Novel glucose dehydrogenase from Mucor prainii: Purification, characterization, molecular cloning and gene expression in Aspergillus sojae

Satake, Ryoko; Ichiyanagi, Atsushi; Ichikawa, Keiichi; Hirokawa, Kozo; Araki, Yasuko; Yoshimura, Taro; Gomi, Keiko

doi:10.1016/j.jbiosc.2015.03.012

Cited by 13 publications

(6 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Even with structural and chemical information about the active center pocket obtained by crystal structure analysis of the enzyme, it would be difficult to predict the suitable mediator to use. Although there is an increasing number of reports on the discovery and recombinant production of new FAD-GDHs from various fungi [20,21,22,23,24,25,26], it is not realistic from the viewpoint of time and cost to clarify the mediator structure for all these newly discovered enzymes. Therefore, in this study, we propose a strategy for determining the optimum mediator for FAD-GDHs by screening a set of organic redox mediators via their bimolecular rate constants for the enzyme reaction.…”

Section: Introductionmentioning

confidence: 99%

Bimolecular Rate Constants for FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus and Organic Electron Acceptors

Tsuruoka

Sadakane

Hayashi

et al. 2017

IJMS

View full text Add to dashboard Cite

Abstract:The flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH) from Aspergillus species require suitable redox mediators to transfer electrons from the enzyme to the electrode surface for the application of bioelectrical devices. Although several mediators for FAD-GDH are already in use, they are still far from optimum in view of potential, kinetics, sustainability, and cost-effectiveness. Herein, we investigated the efficiency of various phenothiazines and quinones in the electrochemical oxidation of FAD-GDH from Aspergillus terreus. At pH 7.0, the logarithm of the bimolecular oxidation rate constants appeared to depend on the redox potentials of all the mediators tested. Notably, the rate constant of each molecule for FAD-GDH was approximately 2.5 orders of magnitude higher than that for glucose oxidase from Aspergillus sp. The results suggest that the electron transfer kinetics is mainly determined by the formal potential of the mediator, the driving force of electron transfer, and the electron transfer distance between the redox active site of the mediator and the FAD, affected by the steric or chemical interactions. Higher k 2 values were found for ortho-quinones than for para-quinones in the reactions with FAD-GDH and glucose oxidase, which was likely due to less steric hindrance in the active site in the case of the ortho-quinones.

show abstract

Section: Introductionmentioning

confidence: 99%

Bimolecular Rate Constants for FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus and Organic Electron Acceptors

Tsuruoka

Sadakane

Hayashi

et al. 2017

IJMS

View full text Add to dashboard Cite

show abstract

“…In order to test the above hypotheses, we employed the A. sojae pyrG marker (Satake et al, 2015) encoding orothidine-5'-phosphate decarboxylase, since its orthologs are used for increasing the plasmid copy number in yeast (Loison et al,1989;Verma et al, 2012) and the background growth of pyrG-defective strains in various filamentous fungi is very low on a medium without uridine/uracil. In A. sojae, the one-copy integration of pyrG with a promoter of 407 bp (pyrG407) could effectively complement the growth defect of a pyrG-defective strain on…”

Section: Discussionmentioning

confidence: 99%

“…An equal amount (50 mg) of mycelium of each strain was frozen in liquid nitrogen and treated with beads cell disrupter MS-100R (Tomy Seiko Co., Ltd., Japan), and then suspended in 1 mL of a 100 mM potassium phosphate buffer (pH 7.0). GDH activity was assayed in the soluble fraction of homogenates according to a method described previously (Satake et al, 2015). GDH activity was undetectable in the host strain and not influenced by Asgdh disruption, and, thus, the relative GDH activity of each transformant was calculated by comparison with the GDH activity of As-p407-1CP.…”

Section: Discussionmentioning

confidence: 99%

“…3 Table Number and The glucose dehydrogenase gene (Mpgdh) is derived from Mucor prainii, and the translation elongation factor promoter (Ptef, 748 bp) and the alkaline protease terminator sequence (Talp, 800 bp) are from A. sojae (Satake et al, 2015). The expression cassette, including Mpgdh and a selection marker variant of pyrG or adeA, was inserted into the multiple cloning site (MCS) of the pUC19 vector (Takara Bio Inc., Shiga, Japan).…”

Section: Tablesmentioning

confidence: 99%

See 1 more Smart Citation

High-level heterologous protein production using an attenuated selection marker in <i>Aspergillus sojae</i>

Araki

Yuzuki

Masakari

et al. 2021

J. Gen. Appl. Microbiol.

Self Cite

View full text Add to dashboard Cite

“…2 In this type of glucose sensor, electron is transferred from FAD as the primary electron acceptor to an external electron acceptor except for molecular oxygen during the enzymatic oxidation of D-glucose. Several studies on FADGDHs from filamentous fungi, including Aspergillus oryzae, [4][5][6] Aspergillus terreus, [7][8][9] Aspergillus flavus, 10,11 Aspergillus niger, 10 Glomerella cingulata, 12,13 and Mucor prainii, 14 have been reported. These FADGDHs have suitable enzymatic properties for glucose sensors.…”

Section: Introductionmentioning

confidence: 99%

Thermostable FAD-dependent Glucose Dehydrogenases from Thermophilic Filamentous Fungus <i>Thermoascus aurantiacus</i>

et al. 2016

View full text Add to dashboard Cite

We newly identified FAD-dependent glucose dehydrogenase (FADGDH) gene homologs from thermophilic filamentous fungus Thermoascus aurantiacus. The gene homologs were cloned from two strains of Th. aurantiacus, NBRC 6766 and NBRC 9748. Recombinant FADGDHs of the two strains were prepared by using Escherichia coli and Pichia pastoris as hosts. Absorption spectra and enzymatic characterization clearly showed that these enzymes contained oxidized FAD as a coenzyme and exhibited glucose dehydrogenase activity. Analysis of thermal stability revealed that the denaturation midpoints (T m ) of these FADGDHs were at least above 71.4°C. The results showed that these FADGDHs exhibited remarkably high thermal stability. We also performed bioelectrochemical experiments using unglycosylated and glycosylated FADGDHs of Th. aurantiacus NBRC 6766, which exhibited higher affinities for glucose than those of Th. aurantiacus NBRC 9748 FADGDHs. Th. aurantiacus NBRC 6766 FADGDHimmobilized electrodes effectively showed current responses to glucose. Therefore, these thermostable FADGDHs could be applied to a bioelectro-catalyst for glucose oxidation with long-term storage and continuous use.

show abstract

Novel glucose dehydrogenase from Mucor prainii: Purification, characterization, molecular cloning and gene expression in Aspergillus sojae

Cited by 13 publications

References 24 publications

Bimolecular Rate Constants for FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus and Organic Electron Acceptors

Bimolecular Rate Constants for FAD-Dependent Glucose Dehydrogenase from Aspergillus terreus and Organic Electron Acceptors

High-level heterologous protein production using an attenuated selection marker in <i>Aspergillus sojae</i>

Thermostable FAD-dependent Glucose Dehydrogenases from Thermophilic Filamentous Fungus <i>Thermoascus aurantiacus</i>

Contact Info

Product

Resources

About