Stabilization of quaternary structure of water-soluble quinoprotein glucose dehydrogenase.

Abstract:

:Water-soluble quinoprotein glucose dehydrogenase (PQQGDH-B) is a dimeric enzyme whose application for glucose sensing is the focus of much attention. We attempted to increase the thermal stability of PQQGDH-B by introducing a disulfide bond at the dimer interface. The Ser residue at position 415 was selected for substitution with Cys, as structural information revealed that its side chains face each other at the dimer interface of PQQGDH-B. PQQGDH-B with Ser415Cys showed 30-fold greater thermal stability at 55 degrees C than did the wild-type enzyme without any decrease in catalytic activity. After incubation at 70 degrees C for 10 min, Ser415Cys retained 90% of the GDH activity of the wild-type enzyme. Disulfide bond formation between the mutant subunits was confirmed by analyses with sodium dodecylsulfate-polyacrylamide gel electrophoresis in the presence and absence of reductants. Our results indicate that the introduction of one Cys residue in each monomer of PQQGDH-B resulted in formation of a disulfide bond at the dimer interface and thus achieved a large increase in the thermal stability of the enzyme.

journal_name

Mol Biotechnol

journal_title

Molecular biotechnology

authors

Igarashi S,Sode K

doi

10.1385/MB:24:2:97

keywords:

subject

Has Abstract

pub_date

2003-06-01 00:00:00

pages

97-104

issue

2

eissn

1073-6085

issn

1559-0305

pii

MB:24:2:97

journal_volume

24

pub_type

杂志文章
  • Cloning, Production and Characterization of a Glycoside Hydrolase Family 7 Enzyme from the Gut Microbiota of the Termite Coptotermes curvignathus.

    abstract::Coptotermes curvignathus is a termite that, owing to its ability to digest living trees, serves as a gold mine for robust industrial enzymes. This unique characteristic reflects the presence of very efficient hydrolytic enzyme systems including cellulases. Transcriptomic analyses of the gut of C. curvignathus revealed...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-017-0015-x

    authors: Woon JS,King PJH,Mackeen MM,Mahadi NM,Wan Seman WMK,Broughton WJ,Abdul Murad AM,Abu Bakar FD

    更新日期:2017-07-01 00:00:00

  • High Conversion of D-Fructose into D-Allulose by Enzymes Coupling with an ATP Regeneration System.

    abstract::D-Allulose is a rare monosaccharide that exists in extremely small quantities in nature, and it is also hard to prepare at a large scale via chemical or enzyme synthetic route due to low conversion and downstream separation complexity. Using D-psicose epimerase and L-rhamnulose kinase, a method enabling high conversio...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-019-00174-6

    authors: Xiao Q,Niu J,Liu H,Liu Y,Zhou X

    更新日期:2019-06-01 00:00:00

  • Methods for construction of adenovirus vectors.

    abstract::Adenoviruses are attracting increasing attention as general purpose mammalian cell expression vectors, as recombinant vaccines, and potentially as vectors for gene therapy. Not only is the adenovirus genome relatively easy to manipulate by recombinant DNA techniques, but adenovirus vectors are relatively stable, grow ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/BF02789331

    authors: Graham FL,Prevec L

    更新日期:1995-06-01 00:00:00

  • Oligovalent Fab display on M13 phage improved by directed evolution.

    abstract::Efficient display of antibody on filamentous phage M13 coat is crucial for successful biopanning selections. We applied a directed evolution strategy to improve the oligovalent display of a poorly behaving Fab fragment fused to phage gene-3 for minor coat protein (g3p). The Fab displaying clones were enriched from a r...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-009-9231-3

    authors: Huovinen T,Sanmark H,Ylä-Pelto J,Vehniäinen M,Lamminmäki U

    更新日期:2010-03-01 00:00:00

  • A highly efficient method for construction of rice artificial MicroRNA vectors.

    abstract::Artificial microRNA (amiRNA) has become a powerful tool for gene silencing in plants. A new method for easy and rapid construction of rice artificial miRNA vector is described. The procedure involved modification of the pCAMBIA1300-UR vector by insertion of a 'vector modification fragment'. This was prepared from the ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-010-9291-4

    authors: Wang X,Yang Y,Yu C,Zhou J,Cheng Y,Yan C,Chen J

    更新日期:2010-11-01 00:00:00

  • Deciphering enzyme function using peptide arrays.

    abstract::Enzymes are key molecules in signal-transduction pathways. However, only a small fraction of more than 500 human kinases, 300 human proteases and 200 human phosphatases is characterised so far. Peptide microarray based technologies for extremely efficient profiling of enzyme substrate specificity emerged in the last y...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/s12033-011-9402-x

    authors: Thiele A,Stangl GI,Schutkowski M

    更新日期:2011-11-01 00:00:00

  • Potential DNA Vaccine for Haemorrhagic Septiceamia Disease.

    abstract::Pasteurella multocida is the main cause of haemorrhagic septicaemia (HS) outbreak in livestock, such as cattle and buffaloes. Conventional vaccines such as alum-precipitated or oil-adjuvant broth bacterins were injected subcutaneously to provide protection against HS. However, the immunity developed is only for short ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-020-00244-0

    authors: Chelliah S,Velappan RD,Lim KT,Swee CWK,Nor Rashid N,Rothan HA,Kabir N,Ismail S

    更新日期:2020-05-01 00:00:00

  • Carotenoids and flavonoids contribute to nutritional protection against skin damage from sunlight.

    abstract::The concept of photoprotection by dietary means is gaining momentum. Plant constituents such as carotenoids and flavonoids are involved in protection against excess light in plants and contribute to the prevention of UV damage in humans. As micronutrients, they are ingested with the diet and are distributed into light...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/s12033-007-0051-z

    authors: Stahl W,Sies H

    更新日期:2007-09-01 00:00:00

  • PVY-resistant transgenic potato plants expressing an anti-NIa protein scFv antibody.

    abstract::A synthetic gene encoding a single chain Fv fragment of an antibody directed against the nuclear inclusion a (NIa) protein of potato virus Y (PVY) was used to transform two commercial potato cultivars (Claustar and BF15). The NIa protease forms the nuclear inclusion body A and acts as the major protease in the cleavag...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:33:2:133

    authors: Gargouri-Bouzid R,Jaoua L,Rouis S,Saïdi MN,Bouaziz D,Ellouz R

    更新日期:2006-06-01 00:00:00

  • Brn-3a, a neuronal transcription factor of the POU gene family: indications for its involvement in cancer and angiogenesis.

    abstract::Brn-3a, a member of the POU gene family (so-called because of the similarity with the group of transcription factors Pit, Oct, and Unc), was found in neuronal cells engaged in the transcription activity of the p1 and p2 promoters of the most powerful antiapoptotic gene, namely, Bcl-2. The alternative splicing of Brn-3...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1385/MB:22:2:123

    authors: Chiarugi V,Del Rosso M,Magnelli L

    更新日期:2002-10-01 00:00:00

  • Gene transfer by electroporation.

    abstract::Electroporation of cells in the presence of DNA is widely used for the introduction of transgenes either stably or transiently into bacterial, fungal, animal, and plant cells. A review of the literature shows that electroporation parameters are often reported in an incomplete or incorrect manner, forcing researchers t...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1007/BF02821542

    authors: Lurquin PF

    更新日期:1997-02-01 00:00:00

  • Surface display of a bifunctional glutathione synthetase on Saccharomyces cerevisiae for converting chicken feather hydrolysate into glutathione.

    abstract::The low economic profits of feather recycling lead that the large amount of feathers is currently discarded in China. To convert feather hydrolysates into GSH with high values, surface display of the bifunctional glutathione synthetase encoded by gcsgs from Streptococcus thermophilus on Saccharomyces cerevisiae and th...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-014-9750-4

    authors: Qiu Z,Tan H,Zhou S,Cao L

    更新日期:2014-08-01 00:00:00

  • Development of a new method for genetic transformation of the green alga Chlorella ellipsoidea.

    abstract::Chlorella ellipsoidea is a single-celled eukaryotic green microalgae with high nutritional value. Its value may be further increased if a simple, reliable and cost-effective transformation method for C. ellipsoidea can be developed. In this paper, we describe a novel transformation method for C. ellipsoidea . This sys...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-012-9554-3

    authors: Liu L,Wang Y,Zhang Y,Chen X,Zhang P,Ma S

    更新日期:2013-06-01 00:00:00

  • Investigation and Expression of the Secoisolariciresinol Dehydrogenase Gene Involved in Podophyllotoxin Biosynthesis.

    abstract::Podophyllotoxin (PPT) is a plant natural product that serves as a precursor for the synthesis of many well-known chemotherapeutic drugs. The limited availability and high demand for the source plants of PPT have led to the exploration of alternative sources for this compound. In this study, we utilized the endophytic ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-015-9888-8

    authors: Arneaud SL,Porter JR

    更新日期:2015-12-01 00:00:00

  • Studying the effects of actin cytoskeletal destabilization on cell cycle by cofilin overexpression.

    abstract::The significance of actin cytoskeleton on cell growth was historically studied using toxic drugs, such as cytochalasin. However, it is possible that unpredictable effects of these agents may have influenced the reported observations. In our study, we have established a drug-free system using cofilin overexpression to ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:31:1:001

    authors: Lee YJ,Keng PC

    更新日期:2005-09-01 00:00:00

  • Transcriptional Study of the RsmZ-sRNAs and Their Relationship to the Biosynthesis of Alginate and Alkylresorcinols in Azotobacter vinelandii.

    abstract::The GacS/A system in Azotobacter vinelandii regulates alginate and alkylresorcinols production through RsmZ1, a small regulatory RNA (sRNA) that releases the translational repression of the algD and arpR mRNAs caused by the RsmA protein. In the Pseudomonadaceae family, the Rsm-sRNAs are grouped into three families: Rs...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-018-0102-7

    authors: López-Pliego L,García-Ramírez L,Cruz-Gómez EA,Domínguez-Ojeda P,López-Pastrana A,Fuentes-Ramírez LE,Núñez C,Castañeda M

    更新日期:2018-09-01 00:00:00

  • Cloning of Toluene 4-Monooxygenase Genes and Application of Two-Phase System to the Production of the Anticancer Agent, Indirubin.

    abstract::Indirubin is a strong inhibitor of several eukaryotic cell signaling pathways and shows promise as a treatment for myelocytic leukemia and Alzheimer's disease. The tmoABCDEF operon, encoding the components of a novel toluene 4-monooxygenase from the paint factory soil isolate, Pseudomonas sp. M4, was cloned and expres...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-015-9863-4

    authors: Wongsaroj L,Sallabhan R,Dubbs JM,Mongkolsuk S,Loprasert S

    更新日期:2015-08-01 00:00:00

  • Isolation of differential genes in suspension cultures of Taxus cuspidata induced by additional taxol.

    abstract::Addition of taxol into suspension cultures of Taxus cuspidata induced cell apoptosis, which was confirmed by gel electrophoresis of the DNA ladders indicating the progressive delineation of fragmented nuclear DNA (nDNA) into distinct bodies. The additional taxol not only changed the microtubule assembly of cells, but ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:20:2:137

    authors: Yuan YJ,Ma ZY,Wu JC

    更新日期:2002-02-01 00:00:00

  • Expression and Purification of Recombinant Mouse Interleukin-4 and -6 from Transgenic Rice Seeds.

    abstract::Transgenic rice seed can be utilized as a bioreactor to produce high-value recombinant proteins. Mouse interleukin 4 (mIL-4) and mIL-6 were specifically expressed as secretory proteins in rice endosperm by ligating the N-terminal glutelin B-1 (GluB-1) signal peptide and the C-terminal KDEL endoplasmic reticulum retent...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-016-9920-7

    authors: Fujiwara Y,Yang L,Takaiwa F,Sekikawa K

    更新日期:2016-04-01 00:00:00

  • Online resources for SNP analysis: a review and route map.

    abstract::The major online single nucleotide polymorphism (SNP) databases freely available as research tools for genetic analysis are explained, reviewed, and compared. An outline is given of the search strategies that can be used with the most extensive current SNP databases: National Centre for Biotechnology Information (NCBI...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章,评审

    doi:10.1385/mb:35:1:65

    authors: Phillips C

    更新日期:2007-01-01 00:00:00

  • Genetic relationships among wild and cultivated accessions of curry leaf plant (Murraya koenigii (L.) Spreng.), as revealed by DNA fingerprinting methods.

    abstract::Murraya koenigii (L.) Spreng. (Rutaceae), is an aromatic plant and much valued for its flavor, nutritive and medicinal properties. In this study, three DNA fingerprinting methods viz., random amplification of polymorphic DNA (RAPD), directed amplification of minisatellite DNA (DAMD), and inter-simple sequence repeat (...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-012-9500-4

    authors: Verma S,Rana TS

    更新日期:2013-02-01 00:00:00

  • Comparison of Lentiviral Packaging Mixes and Producer Cell Lines for RNAi Applications.

    abstract::Lentiviral transduction is a highly efficient DNA delivery method for RNA interference applications. However, obtaining high lentiviral titers of shRNA and miRNA encoding vectors is challenging, since shRNA and miRNA cassettes have been shown to reduce lentiviral titers. In this study, we compare four commercially ava...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-015-9843-8

    authors: Albrecht C,Hosiner S,Tichy B,Aldrian S,Hajdu S,Nürnberger S

    更新日期:2015-06-01 00:00:00

  • The coupling of differential display and AFLP approaches for nonradioactive mRNA fingerprinting.

    abstract::We have modified the differential display of 3'-end restriction fragments of cDNA technique by combining it with the amplified fragment length polymorphism (AFLP) approach and silver staining. Modified oligo-dT primers were used for a reverse transcription step. ds cDNA was digested with the Mse I restriction enzyme a...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:12:2:137

    authors: Ivashuta S,Imai R,Uchiyama K,Gau M

    更新日期:1999-09-01 00:00:00

  • Lipofectamine RNAiMAX: an efficient siRNA transfection reagent in human embryonic stem cells.

    abstract::RNA interference methodology suppresses specific gene expression, thus mimicking loss-of-function mutation and enabling in vitro and in vivo gene function analysis. Lipofectamine RNAiMAX, a new transfection reagent, has been confirmed high efficiency in delivering small interfering RNA (siRNA) into mesenchymal stem ce...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-008-9043-x

    authors: Zhao M,Yang H,Jiang X,Zhou W,Zhu B,Zeng Y,Yao K,Ren C

    更新日期:2008-09-01 00:00:00

  • Responsiveness and Adaptation to Salt Stress of the REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) Gene are Controlled by its Promoter.

    abstract::The REDOX-RESPONSIVE TRANSCRIPTION FACTOR 1 (RRTF1) gene encodes a member of the ERF/AP2 transcription factor family involved in redox homeostasis. The RRTF1 gene shows tissue-specific responsiveness to various abiotic stress treatments including a response to salt stress in roots. An interesting feature of this respo...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-019-00155-9

    authors: Soliman ERS,Meyer P

    更新日期:2019-04-01 00:00:00

  • Rapid and effective method of RNA isolation from green microalga Ankistrodesmus convolutus.

    abstract::The rapid and effective method for the isolation of RNA from green microalga Ankistrodesmus convolutus based on homogenization in a simple CTAB buffer and selective precipitation of RNA with lithium chloride is developed. This procedure avoids the use of toxic chaotropic agents and phenol while high concentration of d...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-009-9182-8

    authors: Thanh T,Omar H,Abdullah MP,Chi VT,Noroozi M,Ky H,Napis S

    更新日期:2009-10-01 00:00:00

  • Inter- and intra-varietal genetic variability in Malvasia cultivars.

    abstract::The DNA molecular analyses together with ampelography, ampelometry, and biochemistry are essential for grapevine identification and investigation of genetic differences among the Vitis vinifera L. cultivars and clones. Ten Malvasia cultivars (i.e., Istrian Malvasia; M. delle Lipari; M. bianca di Candia; M. di Candia A...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-011-9423-5

    authors: Meneghetti S,Poljuha D,Frare E,Costacurta A,Morreale G,Bavaresco L,Calò A

    更新日期:2012-03-01 00:00:00

  • Characterization of Viral Genome Encapsidated in Adeno-associated Recombinant Vectors Produced in Yeast Saccharomyces cerevisiae.

    abstract::Adeno-associated virus (AAV) is a small, non-enveloped virus used as vector in gene therapy, mainly produced in human cells and in baculovirus systems. Intense studies on these platforms led to the production of vectors with titers between 103 and 105 viral genomes (vg) per cells. In spite of this, vector yields need ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-020-00294-4

    authors: Galli A,Iaia I,Milella MS,Cipriani F,Della Latta V,Giacca M,Zentilin L,Cervelli T

    更新日期:2021-02-01 00:00:00

  • Use of synthetic genes for cloning, production and functional expression of the bacteriocins enterocin A and bacteriocin E 50-52 by Pichia pastoris and Kluyveromyces lactis.

    abstract::The use of synthetic genes may constitute a successful approach for the heterologous production and functional expression of bacterial antimicrobial peptides (bacteriocins) by recombinant yeasts. In this work, synthetic genes with adapted codon usage designed from the mature amino acid sequence of the bacteriocin ente...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1007/s12033-014-9731-7

    authors: Jiménez JJ,Borrero J,Gútiez L,Arbulu S,Herranz C,Cintas LM,Hernández PE

    更新日期:2014-06-01 00:00:00

  • Preparation of peptide-loaded dendritic cells for cancer immunotherapy.

    abstract::Dendritic cell-based vaccines are being evaluated in clinical trials to determine their ability to activate clinically relevant tumor antigen-specific immune responses. Although some groups isolate dendritic cells from peripheral blood, most have found it more efficient to generate large numbers from peripheral blood ...

    journal_title:Molecular biotechnology

    pub_type: 杂志文章

    doi:10.1385/MB:25:1:95

    authors: Morse MA,Clay T,Colling K,Lyerly HK

    更新日期:2003-09-01 00:00:00