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::Arginase (EC 3.5.3.1; L-arginine amidinohydrolase) is a key enzyme of the urea cycle that catalyses the conversion of arginine to ornithine and urea, which is the final cytosolic reaction of urea formation in the mammalian liver. The recombinant strain of the yeast Saccharomyces cerevisiae that is capable of overprodu...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.09.001
更新日期:2012-01-01 00:00:00
abstract::Protein asparagine (N)-linked glycosylation is a post-translational modification that occurs in the endoplasmic reticulum; it plays an important role in protein folding, oligomerization, quality control, sorting, and transport. Accordingly, disorders of glycosylation may affect practically every organ system. Dehydrod...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2017.02.001
更新日期:2017-04-01 00:00:00
abstract::Methionine aminopeptidases (MetAPs), ubiquitous enzymes that play an important role in nascent protein maturation, have been recognized as attractive targets for the development of drugs against pathogenic protozoa including Plasmodium spp. Here, we characterized partial biochemical properties of a type I MetAP of Pla...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.01.003
更新日期:2015-04-01 00:00:00
abstract::Recombinant barley high pI alpha-glucosidase was produced by high cell-density fermentation of Pichia pastoris expressing the cloned full-length gene. The gene was amplified from a genomic clone and exons (coding regions) were assembled by overlap PCR. The resulting cDNA was expressed under control of the alcohol oxid...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2005.10.008
更新日期:2006-03-01 00:00:00
abstract::Tyrosine hydroxylase is the rate-limiting step in the synthesis of dopamine and is tightly regulated. Previous studies have shown it to be covalently modified and potently inhibited by 3,4-dihydroxyphenylacetaldehyde (DOPAL), an endogenous neurotoxin via dopamine catabolism which is relevant to Parkinson's disease. In...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2012.05.007
更新日期:2012-08-01 00:00:00
abstract::The human melanocortin 4 receptor (MC4r) was successfully expressed in Sf9 cells using the baculovirus infection system. N- and C-terminally His-tagged receptors generated B(max) values of 14 and 23 pmol receptor/mg membrane protein, respectively. The highest expression level obtained with the C-terminally His-tagged ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2004.06.018
更新日期:2004-10-01 00:00:00
abstract::Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination, and repair in bacteria, archaea, and eukarya. We report here the identification, expression, and purification of the SSB-like proteins of the thermophilic bacteria Thermus thermophilus and T. aquaticus. The nucleotide (...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00504-1
更新日期:2002-10-01 00:00:00
abstract::Dps, found in many eubacterial and archaebacterial species, appears to protect cells from oxidative stress and/or nutrient-limited environment. Dps has been shown to accumulate during the stationary phase, to bind to DNA non-specifically, and to form a crystalline structure that compacts and protects the chromosome. O...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2008.01.015
更新日期:2008-06-01 00:00:00
abstract::The human PRODH gene has been shown to have unique roles in regulating cell survival and apoptotic pathways and it has been related to velocardiofacial syndrome/DiGeorge syndrome and increased susceptibility to schizophrenia. It encodes for the flavoprotein proline oxidase (PO), which catalyzes the conversion of l-pro...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2012.01.021
更新日期:2012-04-01 00:00:00
abstract::A process for bacterial expression and purification of the recombinant major wasp allergen Antigen 5 (Ves v 5) was developed to produce protein for diagnostic and therapeutic applications for type 1 allergic diseases. Special attention was focused on medium selection, fermentation conditions, and efficient refolding p...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2006.01.009
更新日期:2006-06-01 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::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::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
abstract::Pen b 26 is one of the allergens produced by Penicillium brevicompactum which is one of the most prevalent in door airborne fungi and a major source of respiratory problems, including asthma. Pen b 26 wa scloned and expressed as an N-terminal as well as a C-terminal His6 tagged fusion protein in Escherichia coli. This...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2009.01.004
更新日期:2009-05-01 00:00:00
abstract::The cDNA encoding the 65-kDa subunit of malic enzyme from Ascaris suum was cloned into the bacterial expression vector pKK223-3 and overproduced in Escherichia coli. A protein with a subunit molecular mass of 65,000 was expressed at a level of up to 3% of the total soluble protein in JM109, as judged by SDS-PAGE. The ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0705
更新日期:1997-06-01 00:00:00
abstract::The entire extracellular domain of the human heat-stable enterotoxin (ST) receptor as well as a truncated N-terminal domain were cloned as glutathione S-transferase fusion proteins and expressed in Escherichia coli. The recombinant fusion proteins were purified from both the cytosol and the inclusion body fractions by...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1996.0087
更新日期:1996-09-01 00:00:00
abstract::The erm proteins confer resistance to the MLS (macrolide-lincosamide-streptogramin B) antibiotics in various microorganisms, including pathogens, through dimethylation of a single adenine residue (A2085: Bacillus subtilis coordinate) of the 23S rRNA to reduce the affinity of antibiotics, thereby enabling the cells to ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2002.1621
更新日期:2002-06-01 00:00:00
abstract::Rab GTPases are post-translationally geranylgeranylated on their C-terminal cysteines by Rab geranylgeranyl transferase (RabGGTase) and this modification is essential for their biological activity. Rab Escort Protein (REP) is a molecular chaperone that assists in the prenylation reaction carried out by RabGGTase. Muta...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(02)00506-5
更新日期:2002-10-01 00:00:00
abstract::The gene that was inferred to encode the TyrR protein of Haemophilus influenzae Rd was synthesized by polymerase chain reaction and inserted into a T7-based expression vector. Methods were developed to overexpress the TyrR protein of H. influenzae in Escherichia coli and to purify the protein on a large scale. Both in...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.1997.0757
更新日期:1997-07-01 00:00:00
abstract::The Plasmodium falciparum cysteine protease falcipain-2 is a potential new target for antimalarial chemotherapy. In order to obtain large quantities of active falcipain-2 for biochemical and structural analysis, a systematic assessment of optimal parameters for the expression and refolding of the protease was carried ...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1416
更新日期:2001-06-01 00:00:00
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
更新日期:1999-11-01 00:00:00
abstract::Separation of copurifying protease activity from recombinant human Müllerian inhibiting substance (rhMIS) bound to a monoclonal antibody immunoaffinity column by a high-salt wash results in cleaner preparations of rhMIS resistant to cleavage upon storage. In addition, an inhibitor of rhMIS antiproliferative activity i...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/1046-5928(92)90020-w
更新日期:1992-06-01 00:00:00
abstract::The purpose of this work was to characterize the functions of two sesquiterpene synthase genes from moso bamboo (Phyllostachys edulis). Two novel sesquiterpene synthase genes, belonging to the Tpsa subfamily, were isolated from moso bamboo. MoTPS2 was 1641 bp in length and encoded a protein of 63 kDa, whereas MoTPS6 w...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2015.11.019
更新日期:2016-04-01 00:00:00
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
更新日期:1997-04-01 00:00:00
abstract::One of the self-protection mechanisms in Pseudomonas syringae pv. tabaci, a pathogen of tobacco wildfire, is thought to be due to its tabtoxin-resistance gene (ttr). In this study, the ttr gene was inserted into an expression vector, pQE30, and successfully expressed in Escherichia coli M15 at high levels. The purifie...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2001.1586
更新日期:2002-04-01 00:00:00
abstract::A mammalian expression vector with features optimized for simple expression and purification of secreted proteins has been developed. This vector was constructed to facilitate X-ray crystallographic studies of cysteine-rich glycoproteins that are difficult to express by other means. Proteins expressed with this vector...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1006/prep.2000.1331
更新日期:2000-12-01 00:00:00
abstract::The limit dextrinase inhibitor (LDI) from barley seeds acts specifically on limit dextrinase (LD), an endogenous starch debranching enzyme. LDI is a 14 kDa hydrophobic protein containing four disulfide bonds and one unpaired thiol group previously found to be either glutathionylated or cysteinylated. It is a member of...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2011.04.009
更新日期:2011-10-01 00:00:00
abstract::Mitochondrial 2,4-dienoyl-CoA reductase is a key enzyme for the beta-oxidation of unsaturated fatty acids. The cDNA of the full-length human mitochondrial 2,4-dienoyl-CoA reductase was previously cloned as pUC18::DECR. PCR methodologies were used to subclone the genes encoding various truncated human mitochondrial 2,4...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/s1046-5928(03)00191-8
更新日期:2003-10-01 00:00:00
abstract::A full-length cDNA sequence of plant type CRY (designated Hae-P-CRY) was cloned from the green alga Haematococcus pluvialis. The cDNA sequence was 3608 base pairs (bp) in length, which contained a 2988-bp open reading frame encoding 995 amino acids with molecular mass of 107.7 kDa and isoelectric point of 6.19. Multip...
journal_title:Protein expression and purification
pub_type: 杂志文章
doi:10.1016/j.pep.2020.105633
更新日期:2020-08-01 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