Transglycosylating glycoside hydrolase family 1 β-glucosidase from Penicillium funiculosum NCL1: Heterologous expression in Escherichia coli and characterization

Ramani, Gurusamy; Meera, Balasubramanian; Rajendhran, Jeyaprakash; Gunasekaran, Paramasamy

doi:10.1016/j.bej.2015.03.018

Cited by 8 publications

(4 citation statements)

References 32 publications

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“…2). A comparison of the surface structures (Fig 3) showed that the active sites of Pf Bgl3A and Pf Bgl3B are open and shallow, whereas that of Pf Bgl1A is deep, narrow and tunnel-like consistent with reports on active site configuration for GH1 β-glucosidases (26, 28–31). This active site conformation is thought to improve the glucose tolerance of GH1 proteins by preventing the direct access of glucose to the active site (32).…”

Section: Resultssupporting

confidence: 85%

Profiling of the β-glucosidases identified in the genome ofPenicillium funiculosum: Insights from genomics, transcriptomics, proteomics and homology modelling studies

Okereke¹,

Gupta²,

Ogunyewo³

et al. 2022

Preprint

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Enzymatic lignocellulosic biomass conversion to bioethanol is dependent on efficient enzyme systems with β-glucosidase as a key component. In this study, we performed in-depth profiling of the various β-glucosidases present in the genome of the hypercellulolytic fungus;Penicillium funiculosumusing genomics, transcriptomics, proteomics and molecular dynamics simulation approaches. Of the eight β-glucosidase genes identified in theP. funiculosumgenome, three were found to be extracellular, as evidenced by presence of signal peptides and mass spectrometry. Among the three secreted β-glucosidase, two belonged to the GH3 and one belonged to GH1 families. Modelled structures of these proteins predicted a deep and narrow active site for the GH3 β-glucosidases (PfBgl3A andPfBgl3B) and a shallow open active site for the GH1 β-glucosidase (PfBgl1A). The enzymatic assays indicated thatP. funiculosumsecretome showed high β-glucosidase activities with prominent bands on 4-methylumbelliferyl β-D-glucopyranoside (MUG) zymogram. To understand the contributory effect of each of the three secreted β-glucosidases (PfBgls), the corresponding gene was deleted separately and the effect of the deletion on β-glucosidase activity of the secretome was examined. Although not the most abundant β-glucosidase,PfBgl3A was found to be the most significant one as evidenced by a 42 % reduction in β-glucosidase activity in the ΔPfBgl3A strain. To improve the thermostability, two mutants ofPfBgl3A were designed with the help of molecular dynamics (MD) simulation and were expressed in Pichia pastoris for evaluation. ThePfBgl3A mutant (Mutant A) gave 1.4 fold increase in the half-life (T1/2) of the enzyme at 50 °C.

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Section: Resultssupporting

confidence: 85%

Profiling of the β-glucosidases identified in the genome ofPenicillium funiculosum: Insights from genomics, transcriptomics, proteomics and homology modelling studies

Okereke¹,

Gupta²,

Ogunyewo³

et al. 2022

Preprint

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“…Some β-glucosidases have been shown to have transglycosylation activity which facilitates the synthesis of gentiooligosaccharides. These enzymatic processes have the advantages of strong specificity, mild reaction conditions, high catalytic efficiency, and easy isolation from the product mixture [ 31 , 32 ]. The reported β-glucosidases, using a high concentration of glucose as the substrate, can transfer free glucose to other sugar substrates by forming β-1,6-glycosidic bonds, thereby synthesizing oligosaccharides [ 8 , 33 , 34 ].…”

Section: Resultsmentioning

confidence: 99%

A Novel Neutral and Mesophilic β-Glucosidase from Coral Microorganisms for Efficient Preparation of Gentiooligosaccharides

Zhang

et al. 2021

Foods

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β-glucosidases can produce gentiooligosaccharides that are lucrative and promising for the prebiotic and alternative food industries. However, the commercial production of gentiooligosaccharides using β-glucosidase is challenging, as this process is limited by the need for high thermal energy and increasing demand for the enzyme. Here, a putative β-glucosidase gene, selected from the coral microbial metagenome, was expressed in Escherichia coli. Reverse hydrolysis of glucose by Blg163 at pH 7.0 and 40 °C achieved a gentiooligosaccharide yield of 43.02 ± 3.20 g·L−1 at a conversion rate of 5.38 ± 0.40%. Transglycosylation of mixed substrates, glucose and cellobiose, by Blg163 consumed 21.6 U/0.5 g glucose/g cellobiose, achieving a gentiooligosaccharide yield of 70.34 ± 2.20 g·L−1 at a conversion rate of 15.63%, which is close to the highest yield reported in previous findings. Blg163-mediated synthesis of gentiooligosaccharides is the mildest reaction and the lowest β-glucosidase consumption reported to date.

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“…One such example is a β-glucosidase from Penicillium funiculosum . The recombinantly expressed rBgl6 demonstrated the transglycosylation of glucose to form the β-1,6 linked glucose dimer gentiobiose with a yield of 23% [ 63 ]. The bacterial endo-1,3-β- d -glucanase from Formosa algae is able to perform transglycosylation of the acceptors methyl-β- d -xylopyranoside, methyl-α- d -glucopyranoside, 4-methylumbelliferyl-β-d-glucoside and glycerol, and utilizes laminarin as the donor.…”

Section: Glycoside Hydrolasesmentioning

confidence: 99%

Recent advances in enzymatic synthesis of β-glucan and cellulose

Andrade

Field

et al. 2021

Carbohydrate Research

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Bottom-up synthesis of β-glucans such as callose, fungal β-(1,3)(1,6)-glucan and cellulose, can create the defined compounds that are needed to perform fundamental studies on glucan properties and develop applications. With the importance of β-glucans and cellulose in high-profile fields such as nutrition, renewables-based biotechnology and materials science, the enzymatic synthesis of such relevant carbohydrates and their derivatives has attracted much attention. Here we review recent developments in enzymatic synthesis of β-glucans and cellulose, with a focus on progress made over the last five years. We cover the different types of biocatalysts employed, their incorporation in cascades, the exploitation of enzyme promiscuity and their engineering, and reaction conditions affecting the production as well as in situ self-assembly of (non)functionalised glucans. The recent achievements in the application of glycosyl transferases and β-1,4- and β-1,3-glucan phosphorylases demonstrate the high potential and versatility of these biocatalysts in glucan synthesis in both industrial and academic contexts.

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Transglycosylating glycoside hydrolase family 1 β-glucosidase from Penicillium funiculosum NCL1: Heterologous expression in Escherichia coli and characterization

Cited by 8 publications

References 32 publications

Profiling of the β-glucosidases identified in the genome ofPenicillium funiculosum: Insights from genomics, transcriptomics, proteomics and homology modelling studies

Profiling of the β-glucosidases identified in the genome ofPenicillium funiculosum: Insights from genomics, transcriptomics, proteomics and homology modelling studies

A Novel Neutral and Mesophilic β-Glucosidase from Coral Microorganisms for Efficient Preparation of Gentiooligosaccharides

Recent advances in enzymatic synthesis of β-glucan and cellulose

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