Rational design of thermostable lactate oxidase by analyzing quaternary structure and prevention of deamidation.


:Our current knowledge of protein unfolding is overwhelmingly related to reversible denaturation. However, to engineer thermostable enzymes for industrial applications and medical diagnostics, it is necessary to consider irreversible denaturation processes and/or the entire quaternary structure. In this study we have used lactate oxidase (LOD), which is employed in lactic acid sensors, as a model example to design thermostable variants by rational design. Twelve mutant proteins were tested and one of them displayed a markedly greater thermostability than all the mutants we had previously obtained by random mutagenesis. This mutant was designed so as to strengthen the interaction between the subunits and stabilize the quaternary structure. Since LOD is difficult to crystallize, its three-dimensional structure remains unknown. This study shows that it is possible to carry out rational design to improve thermostability using a computer-aided quaternary structure model based on the known tertiary structure of a related protein. Critical factors required for increasing the thermal stability of proteins by rational design, where the 3-D structure is not available, are discussed.


Biotechnol Lett


Biotechnology letters


Kaneko H,Minagawa H,Shimada J





Has Abstract


2005-11-01 00:00:00












  • Isolation and characterization of the citrinin biosynthetic gene cluster from Monascus aurantiacus.

    abstract::Monascus aurantiacus produces high amounts of citrinin which is a mycotoxin with nephrotoxic activity. Six putative citrinin biosynthesis genes have been discovered in M. purpureus and at least 10 genes are responsible for its biosynthesis. However, the sequence of citrinin pathway gene cluster in M. aurantiacus has n...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Li YP,Xu Y,Huang ZB

    更新日期:2012-01-01 00:00:00

  • Biofilm productivity and concomitant cell autolysis in a membrane bioreactor.

    abstract::Phanerochaete chrysoporium morphology and manganese peroxidase (MnP) productivity was characterised in a scalable, modularised 1145 cm(3) membrane gradostat reactor in response to switching between an enhanced production medium and a nutrient limited feed (50% C and N reduction). Irrespective of the feed composition u...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Govender S

    更新日期:2011-02-01 00:00:00

  • Removing chiral contamination of lactate solutions by selective metabolism of the D-enantiomer.

    abstract:OBJECTIVE:A bio-based process is appealing for purification of L-lactic acid, the major enantiomer of polylactic acid syrup, generated by thermochemical processes at the end of life of PLA-based plastics, from its chiral impurity, D-lactic acid, before reuse. RESULTS:Polylactic acid (PLA), a renewable alternative to p...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Chauliac D,Pullammanappallil PC,Ingram LO,Shanmugam KT

    更新日期:2015-12-01 00:00:00

  • Genome shuffling improves production of the low-temperature alkalophilic lipase by Acinetobacter johnsonii.

    abstract::The production of a low-temperature alkalophilic lipase from Acinetobacter johnsonii was improved using genome shuffling. The starting populations, obtained by UV irradiation and diethyl sulfate mutagenesis, were subjected to recursive protoplast fusion. The optimal conditions for protoplast formation and regeneration...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Wang H,Zhang J,Wang X,Qi W,Dai Y

    更新日期:2012-01-01 00:00:00

  • Recent progress and new challenges in metagenomics for biotechnology.

    abstract::A brief historical perspective on metagenomics is given followed by a discussion of the rapid progress in this field largely defined by transition to the next generation sequencing technologies. Problems and challenges connected to this transition are also addressed. The review focuses on recent literature describing ...

    journal_title:Biotechnology letters

    pub_type: 历史文章,杂志文章,评审


    authors: Chistoserdova L

    更新日期:2010-10-01 00:00:00

  • Improved recovery of a fusion protein containing the antigenic domain 1 of the human cytomegalovirus glycoprotein B.

    abstract::Heterologous expression in Escherichia coli often leads to production of the expressed proteins as insoluble and inactive inclusion bodies. The general strategy for protein recovery includes isolation and washing of inclusion bodies, solubilization of aggregated protein and refolding of solubilized protein. The proces...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Ferreira S,Sousa F,Queiroz JA,Domingues FC

    更新日期:2005-08-01 00:00:00

  • Prevention of osteoporosis in ovariectomized mice by electroporational gene transfer of parathyroid hormone.

    abstract::Non-viral vector, pCMV-Pth, with full-length human parathyroid hormone (prepro-hPTH) cDNA was constructed and delivered into the ovariectomized mouse quadriceps to explore the effects of electroporational gene transfer of PTH on the bone. The expression of hPTH in the transfected mice was detected by RT-PCR and radioi...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Liu B,Tang J,Ji J,Gu J

    更新日期:2003-12-01 00:00:00

  • Novel Escherichia coli hybrids with enhanced butanol tolerance.

    abstract::Hybrids between Escherichia coli and Lactobacillus brevis were generated via protoplast fusion. Growth kinetics of five hybrid strains and E. coli were used to evaluate the butanol tolerance of the novel strains under different conditions. The hybrid strains tolerated up to 2% (v/v) butanol compared to the 1% (v/v) ma...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Winkler J,Rehmann M,Kao KC

    更新日期:2010-07-01 00:00:00

  • Synthesis, expression and purification of a type of chlorotoxin-like peptide from the scorpion, Buthus martensii Karsch, and its acute toxicity analysis.

    abstract::A gene, rBmK Cta, encoding a chlorotoxin-like peptide from the scorpion, Buthus martensii Karsch, was synthesized according to the sequence optimized for codon usage in Escherichia coli and was expressed in E. coli BL21 (DE3) using a pExSecI expression system in which the IgG-binding domain-ZZ of protein A is fused to...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Fu YJ,Yin LT,Wang W,Chai BF,Liang AH

    更新日期:2005-10-01 00:00:00

  • Expression of the human cancer/testis antigen NY-SAR-35 is activated by CpG island hypomethylation.

    abstract::The novel cancer/testis antigen gene, NY-SAR-35, is expressed exclusively in normal testis and in various histological types of tumor. However, the NY-SAR-35 gene expression is observed to be aberrant in several cancer cell lines and tissues. The analysis of methylation status of the NY-SAR-35 gene promoter in various...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Park JH,Song MH,Lee CH,Lee MK,Park YM,Old L,Lee SY

    更新日期:2011-06-01 00:00:00

  • Stability of high cell density brewery fermentations during serial repitching.

    abstract::The volumetric productivity of the beer fermentation process can be increased by using a higher pitching rate (i.e. higher inoculum size). However, the decreased yeast net growth observed in these high cell density brewery fermentations can adversely affect the physiological stability throughout subsequent yeast gener...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Verbelen PJ,Dekoninck TM,Van Mulders SE,Saerens SM,Delvaux F,Delvaux FR

    更新日期:2009-11-01 00:00:00

  • Oral administration of recombinant human granulocyte-macrophage colony stimulating factor expressed in rice endosperm can increase leukocytes in mice.

    abstract::Human granulocyte-macrophage colony stimulating factor (hGM-CSF) is used clinically to treat leucopenia typically caused by cancer chemotherapy or radiotherapy. This study used multiple strategies to obtain very high expression levels of OsrhGM-CSF (14 microg/seed) in rice endosperm. Electron micrographs of immunogold...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Ning T,Xie T,Qiu Q,Yang W,Zhou S,Zhou L,Zheng C,Zhu Y,Yang D

    更新日期:2008-09-01 00:00:00

  • Expandase-like activity mediated cell-free conversion of ampicillin to cephalexin by Streptomyces sp. DRS I.

    abstract::Cell-free extracts of Streptomyces sp. DRS I converted ampicillin to cephalexin, presumably due to the activity of the enzyme, expandase. The extract was fractionated and characterized by colorimetric and chromatographic measurements coupled with disc-agar diffusion bioassay against an ampicillin-resistant, cephalexin...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Thakur D,Roy MK,Bora TC

    更新日期:2009-07-01 00:00:00

  • Improvement of glutathione production by a metabolically engineered Yarrowia lipolytica strain using a small-scale optimization approach.

    abstract:OBJECTIVE:In this study, we aimed to maximize glutathione (GSH) production by a metabolically engineered Yarrowia lipolytica strain using a small-scale optimization approach. RESULTS:A three levels four factorial Box-Behnken Design was used to assess the effect of pH, inulin extract, yeast extract and ammonium sulfate...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Do DTH,Fickers P,Ben Tahar I

    更新日期:2021-02-01 00:00:00

  • Biosurfactant production by Serratia marcescens SS-1 and its isogenic strain SMdeltaR defective in SpnR, a quorum-sensing LuxR family protein.

    abstract::Serratia marcescens SS-1 and its SpnR-defective isogenic mutant, SMdeltaR, produced an extracellular surfactant able to decrease surface tension of water from 72 to 37 dyne cm(-1) (SMdeltaR strain) and to 45 dyne cm(-1) (SS-1 strain). The biosurfactant also emulsified kerosene and diesel with a maximum emulsion index ...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Wei YH,Lai HC,Chen SY,Yeh MS,Chang JS

    更新日期:2004-05-01 00:00:00

  • On-column refolding and purification of recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) expressed as inclusion body in Escherichia coli.

    abstract::Recombinant human interleukin-1 receptor antagonist (rHuIL-1ra) was produced in E. coli as an inclusion body. rHuIL-1ra was purified to Over 98% purity by anion exchange chromatography after on-column refolding. The optimized processes produced more than 2 g pure refolded rHuIL-1ra per 1 l culture, corresponding to a ...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Tan H,Dan G,Gong H,Cao L

    更新日期:2005-08-01 00:00:00

  • A new irreversible enzyme-aided esterification method in organic solvents.

    abstract::A new irreversible esterification method for carboxylic acids catalyzed by a lipase from Candida antarctica (Novozyme 435) in organic solvents has been developed. The water produced during the process is chemically destroyed by a corresponding ester of acetoacetate, which acts as a sacrificial substrate in this reacti...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Jeromin GE,Zoor A

    更新日期:2008-05-01 00:00:00

  • Enhancement of phase separation by the addition of de-emulsifiers to three-phase (diesel oil/biocatalyst/aqueous phase) emulsion in diesel biodesulfurization.

    abstract::Ethanol, added as a de-emulsifier to separate oil and biocatalyst (or bacterial cells) from a three-phase (oil/biocatalyst/aqueous phase) emulsion, formed in diesel biodesulfurization employing Gordonia nitida, improved oil recovery by centrifugation from about 50% in its absence to almost 100% at 3% (v/v). The biocat...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Choi OK,Choi KS,Ryu HW,Chang YK

    更新日期:2003-01-01 00:00:00

  • Isolation of an Acremonium sp. from a screening of 52 seawater fungal isolates and preliminary characterization of its growth conditions and nematicidal activity.

    abstract::Fifty-two fungal isolates obtained from seawater were screened for anti-nematode activity. One isolate, an Acremonium sp., exhibited the highest activity against the pinewood nematode, Bursaphelenchus xylophilus. When it was grown at pH 7.0 and 24-26 °C, nematicidal metabolites were produced. These were heat-stable, w...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Meng Q,Shi X,Meng F,Feng X,Sun J

    更新日期:2012-10-01 00:00:00

  • Lipid production from indigenous Greek microalgae: a possible biodiesel source.

    abstract:OBJECTIVE:Microalgae gained interest for potential use as biodiesel producers, since they synthesize and accumulate significant quantities of lipids. The aim of this work was to isolate indigenous microalgae strains from Greek habitats, study their physicochemical growth conditions and finally select the best ones with...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Savvides AL,Moisi K,Katsifas EA,Karagouni AD,Hatzinikolaou DG

    更新日期:2019-05-01 00:00:00

  • Biocatalytic characterization of a short-chain alcohol dehydrogenase with broad substrate specificity from thermophilic Carboxydothermus hydrogenoformans.

    abstract::The gene encoding a novel short-chain alcohol dehydrogenase in the thermophilic bacterium, Carboxydothermus hydrogenoformans, was identified and overexpressed in Escherichia coli. The enzyme was thermally stable and displayed the highest activity at 70 °C and pH 6.0. It preferred NAD(H) over NADP(H) as a cofactor and ...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Zhou S,Zhang SC,Lai DY,Zhang SL,Chen ZM

    更新日期:2013-03-01 00:00:00

  • Production of trans-10,cis-12-conjugated linoleic acid using permeabilized whole-cell biocatalyst of Yarrowia lipolytica.

    abstract:OBJECTIVE:To improve the production of trans-10,cis-12-conjugated linoleic acid (t10,c12-CLA) from linoleic acid in recombinant Yarrowia lipolytica. RESULTS:Cells of the yeast were permeabilized by freeze/thawing. The optimal conditions for t10,c12-CLA production by the permeabilized cells were at 28 °C, pH 7, 200 rpm...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Zhang B,Song Y,Chen H,Zhang H,Chen W

    更新日期:2016-11-01 00:00:00

  • Co-expression of chimeric chitinase and a polygalacturonase-inhibiting protein in transgenic canola (Brassica napus) confers enhanced resistance to Sclerotinia sclerotiorum.

    abstract:OBJECTIVES:Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum is one of the major fungal diseases of canola. To develop resistance against this fungal disease, the chit42 from Trichoderma atroviride with chitin-binding domain and polygalacturonase-inhibiting protein 2 (PG1P2) of Phaseolus vulgaris were co-ex...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Ziaei M,Motallebi M,Zamani MR,Panjeh NZ

    更新日期:2016-06-01 00:00:00

  • Immobilization of Pseudomonas cepacia lipase onto electrospun polyacrylonitrile fibers through physical adsorption and application to transesterification in nonaqueous solvent.

    abstract::The lipase of Pseudomonas cepacia was immobilized onto electrospun polyacrylonitrile (PAN) fibers and used for the conversion of (S)-glycidol with vinyl n-butyrate to glycidyl n-butyrate in isooctane. The rate of reaction with the adsorbed lipase was 23-fold higher than the initial material. After 10 recyclings, the i...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Sakai S,Liu Y,Yamaguchi T,Watanabe R,Kawabe M,Kawakami K

    更新日期:2010-08-01 00:00:00

  • Identification of key pathways and differentially expressed genes in bronchopulmonary dysplasia using bioinformatics analysis.

    abstract:OBJECTIVES:The objective of this study was to discover unknown differentially expressed genes (DEGs) associated with bronchopulmonary dysplasia (BPD), analyze their functions and enriched signaling pathways, and identify hub genes correlating with BPD incidence and evolvement. RESULTS:Of 1289 DEGs identified, 568 were...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Yan W,Jiang M,Zheng J

    更新日期:2020-12-01 00:00:00

  • Increase in synthesis of human monoclonal antibodies by transfected Sp2/0 myeloma mouse cell line under conditions of microgravity.

    abstract::Microgravity can influence cell growth and function. A transfected Sp2/0 myeloma cell line P3A2 producing a human IgG1 anti-TNFa monoclonal antibody was cultivated in static culture, spinner flasks and simulated microgravity using a rotating wall vessel bioreactor. Microgravity significantly decreased cell growth (fro...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Foster LJ,Catzel D,Atwa S,Zarka M,Mahler SM

    更新日期:2003-08-01 00:00:00

  • The effects of polymer phase ratio and feeding strategy on solid-liquid TPPBs for the production of L-phenylacetylcarbinol from benzaldehyde using Candida utilis.

    abstract:PURPOSE OF WORK:To increase the bioproduction of L-phenylacetylcarbinol (PAC), a precursor molecule in the synthesis of the decongestants ephedrine and pseudoephedrine and which suffers from substrate, product, and by-product inhibition, by ensuring that the delivery of the substrate, benzaldehyde, is maintained within...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Khan TR,Daugulis AJ

    更新日期:2011-01-01 00:00:00

  • Inhibition of berberine hydrochloride on Candida albicans biofilm formation.

    abstract:OBJECTIVE:This study aimed to investigate the inhibitory effect of berberine hydrochloride (BH) on Candida albicans (C.albicans) ATCC10231 biofilm formation. RESULTS:This paper found a positive correlation between the concentration of BH and its inhibitory effect on the cellular activity of early biofilms because we f...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Huang X,Zheng M,Yi Y,Patel A,Song Z,Li Y

    更新日期:2020-11-01 00:00:00

  • Effect of bacterial cell size on electricity generation in a single-compartmented microbial fuel cell.

    abstract::A single-compartmented microbial fuel cell composed of a graphite felt anode modified with Neutral Red (NR-anode) and a porous Fe(II)-carbon cathode (FeC-cathode) were compared for electricity generation from Microbacterium sp. and Pseudomonas sp. under identical conditions. Pseudomonas sp. was more than four times th...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Lee SW,Jeon BY,Park DH

    更新日期:2010-04-01 00:00:00

  • Expression of cellulase genes in Saccharomyces cerevisiae via δ-integration subject to auxotrophic markers.

    abstract::δ-Integration can improve the expression stability and increase the copy number of exogenous genes in Saccharomyces cerevisiae. Generally, δ-integration vectors employ auxotrophic markers, such as LEU2-d, HIS3, TRP1, and URA3, to screen transformants. In this study, two cellulase genes (β-glucosidase and endoglucanase...

    journal_title:Biotechnology letters

    pub_type: 杂志文章


    authors: Liu L,Liu C,Zou S,Yang H,Hong J,Ma Y,Zhang M

    更新日期:2013-08-01 00:00:00