Dataset Information

Expression and Characteristics of Two Glucose-Tolerant GH1 ?-glucosidases From Actinomadura amylolytica YIM 77502T for Promoting Cellulose Degradation.

ABSTRACT: The bioconversion of lignocellulose in various industrial processes, such as biofuel production, requires the degradation of cellulose. Actinomadura amylolytica YIM 77502T is an aerobic, Gram-positive actinomycete that can efficiently degrade crystalline cellulose by extracellular cellulases. Genomic analysis of A. amylolytica identified 9 cellulase and 11 ?-glucosidase genes that could potentially encode proteins that digest cellulose. Extracellular proteome characterization of A. amylolytica cell-free culture supernatant by liquid chromatography tandem mass spectrometry analysis revealed that 4 of these cellulases and 2 of these ?-glucosidases functioned during cellulose hydrolysis. Thin-layer chromatography analysis revealed extracellular ?-glucosidases play a major role in carboxyl methyl cellulose (CMC) degradation of products in culture supernatants. In this study, 2 of the identified secreted ?-glucosidases, AaBGL1 and AaBGL2, were functionally expressed in Escherichia coli and found to have ?-glucosidase activity with wide substrate specificities, including for p-nitrophenyl ?-D-glucopyranoside (pNPG), p-nitrophenyl-beta-D-cellobioside (pNPC), and cellobiose. Moreover, AaBGL1 and AaBGL2 had high tolerances for glucose. After adding these ?-glucosidases to commercial cellulases, the degradation rates of CMC, Avicel, birch sawdust, and corncob powder increased by 37, 42, 33, and 9%, respectively. Overall, this work identifies an alternative potential source of ?-glucosidases with potential applications in commercial cellulose utilization and the bioenergy industry.

SUBMITTER: Yin YR

PROVIDER: S-EPMC6305311 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Publications

Expression and Characteristics of Two Glucose-Tolerant GH1 β-glucosidases From Actinomadura amylolytica YIM 77502T for Promoting Cellulose Degradation.

Yin Yi-Rui YR Sang Peng P Xian Wen-Dong WD Li Xin X Jiao Jian-Yu JY Liu Lan L Hozzein Wael N WN Xiao Min M Li Wen-Jun WJ

Frontiers in microbiology 20181218

The bioconversion of lignocellulose in various industrial processes, such as biofuel production, requires the degradation of cellulose. Actinomadura amylolytica YIM 77502T is an aerobic, Gram-positive actinomycete that can efficiently degrade crystalline cellulose by extracellular cellulases. Genomic analysis of A. amylolytica identified 9 cellulase and 11 β-glucosidase genes that could potentially encode proteins that digest cellulose. Extracellular proteome characteriz ...[more]

PMID: 30619214

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Project description:?-glucosidases are key enzymes used in second-generation biofuel production. They act in the last step of the lignocellulose saccharification, converting cellobiose in glucose. However, most of the ?-glucosidases are inhibited by high glucose concentrations, which turns it a limiting step for industrial production. Thus, ?-glucosidases have been targeted by several studies aiming to understand the mechanism of glucose tolerance, pH and thermal resistance for constructing more efficient enzymes. In this paper, we present a database of ?-glucosidase structures, called Glutant?ase. Our database includes 3842 GH1 ?-glucosidase sequences collected from UniProt. We modeled the sequences by comparison and predicted important features in the 3D-structure of each enzyme. Glutant?ase provides information about catalytic and conserved amino acids, residues of the coevolution network, protein secondary structure, and residues located in the channel that guides to the active site. We also analyzed the impact of beneficial mutations reported in the literature, predicted in analogous positions, for similar enzymes. We suggested these mutations based on six previously described mutants that showed high catalytic activity, glucose tolerance, or thermostability (A404V, E96K, H184F, H228T, L441F, and V174C). Then, we used molecular docking to verify the impact of the suggested mutations in the affinity of protein and ligands (substrate and product). Our results suggest that only mutations based on the H228T mutant can reduce the affinity for glucose (product) and increase affinity for cellobiose (substrate), which indicates an increment in the resistance to product inhibition and agrees with computational and experimental results previously reported in the literature. More resistant ?-glucosidases are essential to saccharification in industrial applications. However, thermostable and glucose-tolerant ?-glucosidases are rare, and their glucose tolerance mechanisms appear to be related to multiple and complex factors. We gather here, a set of information, and made predictions aiming to provide a tool for supporting the rational design of more efficient ?-glucosidases. We hope that Glutant?ase can help improve second-generation biofuel production. Glutant?ase is available at http://bioinfo.dcc.ufmg.br/glutantbase .

Dataset Information

Expression and Characteristics of Two Glucose-Tolerant GH1 ?-glucosidases From Actinomadura amylolytica YIM 77502T for Promoting Cellulose Degradation.

Publications

Expression and Characteristics of Two Glucose-Tolerant GH1 β-glucosidases From <i>Actinomadura amylolytica</i> YIM 77502<sup>T</sup> for Promoting Cellulose Degradation.

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