Abstract:
:During the screening for tyrosine phenol-lyase-producing thermophiles, we isolated an obligatory symbiotic thermophile, Symbiobacterium sp. SC-1, which grew only in coculture with Bacillus sp. SK-1. A gene encoding thermostable tyrosine phenol-lyase (TPL) was cloned from the genomic DNA of the Symbiobacterium sp. SC-1 and the nucleotide sequence of the TPL structural gene was determined. The gene consists of 1374 base pairs encoding a polypeptide of 458 amino acid residues; the molecular mass of the enzyme subunit is estimated to be 52,196 Da. The structural gene of TPL was amplified by PCR, blunt-ended, and ligated into the NcoI-HindIII site of plasmid pTrc99A to construct an expression vector for the overproduction of the thermostable TPL. The level of thermostable TPL production was about 15% of the total soluble proteins of Escherichia coli extract. The enzyme was purified to homogeneity from the E. coli extract with an overall yield of 48%.
journal_name
Protein Expr Purifjournal_title
Protein expression and purificationauthors
Lee SG,Hong SP,Choi YH,Chung YJ,Sung MHdoi
10.1006/prep.1997.0792subject
Has Abstractpub_date
1997-12-01 00:00:00pages
263-70issue
3eissn
1046-5928issn
1096-0279pii
S1046-5928(97)90792-0journal_volume
11pub_type
杂志文章abstract::Transcription factor HIF-1 is a key regulator in cellular adaptation to hypoxia. HIF prolyl hydroxylases (PHDs) control HIF-1 accumulation by hydroxylation dependent on molecular oxygen. Due to this regulation, PHDs have been pointed out as potential drug targets. We have purified catalytically active human PHD3 after...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.02.018
更新日期:2007-07-01 00:00:00
abstract::The second enzyme of the naphthalene degradation pathway in Pseudomonas putida G7 is NahB, a dehydrogenase that converts cis-1,2-dihydroxy-1,2-dihydronaphthalene to 1,2-dihydroxynaphthalene. We report the cloning, optimization of expression, purification, kinetic studies and preliminary structural characterization of ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2017.01.005
更新日期:2017-04-01 00:00:00
abstract::Expressed protein libraries are becoming a critical tool for new target discovery in the pharmaceutical industry. In order to get the most meaningful and comprehensive results from protein library screens, it is essential to have library proteins in their native conformation with proper post-translation modifications....
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.05.011
更新日期:2011-09-01 00:00:00
abstract::Expression and purification of human beta-secretase (BACE1) in bacteria have been plagued with issues concerning solubility, inhomogeneous N-terminus, and lack of enzymic activity. Several forms of the mature human BACE1 have been expressed in Escherichia coli with different N-terminal extensions and without the C-ter...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2003.12.014
更新日期:2004-04-01 00:00:00
abstract::Human dermcidin, an anionic antimicrobial peptide expressed in the pons of the brain and the sweat glands, displays antimicrobial activity against pathogenic microorganisms such as Staphylococcus aureus and Candida albicans. Here, we describe the recombinant production of a 48 amino acid dermcidin variant with C-termi...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.07.002
更新日期:2006-02-01 00:00:00
abstract::The present report documents an improved enzyme assay method for the mammalian L-alanine:4,5-dioxovalerate transaminase which is of significant utility in work with crude tissue homogenates, cell cultures, or purified enzyme preparations. We also describe a new and rapid purification procedure for this enzyme from rat...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1994.1072
更新日期:1994-12-01 00:00:00
abstract::Streptococcal protein G has been used extensively for the purification of antibodies using the interaction of the Fc region with protein G. Many antibodies also interact with protein G through a low-affinity binding site for the Fab region. The exploitation of this low-affinity interaction for the purification of Fab'...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(05)80037-3
更新日期:1992-10-01 00:00:00
abstract::Production of VCSM13 phage displaying a high density of CD147 ectodomain (CD147Ex) was achieved when culturing conditions were modulated. A phagemid expressing CD147Ex was constructed and used to produce phage display CD147Ex gpVIII fusion protein in TG1 Escherichia coli. Displaying of CD147Ex via gpVIII was successfu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2003.08.019
更新日期:2003-12-01 00:00:00
abstract::Our earlier studies have demonstrated that the 35 kDa isoform of Translocated promoter region protein (Tpr) of Rattus norvegicus was able to augment c-jun transcription efficiently. Identification of direct targets that may in part downregulate c-jun transcription might prove to be an ideal target to curtail the proli...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.01.001
更新日期:2011-05-01 00:00:00
abstract::In order to produce sufficient quantities of fibroblast growth factor-saporin (rFGF-2-SAP) mitotoxin for preclinical evaluation in models of diseases such as cancer and restenosis, we have undertaken the large-scale expression, purification, and characterization of the recombinant molecule. The fusion gene encoding rF...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0079
更新日期:1996-08-01 00:00:00
abstract::Proteomics has become a major focus as researchers attempt to understand the vast amount of genomic information. Protein complexity makes identifying and understanding gene function inherently difficult. The challenge of studying proteins in a global way is driving the development of new technologies for systematic an...
journal_title:Protein expression and purification
pub_type: 杂志文章,评审
doi:10.1006/prep.2001.1465
更新日期:2001-07-01 00:00:00
abstract::An Escherichia coli expression system that exploits the bacterial alkaline phosphatase (PhoA) signal sequence to translocate recombinant human epidermal growth factor (hEGF) to the periplasm was used to evaluate how changes in the composition and sequence of amino acids near the PhoA-hEGF junction influence the peripl...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1997.0741
更新日期:1997-08-01 00:00:00
abstract::We describe an efficient strategy to produce high-quality proteins by using a single large IMAC chromatography column and enzymatic His-tag removal via the TAGZyme system in pilot scale. Numerous quality assays demonstrated a high purity of the final product, the human cytokine Interleukin-1beta (IL-1beta). The protei...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2007.09.019
更新日期:2008-02-01 00:00:00
abstract::CD23, a 45-kDa type II membrane glycoprotein present on B cells, monocytes, and other human immune cells, is a low-affinity receptor for IgE. The extracellular region of the membrane-bound human CD23 is processed into at least four soluble (s) CD23 forms, with apparent molecular masses of 37, 33, 29, and 25 kDa. High ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1448
更新日期:2001-07-01 00:00:00
abstract::Quinolinate synthetase catalyzes the second step of the de novo biosynthetic pathway of pyridine nucleotide formation. In particular, quinolinate synthetase is involved in the condensation of dihydroxyacetone phosphate and iminoaspartate to form quinolinic acid. To study the mechanism of action, the specificity of the...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1153
更新日期:2000-02-01 00:00:00
abstract::Integrated studies that address proteins structure and function in the new era of systems biology and genomics often require the application of high-throughput approaches for parallel production of many different purified proteins from the same organism. Cytochromes c-electron transfer proteins carrying one or more he...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.06.014
更新日期:2008-11-01 00:00:00
abstract::Disulfide bonds are normally formed after a polypeptide has been exported from the reducing environment of the cytoplasm into a more oxidizing compartment, such as the bacterial periplasm. Recently, we showed that in Escherichia coli trxB gor mutants, in which the reduction of thioredoxin and glutathione is impaired, ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1520
更新日期:2001-11-01 00:00:00
abstract::The guanine nucleotide exchange factor EF-1beta gene from the thermoacidophilic archaeon Sulfolobus solfataricus (SsEF-1beta) was amplified by PCR and cloned into the pT7-7 expression vector. One of four selected clones harbored the T160C nucleotide substitution leading to the Y54H amino acid change in a hydrophobic r...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1997.0806
更新日期:1998-02-01 00:00:00
abstract::A stable mammalian cell line expressing highly active bovine interferon-gamma (BoIFN-γ) was generated using Flp recombinase-mediated integration. This recombinant 293 cell line (B1) efficiently secreted FLAG-tagged BoIFN-γ protein into the culture supernatant, as determined by ELISA and Western blot. The recombinant B...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2014.04.012
更新日期:2014-07-01 00:00:00
abstract::Previously, the lysozyme gene of the Klebsiella phage K11 was partially sequenced in our lab. Using the sequence information the lysozyme gene of the Klebsiella phage K11 was amplified and cloned using the polymerase chain reaction of the pfu DNA polymerase. The nucleotide sequence of phage K11 lysozyme gene was deter...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.03.026
更新日期:2005-07-01 00:00:00
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
更新日期:2018-12-01 00:00:00
abstract::We previously reported the expression and purification of recombinant p68 RNA helicase in a bacterial expression system. The recombinant p68 is an RNA-dependent ATPase and ATP-dependent RNA helicase. In the process of characterizing the ATPase and RNA unwinding activities of the recombinant p68, we observed that the b...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.02.004
更新日期:2004-06-01 00:00:00
abstract::A tremendous increase has taken place over the last decades in the biochemical and clinical use of antibodies. Unfortunately, the constantly growing demand has not been matched by a corresponding easy access to pure immunoglobulin, as purification procedures tend to be either laborious, expensive, or inefficient. We p...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00130-x
更新日期:2003-08-01 00:00:00
abstract::Human pancreatic ribonuclease (HP-RNase) has considerable promise as a therapeutic agent. Structure-function analyses of HP-RNase have been impeded by the difficulty of obtaining the enzyme from its host. Here, a gene encoding HP-RNase was designed, synthesized, and inserted into two expression vectors that then direc...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0036
更新日期:1996-05-01 00:00:00
abstract::The human iron-binding protein lactoferrin (hLf) has been implicated in a number of important physiological pathways, including those regulating immune function and tumor growth. In an effort to develop an efficient system for production of recombinant hLf (rhLf) that is structurally and functionally equivalent to the...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.01.003
更新日期:2009-05-01 00:00:00
abstract::The full-length human acetyl-CoA carboxylase 1 (ACC1) was expressed and purified to homogeneity by two separate groups (Y.G. Gu, M. Weitzberg, R.F. Clark, X. Xu, Q. Li, T. Zhang, T.M. Hansen, G. Liu, Z. Xin, X. Wang, T. McNally, H. Camp, B.A. Beutel, H.I. Sham, Synthesis and structure-activity relationships of N-{3-[2...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.11.021
更新日期:2007-05-01 00:00:00
abstract::A cysteine-rich (approximately 20%), low molecular weight (MW 6 kDa) polypeptide has been isolated from the Korean blood-sucking leech, Hirudo nipponia. From its amino acid composition and N-terminal amino acid sequence analysis, the new protein is similar to granulin (or epithelin), and so it has been named leech gra...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1999.1077
更新日期:1999-07-01 00:00:00
abstract::Mitochondria-mediated apoptosis (programmed cell death) involves a sophisticated signaling and regulatory network that is regulated by the Bcl-2 protein family. Members of this family have either pro- or anti-apoptotic functions. An important pro-apoptotic member of this family is the cytosolic Bax. This protein is cr...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2019.02.004
更新日期:2019-06-01 00:00:00
abstract::Homocysteine alpha,gamma-lyase from the anaerobic protozoan parasite Trichomonas vaginalis has been cloned from genomic DNA using PCR methods and expressed in Escherichia coli with a vector containing the T7 promoter. The recombinant homocysteine alpha,gamma-lyase (rHCYase) is expressed as the major protein in the hos...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1998.0955
更新日期:1998-11-01 00:00:00
abstract::Deoxynivalenol (DON), a mycotoxin produced by several Fusarium species, is a worldwide contaminant of food and feedstuffs. The DON-specific single-chain variable fragment (scFv) antibody was produced in recombinant Escherichia coli. The variable regions of the heavy chain (V(H)) and light chain (V(L)) cloned from the ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2003.12.008
更新日期:2004-05-01 00:00:00