestinal Flora ◽  
Glycoside Hydrolase Family ◽  
Initial Activity ◽  
Antarctic Bacterium ◽  
Encoding Gene ◽  
The Antarctic ◽  
Hydrolase Family

As an important medical enzyme, β-galactosidases catalyze transgalactosylation to form prebiotic Galacto-Oligosaccharides (GOS) that assist in improving the effect of intestinal flora on human health. In this study, a new glycoside hydrolase family 2 (GH2) β-galactosidase-encoding gene, galA, was cloned from the Antarctic bacterium Alteromonas sp. ANT48 and expressed in Escherichia coli. The recombinant β-galactosidase GalA was optimal at pH 7.0 and stable at pH 6.6–7.0, which are conditions suitable for the dairy environment. Meanwhile, GalA showed most activity at 50 °C and retained more than 80% of its initial activity below 40 °C, which makes this enzyme stable in normal conditions. Molecular docking with lactose suggested that GalA could efficiently recognize and catalyze lactose substrates. Furthermore, GalA efficiently catalyzed lactose degradation and transgalactosylation of GOS in milk. A total of 90.6% of the lactose in milk could be hydrolyzed within 15 min at 40 °C, and the GOS yield reached 30.9%. These properties make GalA a good candidate for further applications.

scholarly journals Discovery of solabiose phosphorylase and its application for enzymatic synthesis of solabiose from sucrose and lactose

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Wataru Saburi ◽  
Takanori Nihira ◽  
Hiroyuki Nakai ◽  
Motomitsu Kitaoka ◽  
Haruhide Mori
Keyword(s):  
Escherichia Coli ◽  
Enzymatic Synthesis ◽  
Glycoside Hydrolase ◽  
Ternary Complex ◽  
Glycoside Hydrolase Family ◽  
Efficient Production ◽  
Sucrose Phosphorylase ◽  
Catalytic Activities ◽  
The Past ◽  
Hydrolase Family

AbstractGlycoside phosphorylases (GPs), which catalyze the reversible phosphorolysis of glycosides, are promising enzymes for the efficient production of glycosides. Various GPs with new catalytic activities are discovered from uncharacterized proteins phylogenetically distant from known enzymes in the past decade. In this study, we characterized Paenibacillus borealis PBOR_28850 protein, belonging to glycoside hydrolase family 94. Screening of acceptor substrates for reverse phosphorolysis, in which α-d-glucose 1-phosphate was used as the donor substrate, revealed that the recombinant PBOR_28850 produced in Escherichia coli specifically utilized d-galactose as an acceptor and produced solabiose (β-d-Glcp-(1 → 3)-d-Gal). This indicates that PBOR_28850 is a new GP, solabiose phosphorylase. PBOR_28850 catalyzed the phosphorolysis and synthesis of solabiose through a sequential bi-bi mechanism involving the formation of a ternary complex. The production of solabiose from lactose and sucrose has been established. Lactose was hydrolyzed to d-galactose and d-glucose by β-galactosidase. Phosphorolysis of sucrose and synthesis of solabiose were then coupled by adding sucrose, sucrose phosphorylase, and PBOR_28850 to the reaction mixture. Using 210 mmol lactose and 280 mmol sucrose, 207 mmol of solabiose was produced. Yeast treatment degraded the remaining monosaccharides and sucrose without reducing solabiose. Solabiose with a purity of 93.7% was obtained without any chromatographic procedures.

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