Identification of the gene for β-fructofuranosidase from Ceratocystis moniliformis CMW 10134 and characterization of the enzyme expressed in Saccharomyces cerevisiae.

Abstract:

BACKGROUND:β-Fructofuranosidases (or invertases) catalyse the commercially-important biotransformation of sucrose into short-chain fructooligosaccharides with wide-scale application as a prebiotic in the functional foods and pharmaceutical industries. RESULTS:We identified a β-fructofuranosidase gene (CmINV) from a Ceratocystis moniliformis genome sequence using protein homology and phylogenetic analysis. The predicted 615 amino acid protein, CmINV, grouped with an existing clade within the glycoside hydrolase (GH) family 32 and showed typical conserved motifs of this enzyme family. Heterologous expression of the CmINV gene in Saccharomyces cerevisiae BY4742∆suc2 provided further evidence that CmINV indeed functions as a β-fructofuranosidase. Firstly, expression of the CmINV gene complemented the inability of the ∆suc2 deletion mutant strain of S. cerevisiae to grow on sucrose as sole carbohydrate source. Secondly, the recombinant protein was capable of producing short-chain fructooligosaccharides (scFOS) when incubated in the presence of 10% sucrose. Purified deglycosylated CmINV protein showed a molecular weight of ca. 66 kDa and a Km and Vmax on sucrose of 7.50 mM and 986 μmol/min/mg protein, respectively. Its optimal pH and temperature conditions were determined to be 6.0 and 62.5°C, respectively. The addition of 50 mM LiCl led to a 186% increase in CmINV activity. Another striking feature was the relatively high volumetric production of this protein in S. cerevisiae as one mL of supernatant was calculated to contain 197 ± 6 International Units of enzyme. CONCLUSION:The properties of the CmINV enzyme make it an attractive alternative to other invertases being used in industry.

journal_name

BMC Biotechnol

journal_title

BMC biotechnology

authors

van Wyk N,Trollope KM,Steenkamp ET,Wingfield BD,Volschenk H

doi

10.1186/1472-6750-13-100

subject

Has Abstract

pub_date

2013-11-14 00:00:00

pages

100

issn

1472-6750

pii

1472-6750-13-100

journal_volume

13

pub_type

杂志文章
  • Construction and characterization of a high-quality cDNA library of Cymbidium faberi suitable for yeast one- and two-hybrid assays.

    abstract:BACKGROUND:Cymbidium faberi is one of the oldest cultivars of oriental orchids, with an elegant flower fragrance. In order to investigate the molecular mechanism and the functions of related proteins in the methyl jasmonate (MeJA) signaling pathway, one of the main components of flower fragrance in C. faberi, yeast one...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-020-0599-2

    authors: Xu Y,Zhou J,Liu Q,Li K,Zhou Y

    更新日期:2020-01-16 00:00:00

  • Ligation-anchored PCR unveils immune repertoire of TCR-beta from whole blood.

    abstract:BACKGROUND:As one of the genetic mechanisms for adaptive immunity, V(D)J recombination generates an enormous repertoire of T-cell receptors (TCRs). With the development of high-throughput sequencing techniques, systematic exploration of V(D)J recombination becomes possible. Multiplex PCR has been previously developed t...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0153-9

    authors: Gao F,Wang K

    更新日期:2015-05-28 00:00:00

  • Genetic and spectrally distinct in vivo imaging: embryonic stem cells and mice with widespread expression of a monomeric red fluorescent protein.

    abstract:BACKGROUND:DsRed the red fluorescent protein (RFP) isolated from Discosoma sp. coral holds much promise as a genetically and spectrally distinct alternative to green fluorescent protein (GFP) for application in mice. Widespread use of DsRed has been hampered by several issues resulting in the inability to establish and...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-5-20

    authors: Long JZ,Lackan CS,Hadjantonakis AK

    更新日期:2005-07-04 00:00:00

  • Mimotopes selected with a neutralizing antibody against urease B from Helicobacter pylori induce enzyme inhibitory antibodies in mice upon vaccination.

    abstract:BACKGROUND:Urease B is an important virulence factor that is required for Helicobacter pylori to colonise the gastric mucosa. Mouse monoclonal antibodies (mAbs) that inhibit urease B enzymatic activity will be useful as vaccines for the prevention and treatment of H. pylori infection. Here, we produced murine mAbs agai...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-10-84

    authors: Li Y,Ning Y,Wang Y,Peng D,Jiang Y,Zhang L,Long M,Luo J,Li M

    更新日期:2010-11-30 00:00:00

  • Identification of residues important for the activity of aldehyde-deformylating oxygenase through investigation into the structure-activity relationship.

    abstract:BACKGROUND:Aldehyde-deformylating oxygenase (ADO) is a key enzyme involved in the biosynthetic pathway of fatty alk(a/e)nes in cyanobacteria. However, cADO (cyanobacterial ADO) showed extreme low activity with the k cat value below 1 min-1, which would limit its application in biofuel production. To identify the activi...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-017-0351-8

    authors: Wang Q,Bao L,Jia C,Li M,Li JJ,Lu X

    更新日期:2017-03-16 00:00:00

  • An adenovirus prime/plasmid boost strategy for induction of equipotent immune responses to two dengue virus serotypes.

    abstract:BACKGROUND:Dengue is a public health problem of global significance for which there is neither an effective antiviral therapy nor a preventive vaccine. It is a mosquito-borne viral disease, caused by dengue (DEN) viruses, which are members of the Flaviviridae family. There are four closely related serotypes, DEN-1, DEN...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-7-10

    authors: Khanam S,Rajendra P,Khanna N,Swaminathan S

    更新日期:2007-02-15 00:00:00

  • Construction of sized eukaryotic cDNA libraries using low input of total environmental metatranscriptomic RNA.

    abstract:BACKGROUND:Construction of high quality cDNA libraries from the usually low amounts of eukaryotic mRNA extracted from environmental samples is essential in functional metatranscriptomics for the selection of functional, full-length genes encoding proteins of interest. Many of the inserts in libraries constructed by sta...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-14-80

    authors: Yadav RK,Barbi F,Ziller A,Luis P,Marmeisse R,Reddy MS,Fraissinet-Tachet L

    更新日期:2014-09-03 00:00:00

  • Susceptible cell lines for the production of porcine reproductive and respiratory syndrome virus by stable transfection of sialoadhesin and CD163.

    abstract:BACKGROUND:Porcine reproductive and respiratory syndrome virus (PRRSV) causes major economic losses in the pig industry worldwide. In vivo, the virus infects a subpopulation of tissue macrophages. In vitro, PRRSV only replicates in primary pig macrophages and African green monkey kidney derived cells, such as Marc-145....

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-10-48

    authors: Delrue I,Van Gorp H,Van Doorsselaere J,Delputte PL,Nauwynck HJ

    更新日期:2010-06-29 00:00:00

  • Setting up a platform for plant-based influenza virus vaccine production in South Africa.

    abstract:BACKGROUND:During a global influenza pandemic, the vaccine requirements of developing countries can surpass their supply capabilities, if these exist at all, compelling them to rely on developed countries for stocks that may not be available in time. There is thus a need for developing countries in general to produce t...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-12-14

    authors: Mortimer E,Maclean JM,Mbewana S,Buys A,Williamson AL,Hitzeroth II,Rybicki EP

    更新日期:2012-04-26 00:00:00

  • CdSe/ZnS quantum dots trigger DNA repair and antioxidant enzyme systems in Medicago sativa cells in suspension culture.

    abstract:BACKGROUND:Nanoparticles appear to be promising devices for application in the agriculture and food industries, but information regarding the response of plants to contact with nano-devices is scarce. Toxic effects may be imposed depending on the type and concentration of nanoparticle as well as time of exposure. A num...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-13-111

    authors: Santos AR,Miguel AS,Macovei A,Maycock C,Balestrazzi A,Oliva A,Fevereiro P

    更新日期:2013-12-20 00:00:00

  • Production of recombinant VP1-derived virus-like particles from novel human polyomaviruses in yeast.

    abstract:BACKGROUND:Eleven new human polyomaviruses (HPyVs) have been identified in the last decade. Serological studies show that these novel HPyVs sub-clinically infect humans at an early age. The routes of infection, entry pathways, and cell tropism of new HPyVs remain unknown. VP1 proteins of polyomaviruses can assembly int...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0187-z

    authors: Norkiene M,Stonyte J,Ziogiene D,Mazeike E,Sasnauskas K,Gedvilaite A

    更新日期:2015-08-04 00:00:00

  • Rapid high-yield expression of a candidate influenza vaccine based on the ectodomain of M2 protein linked to flagellin in plants using viral vectors.

    abstract:BACKGROUND:The extracellular domain of matrix protein 2 (M2e) of influenza A virus is a promising target for the development of a universal vaccine against influenza because M2e sequences are highly conserved among human influenza A strains. However, native M2e is poorly immunogenic, but its immunogenicity can be incre...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0164-6

    authors: Mardanova ES,Kotlyarov RY,Kuprianov VV,Stepanova LA,Tsybalova LM,Lomonosoff GP,Ravin NV

    更新日期:2015-05-29 00:00:00

  • Modification of the loops in the ligand-binding site turns avidin into a steroid-binding protein.

    abstract:BACKGROUND:Engineered proteins, with non-immunoglobulin scaffolds, have become an important alternative to antibodies in many biotechnical and therapeutic applications. When compared to antibodies, tailored proteins may provide advantageous properties such as a smaller size or a more stable structure. RESULTS:Avidin i...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-11-64

    authors: Riihimäki TA,Hiltunen S,Rangl M,Nordlund HR,Määttä JA,Ebner A,Hinterdorfer P,Kulomaa MS,Takkinen K,Hytönen VP

    更新日期:2011-06-09 00:00:00

  • A miniaturized sandwich immunoassay platform for the detection of protein-protein interactions.

    abstract:BACKGROUND:Analysis of protein-protein interactions (PPIs) is a valuable approach for the characterization of huge networks of protein complexes or proteins of unknown function. Co-immunoprecipitation (coIP) using affinity resins coupled to protein A/G is the most widely used method for PPI detection. However, this tra...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-10-78

    authors: Liu Q,Chen Q,Wang J,Zhang Y,Zhou Y,Lin C,He W,He F,Xu D

    更新日期:2010-10-28 00:00:00

  • Production of soluble mammalian proteins in Escherichia coli: identification of protein features that correlate with successful expression.

    abstract:BACKGROUND:In the search for generic expression strategies for mammalian protein families several bacterial expression vectors were examined for their ability to promote high yields of soluble protein. Proteins studied included cell surface receptors (Ephrins and Eph receptors, CD44), kinases (EGFR-cytoplasmic domain, ...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-4-32

    authors: Dyson MR,Shadbolt SP,Vincent KJ,Perera RL,McCafferty J

    更新日期:2004-12-14 00:00:00

  • Specific and sensitive quantitative RT-PCR of miRNAs with DNA primers.

    abstract:BACKGROUND:MicroRNAs are important regulators of gene expression at the post-transcriptional level and play an important role in many biological processes. Due to the important biological role it is of great interest to quantitatively determine their expression level in different biological settings. RESULTS:We descri...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-11-70

    authors: Balcells I,Cirera S,Busk PK

    更新日期:2011-06-25 00:00:00

  • Dynamic in vivo imaging and cell tracking using a histone fluorescent protein fusion in mice.

    abstract:BACKGROUND:Advances in optical imaging modalities and the continued evolution of genetically-encoded fluorescent proteins are coming together to facilitate the study of cell behavior at high resolution in living organisms. As a result, imaging using autofluorescent protein reporters is gaining popularity in mouse trans...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-4-33

    authors: Hadjantonakis AK,Papaioannou VE

    更新日期:2004-12-24 00:00:00

  • Characterization of mutations and loss of heterozygosity of p53 and K-ras2 in pancreatic cancer cell lines by immobilized polymerase chain reaction.

    abstract:BACKGROUND:The identification of known mutations in a cell population is important for clinical applications and basic cancer research. In this work an immobilized form of the polymerase chain reaction, referred to as polony technology, was used to detect mutations as well as gene deletions, resulting in loss of hetero...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-3-11

    authors: Butz J,Wickstrom E,Edwards J

    更新日期:2003-07-23 00:00:00

  • The relationship between mTOR signalling pathway and recombinant antibody productivity in CHO cell lines.

    abstract:BACKGROUND:High recombinant protein productivity in mammalian cell lines is often associated with phenotypic changes in protein content, energy metabolism, and cell growth, but the key determinants that regulate productivity are still not clearly understood. The mammalian target of rapamycin (mTOR) signalling pathway h...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-14-15

    authors: Edros R,McDonnell S,Al-Rubeai M

    更新日期:2014-02-17 00:00:00

  • A novel psychrophilic alkaline phosphatase from the metagenome of tidal flat sediments.

    abstract:BACKGROUND:Alkaline phosphatase (AP) catalyzes the hydrolytic cleavage of phosphate monoesters under alkaline conditions and plays important roles in microbial ecology and molecular biology applications. Here, we report on the first isolation and biochemical characterization of a thermolabile AP from a metagenome. RES...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0115-2

    authors: Lee DH,Choi SL,Rha E,Kim SJ,Yeom SJ,Moon JH,Lee SG

    更新日期:2015-01-31 00:00:00

  • Secretion of Fc-amidated peptide fusion proteins by Chinese hamster ovary cells.

    abstract:BACKGROUND:The therapeutic use of α-amidated peptides (e.g. calcitonin, glucagon-like peptide) has increased dramatically, but there are major impediments to wider use of such peptides. Larger peptides are expensive to synthesize, and short plasma half-lives frequently limit the clinical circumstances in which the pept...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0173-5

    authors: Carlson KR,Pomerantz SC,Li J,Vafa O,Naso M,Strohl W,Mains RE,Eipper BA

    更新日期:2015-06-27 00:00:00

  • Dye decolorization and detoxification potential of Ca-alginate beads immobilized manganese peroxidase.

    abstract:BACKGROUND:In view of compliance with increasingly stringent environmental legislation, an eco-friendly treatment technology of industrial dyes and effluents is a major environmental challenge in the color industry. In present study, a promising and eco-friendly entrapment approach was adopted to immobilize purified ma...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-015-0227-8

    authors: Bilal M,Asgher M

    更新日期:2015-12-10 00:00:00

  • Microporation is a valuable transfection method for efficient gene delivery into human umbilical cord blood-derived mesenchymal stem cells.

    abstract:BACKGROUND:Mesenchymal stem cells (MSCs) are an attractive source of adult stem cells for therapeutic application in clinical study. Genetic modification of MSCs with beneficial genes makes them more effective for therapeutic use. However, it is difficult to transduce genes into MSCs by common transfection methods, esp...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-10-38

    authors: Lim JY,Park SH,Jeong CH,Oh JH,Kim SM,Ryu CH,Park SA,Ahn JG,Oh W,Jeun SS,Chang JW

    更新日期:2010-05-13 00:00:00

  • Efficient CPP-mediated Cre protein delivery to developing and adult CNS tissues.

    abstract:BACKGROUND:Understanding and manipulating gene function in physiological conditions is a major objective for both fundamental and applied research. In contrast to other experimental settings, which use either purely genetic or gene delivery (viral or non-viral) strategies, we report here a strategy based on direct prot...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-9-40

    authors: Gitton Y,Tibaldi L,Dupont E,Levi G,Joliot A

    更新日期:2009-04-24 00:00:00

  • A novel expression system of domain I of human beta2 glycoprotein I in Escherichia coli.

    abstract:BACKGROUND:The antiphospholipid syndrome (APS), characterised by recurrent miscarriage and thrombosis, is a significant cause of morbidity and mortality. Domain I (DI) of human beta 2 glycoprotein I (beta2GPI) is thought to contain crucial antibody binding epitopes for antiphospholipid antibodies (aPL), which are criti...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-6-8

    authors: Ioannou Y,Giles I,Lambrianides A,Richardson C,Pearl LH,Latchman DS,Isenberg DA,Rahman A

    更新日期:2006-02-10 00:00:00

  • Thin films of Type 1 collagen for cell by cell analysis of morphology and tenascin-C promoter activity.

    abstract:BACKGROUND:The use of highly reproducible and spatiallyhomogeneous thin film matrices permits automated microscopy and quantitative determination of the response of hundreds of cells in a population. Using thin films of extracellular matrix proteins, we have quantified, on a cell-by-cell basis, phenotypic parameters of...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-6-14

    authors: Langenbach KJ,Elliott JT,Tona A,McDaniel D,Plant AL

    更新日期:2006-03-06 00:00:00

  • Switchgrass (Panicum virgatum L.) polyubiquitin gene (PvUbi1 and PvUbi2) promoters for use in plant transformation.

    abstract:BACKGROUND:The ubiquitin protein is present in all eukaryotic cells and promoters from ubiquitin genes are good candidates to regulate the constitutive expression of transgenes in plants. Therefore, two switchgrass (Panicum virgatum L.) ubiquitin genes (PvUbi1 and PvUbi2) were cloned and characterized. Reporter constru...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-11-74

    authors: Mann DG,King ZR,Liu W,Joyce BL,Percifield RJ,Hawkins JS,LaFayette PR,Artelt BJ,Burris JN,Mazarei M,Bennetzen JL,Parrott WA,Stewart CN Jr

    更新日期:2011-07-11 00:00:00

  • Pattern analysis approach reveals restriction enzyme cutting abnormalities and other cDNA library construction artifacts using raw EST data.

    abstract:BACKGROUND:Expressed Sequence Tag (EST) sequences are widely used in applications such as genome annotation, gene discovery and gene expression studies. However, some of GenBank dbEST sequences have proven to be "unclean". Identification of cDNA termini/ends and their structures in raw ESTs not only facilitates data qu...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-12-16

    authors: Zhou S,Ji G,Liu X,Li P,Moler J,Karro JE,Liang C

    更新日期:2012-05-03 00:00:00

  • Heterogeneous oxygen availability affects the titer and topology but not the fidelity of plasmid DNA produced by Escherichia coli.

    abstract:BACKGROUND:Dissolved oxygen tension (DOT) is hardly constant and homogenously distributed in a bioreactor, which can have a negative impact in the metabolism and product synthesis. However, the effects of DOT on plasmid DNA (pDNA) production and quality have not been thoroughly investigated. In the present study, the e...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/s12896-017-0378-x

    authors: Jaén KE,Sigala JC,Olivares-Hernández R,Niehaus K,Lara AR

    更新日期:2017-07-04 00:00:00

  • Generation and characterization of a human single-chain fragment variable (scFv) antibody against cytosine deaminase from yeast.

    abstract:BACKGROUND:The ability of cytosine deaminase (CD) to convert the antifungal agent 5-fluorocytosine (5-FC) into one of the most potent and largely used anticancer compound such as 5-fluorouracil (5-FU) raised considerable interest in this enzyme to model gene or antibody - directed enzyme-prodrug therapy (GDEPT/ADEPT) a...

    journal_title:BMC biotechnology

    pub_type: 杂志文章

    doi:10.1186/1472-6750-8-68

    authors: Mallano A,Zamboni S,Carpinelli G,Santoro F,Flego M,Ascione A,Gellini M,Tombesi M,Podo F,Cianfriglia M

    更新日期:2008-09-10 00:00:00