Rapid purification of truncated Taq DNA polymerase Stoffel fragment by boiling lysis of bacterial expression cultures.

Abstract:

:A simplified and low-cost method, boiling lysis, was developed and used to purify the truncated DNA polymerase Stoffel fragment after overproduction of the enzyme in Escherichia coli. The method is based on the thermostable properties of the Stoffel fragment. Conditions such as the boiling time and number of cycles employed were identified as determinants with which to achieve a high yield and activity of this truncated Taq enzyme. Under established conditions, a period of 8 min consisting of two cycles of boiling and centrifuging was ideal for the purification of the Taq protein, followed by dibasic phosphate and ethanol extraction in a single type of storage buffer to remove contaminating nucleic acid. The whole purification process could be achieved within 1 or 2 h. A total yield of 80 mg of truncated Taq enzyme with an activity of 1.6x10(7) units was purified from l litre of bacterial culture. The purified Taq enzyme had a molecular mass of 61 kDa, a value consistent with the truncated DNA polymerase Stoffel fragment reported previously.

authors

Yang Z,Ding Y,Zhang Y,Liu F

doi

10.1042/BA20070114

subject

Has Abstract

pub_date

2008-06-01 00:00:00

pages

71-5

issue

Pt 2

eissn

0885-4513

issn

1470-8744

pii

BA20070114

journal_volume

50

pub_type

杂志文章
  • Inhibitor studies of isopentenyl pyrophosphate biosynthesis in suspension cultures of the yew Taxus chinensis var. mairei.

    abstract::Some metabolic inhibitors, mevastatin (MVS), chlorocholine chloride (CCC), sodium pyrophosphate (NaPP) and D,L-glyceraldehyde (DLG), were respectively added into the suspension cultures of the yew Taxus chinensis var. mairei at the late phase of cell growth to study isopentenyl pyrophosphate (IPP) biosynthesis. The co...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/ba20020061

    authors: Wang YD,Yuan YJ,Lu M,Wu JC,Jiang JL

    更新日期:2003-02-01 00:00:00

  • Renal oxidative stress and nitric oxide production in streptozotocin-induced diabetic nephropathy in rats: the possible modulatory effects of garlic (Allium sativum L.).

    abstract::The purpose of the present study was to investigate the effects of garlic (Allium sativum L.) on the diabetic nephropathy and oxidative stress induced by STZ (streptozotocin) in rats. Diabetes was induced in Male Sprague-Dawley rats by administering a single intraperitoneal injection of STZ (60 mg/kg of body weight). ...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20080086

    authors: Mariee AD,Abd-Allah GM,El-Yamany MF

    更新日期:2009-03-01 00:00:00

  • Anthrax toxin-mediated delivery of cholera toxin-A subunit into the cytosol of mammalian cells.

    abstract::The protective antigen (PA) component of anthrax toxin mediates delivery of either lethal factor (LF) or oedema factor into the cytosol of mammalian cells. The N-terminal domain of LF(1-254) (amino acids 1-254 of LF) binds to PA and, when fused to heterologous proteins, delivers such proteins into the cytosol. In the ...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/ba20000037

    authors: Sharma M,Khanna H,Arora N,Singh Y

    更新日期:2000-08-01 00:00:00

  • Staphylococcus bacteriophage tails with bactericidal properties: new findings.

    abstract::The development of lytic bacteriophages as therapeutic products is an attractive alternative to antibiotics. In this study, we evaluated the potential of phage tails for lysing Gram-positive bacteria. Phage P954, a well-characterized temperate staphylococcal phage, was found to adsorb to a large number of Staphylococc...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.1052

    authors: Biddappa AC,Sundarrajan S,Ramalinga AB,Sriram B,Padmanabhan S

    更新日期:2012-11-01 00:00:00

  • Production and properties of a raw-starch-degrading amylase from the thermophilic and alkaliphilic Bacillus sp. TS-23.

    abstract::The optimum temperature and initial medium pH for amylase production by Bacillus sp. TS-23 were 55 degrees C and 8.5 respectively. Maximum amylase activity was obtained in a medium containing peptone and soluble starch as nitrogen and carbon sources. Activity staining revealed that two amylases with molecular masses o...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:

    authors: Lin LL,Chyau CC,Hsu WH

    更新日期:1998-08-01 00:00:00

  • Effect of oxidative stress on the production of recombinant human interferon-gamma in Escherichia coli.

    abstract::To reduce the degradation pathway of rhIFN-gamma (recombinant human interferon-gamma), the susceptibility against oxidative stress during fermentation in Escherichia coli cells and in purification process were investigated. Fermentations of recombinant E. coli were performed at 5, 30 and 60% DO (dissolved oxygen) conc...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20030230

    authors: Mohammadian-Mosaabadi J,Naderi-Manesh H,Maghsoudi N,Khalilzadeh R,Shojaosadati SA,Ebrahimi M

    更新日期:2005-02-01 00:00:00

  • Overall volumetric oxygen transfer coefficient in an aerated bench-top stirred fermenter in aqueous dispersions of sodium alginate.

    abstract::The dynamic gassing-out method was used to assess the effect of stirrer speed, air flow rate and alginate concentration on the overall volumetric oxygen mass transfer coefficient (K(L)a) in a bench-top stirred fermenter equipped with two Rushton-type turbines. As the alginate concentration in the medium was increased ...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20030213

    authors: Parente E,Piraino P,Fidaleo M,Moresi M

    更新日期:2004-10-01 00:00:00

  • Bioconversion of lignin and its derivatives into polyhydroxyalkanoates: Challenges and opportunities.

    abstract::Renewable energy resources are considered to be promising for the development of a sustainable circular economy. Among various alternatives, the microbial route for various biofuels production is quite lucrative. Use of cellulose and lignocellulose for methane, H2 , organic acids, ethanol, and cellulase has been explo...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章,评审

    doi:10.1002/bab.1720

    authors: Kumar P,Maharjan A,Jun HB,Kim BS

    更新日期:2019-03-01 00:00:00

  • A kinetic study of bovine haemoglobin hydrolysis by pepsin immobilized on a functionalized alumina to prepare hydrolysates containing bioactive peptides.

    abstract::The hydrolysis kinetics of native and denatured haemoglobin, using pepsin immobilized on aluminium oxide, was studied in order to produce hydrolysates containing bioactive peptides. Pepsin was immobilized on acidic alumina and on 2-ethanolamine- O -phosphate (2-EAOP)-modified acidic alumina. Surface charge of the two ...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20030131

    authors: Ticu EL,Vercaigne-Marko D,Huma A,Artenie V,Toma O,Guillochon D

    更新日期:2004-04-01 00:00:00

  • Novozym 435-catalysed transesterification of crude soya bean oils for biodiesel production in a solvent-free medium.

    abstract::When crude soya-bean oil was used as the source for biodiesel production, the yield of methyl ester was significantly lower than that from refined soya-bean oil. The major difference between refined and crude soya-bean oils was found to be due to the contents of phospholipids, free acid and water, which have varied in...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20030142

    authors: Du W,Xu YY,Zeng J,Liu DH

    更新日期:2004-10-01 00:00:00

  • Self-assembling bilayer lipid membranes on solid support.

    abstract::Solid-supported bilayer lipid membranes (s-BLMs) that possess some properties similar to those of conventional BLMs can be self-assembled on a freshly cleaved metal wire by a two-step procedure: (i) The tip of a Teflon-coated platinum wire, while immersed in a lipid solution, is cut off with a scalpel; (ii) the new ti...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Tien HT,Salamon Z

    更新日期:1990-10-01 00:00:00

  • Towards molecular farming in the future: pichia pastoris-based production of single-chain antibody fragments.

    abstract::This review article focuses on the use of the methylotrophic yeast Pichia pastoris as a recombinant protein-expression system. P. pastoris is a useful system for the expression of milligram-to-gram quantities of a protein, which can be scaled up to fermentation to meet greater demands. Compared with mammalian cells, P...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章,评审

    doi:

    authors: Fischer R,Drossard J,Emans N,Commandeur U,Hellwig S

    更新日期:1999-10-01 00:00:00

  • Interactions of bioactive lipopeptides, iturin A and surfactin from Bacillus subtilis.

    abstract::The antifungal activity of iturin A and its interaction with erythrocyte membranes were enhanced in the presence of surfactin. The modification of the properties of iturin A was explained by the formation of mixed iturin A-surfactin micelles. Such mixed micelles were easily generated when both lipopeptides were in aqu...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:

    authors: Thimon L,Peypoux F,Maget-Dana R,Roux B,Michel G

    更新日期:1992-10-01 00:00:00

  • Gellan gum biosynthetic enzymes in producing and nonproducing variants of Pseudomonas elodea.

    abstract::A pathway for the synthesis of the repeating tetrasaccharide units in gellan gum from Pseudomonas elodea is proposed. The enzymes presumed to be involved in the synthesis of the activated precursors UDP-glucose, TDP-rhamnose, and UDP-glucuronic acid were detected and assayed in crude cell extracts of the gellan-produc...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:

    authors: Martins LO,Sá-Correia I

    更新日期:1991-12-01 00:00:00

  • Regulatory role of B-cell maturation antigen on the toxic effect of chromium ions on human SaOS-2 osteoblasts.

    abstract::Metal prostheses of artificial joints undergo wear, producing numerous metal particles and ions, such as Cr3+ . Cr3+ is considered a key factor leading to aseptic loosening. Many studies focus on the effect of Cr3+ on osteoblasts; however, little is known about the effect of Cr3+ on the B-cell maturation antigen (BCMA...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.1401

    authors: Li DH,Yang Q,Zhou JS,Zhang ZW,Miao MY,Yang SS,Xu WD

    更新日期:2017-09-01 00:00:00

  • Pattern of growth and respiratory activity of Saccharomyces cerevisiae (baker's yeast) cells growing entrapped in an insolubilized gelatin gel.

    abstract::Yeast cells immobilized by entrapment in an insolubilized gelatin gel have been investigated with regard to their pattern of growth and respiratory activity. Mass-transfer resistance offered by the matrix and growth of the entrapped cells determine a gradient of nutrients throughout the gel which is responsible for bo...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:

    authors: Parascandola P,de Alteriis E

    更新日期:1996-02-01 00:00:00

  • Melamine binding with arachidonic acid binding sites of albumin is a potential mechanism for melamine-induced inflammation.

    abstract::Melamine adulteration of food is a public health concern. It has been seen that melamine causes disease in many organs. Melamine-induced kidney disease is a well-recognized clinicopathological entity. Inflammation is thought to be important in melamine-induced pathology. Melamine is expected to bind with albumin becau...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.1512

    authors: Rajpoot M,Bhattacharyya R,Banerjee D,Sharma A

    更新日期:2017-07-01 00:00:00

  • Vitamin C partially reversed some biochemical changes produced by vitamin E deficiency.

    abstract::Feeding a basal diet free of vitamins E and C to weanling male rats for 8 months resulted in biochemical changes characteristic of vitamin E deficiency. These included increased liver thiobarbituric acid values; decreased blood GSH levels, plasma vitamin E levels, and glutathione peroxidase activities; and increased a...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:

    authors: Chen LH,Thacker RR

    更新日期:1986-02-01 00:00:00

  • Prokaryotic expression, purification and characterization of a novel pro-apoptosis protein hPNAS-4.

    abstract::Recently, hPNAS-4 [human PNAS-4; also known as C1orf121 (Genbank accession number NP-057160)] was reported to be a novel pro-apoptotic protein in mammalian cells. However, at present there is little information about hPNAS-4 and its molecular mechanisms. In our present studies, the hPNAS-4 gene was first cloned into t...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20090111

    authors: Deng H,Jiang Q,Liang S,Yan F,Hou S,Qian Z,Li J,Wen Y,Yang J,Wei Y

    更新日期:2010-02-11 00:00:00

  • Characterization, subsite mapping and partial amino acid sequence of glucoamylase from the filamentous fungus Trichoderma reesei.

    abstract::The pH optimum, temperature-dependence, thermal stability, substrate specificity and subsite affinities of the 66 kDa, pI 4.0 glucoamylase of the filamentous fungus Trichoderma reesei were determined. It had a pH optimum of 5.5 and a temperature optimum (5 min reaction time) of 70 degrees C with soluble starch as subs...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1111/j.1470-8744.1995.tb00333.x

    authors: Fagerström R,Kalkkinen N

    更新日期:1995-04-01 00:00:00

  • Knockdown of GPR137 by RNAi inhibits pancreatic cancer cell growth and induces apoptosis.

    abstract::G-protein-coupled receptors (GPCRs), the largest family of cell-surface molecules involved in a number of biological and pathological processes, have recently emerged as key players in carcinogenesis and cancer progression. Orphan G protein-coupled receptors (oGPCRs) are a group of proteins lacking endogenous ligands....

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.1326

    authors: Cui X,Liu Y,Wang B,Xian G,Liu X,Tian X,Qin C

    更新日期:2015-11-01 00:00:00

  • One-step electrodeposition preparation of polyaniline/f-MWCNTs as electrochemical sensors for detection of 10-hydroxycamptothecine.

    abstract::In our work, one-step electro-deposition method was adopted to produce polyaniline (PANI) and functional multiwalled carbon nanotubes (f-MWCNTs) films on glass carbon electrodes, and the modified electrodes were applied as an electrochemical sensor for determination of 10-hydroxycamptothecine (10-HCPT). The f-MWCNTs w...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.2041

    authors: Meng L,Dai J,Rong S,Gao H,Zhang Z,Zhang Y,Chang D,Pan H

    更新日期:2020-09-24 00:00:00

  • Angiotensin-converting enzyme inhibitory and antioxidant activities of enzymatically synthesized phenolic and vitamin glycosides.

    abstract::Amyloglucosidase from Rhizopus mould and beta-glucosidase from sweet almond were employed for the preparation of phenolic and vitamin glycosides of vanillin, N-vanillylnonanamide, DL-dopa, dopamine, curcumin, alpha-tocopherol (vitamin E), pyridoxine (vitamin B(6)), ergocalciferol (vitamin D(2)), thiamin (vitamin B(1))...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20080041

    authors: Charles RE,Ponrasu T,Sivakumar R,Divakar S

    更新日期:2009-03-01 00:00:00

  • A strategic study using mutant-strain entrapment in calcium alginate for the production of Saccharomyces cerevisiae cells with high invertase activity.

    abstract::Entrapped cells and entrapped cells grown inside of a calcium alginate matrix as well as free cells have been investigated using Saccharomyces cerevisiae mutant strains with regard to their pattern of growth and invertase activity. The repression of invertase by glucose and glucose-consumption ability were considered ...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20020101

    authors: Rossi-Alva JC,Rocha-Leão MH

    更新日期:2003-08-01 00:00:00

  • Purification and biochemical characterization of a thermostable, alkaliphilic, extracellular alpha-amylase from Bacillus subtilis DM-03, a strain isolated from the traditional fermented food of India.

    abstract::Bacillus subtilis strain DM-03, which is isolated from starter culture used for the production of alcohol by local Assam tribes, grows optimally at 52-55 degrees C and secretes a significant amount of alpha-amylase at pH 8.0 into the culture media. This alpha-amylase, purified by ion-exchange, gel-filtration and rever...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20040034

    authors: Das K,Doley R,Mukherjee AK

    更新日期:2004-12-01 00:00:00

  • CYP261 enzymes from deep sea bacteria: a clue to conformational heterogeneity in cytochromes P450.

    abstract::We have explored the adaptation of the cytochromes P450 (P450) of deep-sea bacteria to high hydrostatic pressures. Strict conservation of the protein fold and functional importance of protein-bound water make P450 a unique subject for the studies of high-pressure adaptation. Earlier, we expressed and purified a fatty-...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.1083

    authors: Davydov DR,Sineva EV,Davydova NY,Bartlett DH,Halpert JR

    更新日期:2013-01-01 00:00:00

  • Immobilization of Acidithiobacillus ferrooxidans on cotton gauze for biological oxidation of ferrous ions in a batch bioreactor.

    abstract::The ability of Acidithiobacillus ferrooxidans to oxidize ferrous iron has been extensively studied in bioleaching to recover metal resources. Although immobilization of A. ferrooxidans is of great importance to achieve high bioleaching performance in practical application, the reported approaches of immobilization of ...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.1464

    authors: Zhu N,Shi C,Shang R,Yang C,Xu Z,Wu P

    更新日期:2017-09-01 00:00:00

  • Biobutanol production from sugarcane bagasse by Clostridium beijerinckii strains.

    abstract::Acetone-butanol-ethanol (ABE) fermentation was performed with sugarcane bagasse (SCB) hydrolysate using Clostridium beijerinckii strains. A cost-effective SCB medium was developed with no enzymatic hydrolysis and no supplementation of extra carbon source or expensive nitrogen source. One of the C. beijerinckii strains...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1002/bab.1865

    authors: Narayanasamy S,Chan KL,Cai H,Abdul Razak AHB,Tay BK,Miao H

    更新日期:2020-09-01 00:00:00

  • Preparation of active hybrid enzymes composed of the native and chemically inactivated aspartase subunits from Escherichia coli.

    abstract::The hybridization of the native and chemically inactivated aspartase from Escherichia coli was studied. Preparations of the tetrameric enzyme obtained by mixing the native and N-ethylmaleimide (NEM)-inactivated aspartase in 4 M guanidine-HCl followed by 51-fold dilution at room temperature retained catalytic activity....

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:

    authors: Imaishi H,Yumoto N,Tokushige M

    更新日期:1990-04-01 00:00:00

  • Argatroban-coupled Affi-Gel matrix for the purification of thrombin from plasma.

    abstract::Sometimes it is necessary to obtain thrombin from limited amounts of human plasma for laboratory assay. None of the available purification methods easily deals with this subject. The procedure described in the present paper uses a readily available pharmaceutical agent, argatroban, to construct an affinity matrix. Arg...

    journal_title:Biotechnology and applied biochemistry

    pub_type: 杂志文章

    doi:10.1042/BA20050057

    authors: Lefkowitz JB

    更新日期:2005-10-01 00:00:00