当前位置： SCI文献检索 > PROTEIN EXPRESSION AND PURIFICATION期刊下所有文献 > Overexpression of salt-tolerant glutaminase from Micrococcus luteus K-3 in Escherichia coli and its purification.

Overexpression of salt-tolerant glutaminase from Micrococcus luteus K-3 in Escherichia coli and its purification.

Abstract：

:A high-expression plasmid, pKSGHE3-1, containing the salt-tolerant glutaminase (EC 3.5.1.2) from marine bacterium Micrococcus luteus K-3 was constructed. pKSGHE3-1 was made by inserting the DNA fragment (1.43 kb) containing the structural gene synthesized by polymerase chain reaction into the downstream region of the tac promoter of expression vector pKK223-3. The translational start codon was located 10 bases downstream of the Shine-Dalgarno sequence (AGGA) of pKK223-3. Escherichia coli JM109 transformed with pKSGHE3-1 exhibited more than 190-fold higher glutaminase activity than M. luteus K-3 under optimal culture conditions. The enzyme was purified to homogeneity through three column chromatography steps with a final yield of 17.1%. The recombinant enzyme showed the same enzymatic properties, including salt tolerance, as those of M. luteus K-3. This glutaminase expression system allows the production of sufficient quantities of glutaminase for basic structure-function studies including chemical modification and future X-ray crystallization analysis.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Nandakumar R,Wakayama M,Nagano Y,Kawamura T,Sakai K,Moriguchi M

doi

10.1006/prep.1998.1005

keywords:

subject

Has Abstract

pub_date

1999-03-01 00:00:00

pages

155-61

issue

2

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(98)91005-1

journal_volume

15

pub_type

杂志文章

相关文献

PROTEIN EXPRESSION AND PURIFICATION文献大全
  • Solution studies of recombinant human stromal-cell-derived factor-1.

    abstract：:Stromal-cell-derived factor-1 (SDF-1alpha) is an 8-kDa chemokine that is constitutively expressed in bone-marrow-derived stromal cells and has been identified as a ligand for the CXCR4 receptor. We produced the chemokine recombinantly as methionine-SDF-1alpha in Escherichia coli without the leader peptide sequence. Th...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2001.1402

    authors： Holmes WD,Consler TG,Dallas WS,Rocque WJ,Willard DH

    更新日期：2001-04-01 00:00:00

  • Expression, purification and preliminary characterization of glucagon receptor extracellular domain.

    abstract：:Glucagon is a pancreatic hormone that plays pivotal roles in regulating glucose homeostasis and metabolism. Glucagon exerts its action by binding to its receptor, glucagon receptor (GCGR), one of class B G-protein coupled receptors (GPCRs). Diabetes is a bihormonal disease in which excessive glucagon secretion is a ma...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2013.04.004

    authors： Wu L,Zhai Y,Lu J,Wang Q,Sun F

    更新日期：2013-06-01 00:00:00

  • Immediate-early baculovirus vectors for foreign gene expression in transformed or infected insect cells.

    abstract：:Baculovirus expression vectors are used routinely for foreign gene expression and are under intense development as improved biological pesticides. Conventional baculovirus expression vectors are recombinant viruses that can express a foreign gene in insect cells under the control of the polyhedrin promoter, which prov...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1996.0092

    authors： Jarvis DL,Weinkauf C,Guarino LA

    更新日期：1996-09-01 00:00:00

  • Transmembrane-sequence-dependent overexpression and secretion of glycoproteins in Saccharomyces cerevisiae.

    abstract：:Protein expression using the secretory pathway in Saccharomyces cerevisiae can lead to high amounts of overexpressed and secreted proteins in culture supernatants in a short period of time. These post-translational modified expression products can be purified up to >90% in a single step. The overexpression and secreti...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.2000.1337

    authors： Schuster M,Wasserbauer E,Aversa G,Jungbauer A

    更新日期：2001-02-01 00:00:00

  • Purification and biochemical characterization of a recombinant mouse seminal vesicle trypsin inhibitor produced in Escherichia coli.

    abstract：:Escherichia coli cells were transformed with an expression vector constructed by inserting a DNA fragment encoding a Kazal-type trypsin inhibitor from mouse seminal vesicle into pGEX-2. The cloned cells were able to produce a high yield of a chimeric polypeptide made by fusing the trypsin inhibitor to glutathione S-tr...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1994.1003

    authors： Lai ML,Li SH,Chen YH

    更新日期：1994-02-01 00:00:00

  • Expression of Pisum sativum SAD polypeptides in production hosts and in planta: tetrameric organization of the protein.

    abstract：:In Pisum sativum, the short-chain alcohol dehydrogenase-like protein (SAD) gene family consists of at least three members (SAD-A, -B, and -C). Expression of two of these genes (SAD-A and -C) in Escherichia coli or Pichia pastoris resulted in full-length soluble proteins. Purified SAD-A was used as antigen for antibody...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2008.09.004

    authors： Scherbak N,Brosché M,Ala-Häivälä A,Strid H,Ohrfelt A,Nilsson F,Strid A

    更新日期：2009-01-01 00:00:00

  • Purification of recombinant nacre-associated mineralization protein AP7 fused with maltose-binding protein.

    abstract：:Formation of biominerals often involves specific proteins that modulate the process of matrix assembly, nucleation, and crystal growth. AP7 is an aragonite-associated protein of 7 kDa and is intrinsically disordered. The structural disorder of AP7 makes it very difficult to express in Escherchiacoli. In this work, we ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2014.05.002

    authors： Huang YC,Chang HH,Mou Y,Chi P,Chan JC,Luo SC

    更新日期：2014-08-01 00:00:00

  • High-level expression and purification of active scorpion long-chain neurotoxin BjαIT from Pichia pastoris.

    abstract：:As an insect-selective neurotoxin, scorpion long-chain BjαIT is a promising prospect for insecticidal application; however, the difficulty of obtaining natural BjαIT represents the major obstacle preventing analysis of its insecticidal activity against agricultural insect pests. Here, we screened recombinant Pichia pa...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2018.07.013

    authors： Li H,Xia Y

    更新日期：2018-12-01 00:00:00

  • Heme content of recombinant catalase from Psychrobacter sp. T-3 altered by host Escherichia coli cell growth conditions.

    abstract：:The catalase gene of Psychrobacter sp. T-3 was cloned, and the gene product (PktA) was overexpressed in Escherichia coli. The specific activity of the purified PktA was slightly lower than that of the native purified enzyme obtained from Psychrobacter sp. T-3. Spectrophotometric measurements of the purified enzymes su...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2008.03.016

    authors： Kimoto H,Matsuyama H,Yumoto I,Yoshimune K

    更新日期：2008-06-01 00:00:00

  • Expression, purification, and biochemical characterization of enteroaggregative Escherichia coli heat-stable enterotoxin 1.

    abstract：:Enteroaggregative Escherichia coli (EAEC) heat-stable enterotoxin 1 (EAST1) is a 4.1k Da protein originally discovered in EAEC but known to be scattered in other diarrheagenic E. coli as well, possibly causing diarrhea in humans and animals. We report for the first time a method to express and purify EAST1 using the G...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2003.09.008

    authors： Ménard LP,Lussier JG,Lépine F,Paiva de Sousa C,Dubreuil JD

    更新日期：2004-02-01 00:00:00

  • Process development for production and purification of the Schistosoma mansoni Sm14 antigen.

    abstract：:The trematode Schistosoma mansoni Sm14 antigen was expressed in the yeast Pichia pastoris and secreted into the culture medium at yields of approximately 250 mg L-1. Sm14 belongs to a family of fatty-acid binding proteins and appears to play an important role in uptake, transport, and compartmentalization of lipids in...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2017.04.002

    authors： Damasceno L,Ritter G,Batt CA

    更新日期：2017-06-01 00:00:00

  • The screening of expression and purification conditions for replicative DNA polymerase associated B-subunits, assignment of the exonuclease activity to the C-terminus of archaeal pol D DP1 subunit.

    abstract：:The B-subunits of replicative DNA polymerases belong to the superfamily of calcineurin-like phosphoesterases and are conserved from Archaea to humans. Recently we and others have shown that the B-subunit (DP1) of the archaeal family D DNA polymerase is responsible for proofreading 3'-5' exonuclease activity. The simil...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.05.002

    authors： Jokela M,Raki M,Heikkinen K,Sepponen K,Eskelinen A,Syväoja JE

    更新日期：2005-09-01 00:00:00

  • Cloning, purification, and refolding of human paraoxonase-3 expressed in Escherichia coli and its characterization.

    abstract：:Human paraoxonase (hPON3) is a high density lipoprotein-related glycoprotein with multi-enzymatic properties and antioxidant activity which is proposed to participate in the prevention of low density lipoprotein (LDL) oxidation. In this study, hPON3 gene was amplified from Human Fetal Liver Marathon-Ready cDNA and exp...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.07.021

    authors： Lu H,Zhu J,Zang Y,Ze Y,Qin J

    更新日期：2006-03-01 00:00:00

  • Expression, purification and characterization of cold shock protein A of Corynebacterium pseudotuberculosis.

    abstract：:The gram-positive bacterium Corynebacterium pseudotuberculosis is the causative agent of different diseases that cause dramatically reduced yields of wool and milk, and results in weight loss, carcass condemnation and also death mainly in sheep, equids, cattle and goats and therefore globally results in considerable e...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.04.006

    authors： Lindae A,Eberle RJ,Caruso IP,Coronado MA,de Moraes FR,Azevedo V,Arni RK

    更新日期：2015-08-01 00:00:00

  • Rapid one-step isolation of mouse liver catalase by immobilized metal ion affinity chromatography.

    abstract：:A novel and rapid procedure for the isolation of catalase from mouse liver, after prior treatment with the peroxisome proliferator perfluorooctanoic acid was developed using immobilized metal ion affinity chromatography involving chelation with zinc ions. The purification developed is simple, rapid (requiring 3 hours ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1997.0827

    authors： Yang Q,DePierre JW

    更新日期：1998-03-01 00:00:00

  • Purification and characterization of fibrinolytic enzyme from cultured mycelia of Armillaria mellea.

    abstract：:A fibrinolytic enzyme was purified from the cultured mycelia of Armillaria mellea by ion-exchange chromatography followed by gel filtration, and was designated A. mellea metalloprotease (AMMP). The purification protocol resulted in a 627-fold purification of the enzyme, with a final yield of 6.05%. The apparent molecu...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2005.05.004

    authors： Lee SY,Kim JS,Kim JE,Sapkota K,Shen MH,Kim S,Chun HS,Yoo JC,Choi HS,Kim MK,Kim SJ

    更新日期：2005-09-01 00:00:00

  • Overexpression, oxidative refolding, and zinc binding of recombinant forms of the murine S100 protein MRP14 (S100A9).

    abstract：:Recombinant murine MRP14 (mMRP14) was produced in Escherichia coli using the pGEX expression system. The mass of fusion protein, by electrospray ionization-mass spectrometry (ESI/MS), was 39,213 Da which compares well with the theoretical mass (39,210.4 Da). Thrombin digestion of fusion protein was expected at a clone...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1998.1015

    authors： Raftery MJ,Collinson L,Geczy CL

    更新日期：1999-03-01 00:00:00

  • First characterization of extremely halophilic 2-deoxy-D-ribose-5-phosphate aldolase.

    abstract：:2-Deoxy-d-ribose-5-phosphate aldolase (DERA) catalyzes the aldol reaction between two aldehydes and is thought to be a potential biocatalyst for the production of a variety of stereo-specific materials. A gene encoding DERA from the extreme halophilic archaeon, Haloarcula japonica, was overexpressed in Escherichia col...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2016.05.009

    authors： Ohshida T,Hayashi J,Satomura T,Kawakami R,Ohshima T,Sakuraba H

    更新日期：2016-10-01 00:00:00

  • Expression and purification of Arg196 and Lys272 mutants of mevalonate kinase from Methanococcus jannaschii.

    abstract：:In microorganisms and plants, mevalonate kinase is involved in the biosynthesis of isoprenoid derivatives, one of the largest groups of natural products. We subcloned the gene of mevalonate kinase from Methanococcus jannaschii into a bacterial expression vector pLM1 with six continuous histidine codons attached to the...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/s1046-5928(03)00101-3

    authors： Chu X,Liu X,Yau M,Leung YC,Li D

    更新日期：2003-08-01 00:00:00

  • Cloning, expression, purification and functional characterization of the oligomerization domain of Bcr-Abl oncoprotein fused to the cytoplasmic transduction peptide.

    abstract：:Protein-based cellular therapeutics have been limited by getting molecules into cells and the fact that many proteins require accurate cellular localization for function. Cytoplasmic transduction peptide (CTP) is a newly designed transduction peptide that carries molecules across the cell membrane with a preference to...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2008.10.023

    authors： Huang SF,Liu DB,Zeng JM,Xiao Q,Luo M,Zhang WP,Tao K,Wen JP,Huang ZG,Feng WL

    更新日期：2009-04-01 00:00:00

  • Expression, purification, and characterization of human malonyl-CoA decarboxylase.

    abstract：:The recombinant human malonyl-CoA decarboxylase (hMCD) was overexpressed in Escherichia coli with and without the first 39 N-terminal amino acids via a cleavable MBP-fusion construct. Proteolytic digestion using genenase I to remove the MBP-fusion tag was optimized for both the full length and truncated hMCD. The apo-...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2003.11.023

    authors： Zhou D,Yuen P,Chu D,Thon V,McConnell S,Brown S,Tsang A,Pena M,Russell A,Cheng JF,Nadzan AM,Barbosa MS,Dyck JR,Lopaschuk GD,Yang G

    更新日期：2004-04-01 00:00:00

  • Expression and purification recombinant antihypertensive peptide ameliorates hypertension in rats with spontaneous hypertension.

    abstract：:A highly efficient Escherichia coli expression system was established to obtain an appreciable quantity of antihypertensive peptide. The DNA-coding sequence for the Gly-Val-Tyr-Pro-His-Lys peptide was chemically synthesized and linked to form a ten-copy in tandem. It was cloned into the vector pET-15b and expressed in...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2015.05.001

    authors： Wang XL,Ma SN,Yuan YH,Ding Y,Li DS

    更新日期：2015-09-01 00:00:00

  • Low binding affinity and reduced complement-dependent cell death efficacy of ofatumumab produced using a plant system (Nicotiana benthamiana L.).

    abstract：:The plant protein production system is a platform that can not only reduce production costs but also produce monoclonal antibodies that do not have the risk of residual proteins from the host. However, due to the difference between post-translational processes in plants and animals, there may be a modification in the ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2019.03.004

    authors： Jin N,Lee JW,Heo W,Ryu MY,So MK,Ko BJ,Kim HY,Yoon SM,Lee J,Kim JY,Kim WT

    更新日期：2019-07-01 00:00:00

  • Identification of UDP-linked murein precursors as contaminants in recombinant proteins of low molecular weight.

    abstract：:The A(280)/A(260) ratio of a purified protein is frequently used as an indication of the purity of the preparation with respect to nucleic acids. We show here that for low-molecular-weight recombinant proteins purified from Escherichia coli, a low A(280)/A(260) ratio can also result from contamination with UDP-linked ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1999.1145

    authors： Ram MK,Andrade LJ,Phillips TB,van Schravendijk MR

    更新日期：1999-11-01 00:00:00

  • Fed-batch production of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in soluble form in Escherichia coli and its purification and characterization.

    abstract：:Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anticancer agent. The aim of this study is to produce large quantities of highly pure and bioactive recombinant human TRAIL. Here, TRAIL was expressed in soluble form by pH-stat fed-batch cultivation and purified using a rapid and simple tw...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2016.06.007

    authors： Li P,Gu Q,Wu X

    更新日期：2016-10-01 00:00:00

  • Efficient bacterial expression of recombinant potato mop-top virus non-structural triple gene block protein 1 modified by progressive deletion of its N-terminus.

    abstract：:To obtain strong bacterial expression of proteins that seem to be hard to express in bacteria or are highly toxic for bacteria, it is possible to create a palette of similar constructs, differing only by several nucleotides, gradually deleted from the full-length clone by exonuclease III. When a construct is equipped ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2004.12.013

    authors： Pecenková T,Filigarová M,Cerovská N

    更新日期：2005-05-01 00:00:00

  • Expression, purification and characterization of TMCO1 for structural studies.

    abstract：:Transmembrane and coiled-coil domains 1 (TMCO1) has a highly conserved amino acid sequence among species, indicating a critical role of TMCO1 in cell physiology. The deficiency of TMCO1 in humans is associated with cerebrofaciothoracic dysplasia (CFTD), glaucoma, osteogenesis and the occurrence of cancer. TMCO1 was re...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2020.105803

    authors： Zhang N,Tang M,Wen M,Cao Y,OuYang B

    更新日期：2021-03-01 00:00:00

  • Isolation, overexpression, and biochemical characterization of the two isoforms of glutamic acid decarboxylase from Escherichia coli.

    abstract：:Escherichia coli glutamic acid decarboxylase is a pyridoxal phosphate-dependent enzyme that catalyzes the alpha-decarboxylation of glutamate to yield 4-amino-butyrate and CO2. The E. coli chromosome contains two genes encoding for this enzyme, gadA and gadB, which map at distinct loci. Their protein products differ in...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1996.0121

    authors： De Biase D,Tramonti A,John RA,Bossa F

    更新日期：1996-12-01 00:00:00

  • Human cysteine-rich intestinal protein: cDNA cloning and expression of recombinant protein and identification in human peripheral blood mononuclear cells.

    abstract：:Cysteine-rich intestinal protein (CRIP) is a small, 8.5-kDa protein with one double zinc-finger motif called a LIM domain. It is very abundant in intestine and some immune cells in rodents, and expression is influenced by development and the immune response. We have cloned a human CRIP cDNA from human small intestine ...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1006/prep.1996.0709

    authors： Khoo C,Blanchard RK,Sullivan VK,Cousins RJ

    更新日期：1997-04-01 00:00:00

  • Improved inducible expression of Bacillus naganoensis pullulanase from recombinant Bacillus subtilis by enhancer regulation.

    abstract：:Pullulanase is crucial to the specific hydrolysis of branch points in amylopectin and is generally employed as an important enzyme in the starch-processing industry. Recombinant Bacillus subtilis that employs an inducible promoter would be a suitable candidate for pullulanase expression because of its safety and contr...

    journal_title：Protein expression and purification

    pub_type： 杂志文章

    doi：10.1016/j.pep.2018.03.012

    authors： Deng Y,Nie Y,Zhang Y,Wang Y,Xu Y

    更新日期：2018-08-01 00:00:00