Generation and identification of variants with improved purification yield of Bowman-Birk protease inhibitors carrying protein binding loops.

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

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

更新日期：1996-12-01 00:00:00

abstract：:Haloalkane dehalogenase from Xanthobacter autotrophicus was efficiently expressed in Escherichia coli BL21 (DE3) and E. coli JM101. After introduction of restriction sites by PCR the haloalkane dehalogenase gene (dhlA) was translationally fused behind the T7 (phi 10), trc, and tac promoters. This resulted in expressio...

journal_title：Protein expression and purification

authors： Schanstra JP,Rink R,Pries F,Janssen DB

更新日期：1993-10-01 00:00:00

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

authors： Li P,Gu Q,Wu X

更新日期：2016-10-01 00:00:00

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...

journal_title：Protein expression and purification

authors： Lee SG,Hong SP,Choi YH,Chung YJ,Sung MH

更新日期：1997-12-01 00:00:00

abstract：:The market of therapeutic glycoproteins (including coagulation factors, antibodies, cytokines and hormones) is one of the profitable, fast-growing and challenging sectors of the biopharmaceutical industry. Although mammalian cell culture is still expensive and technically complex, the ability to produce desired post-t...

journal_title：Protein expression and purification

authors： Rosenlöcher J,Weilandt C,Sandig G,Reinke SO,Blanchard V,Hinderlich S

更新日期：2015-11-01 00:00:00

abstract：:PagP, a beta-barrel membrane protein found in Gram-negative bacteria, expresses robustly in inclusion bodies when its signal sequence is removed. We have developed a new fusion protein expression system based on PagP and demonstrated its utility in the expression of the unstructured N-terminal region of human cardiac ...

journal_title：Protein expression and purification

authors： Hwang PM,Pan JS,Sykes BD

更新日期：2012-09-01 00:00:00

abstract：:To investigate the expression and purification of an unstable heterologous protein in Pichia pastoris, the cDNA of H5-lysozyme, a hen egg lysozyme mutant with a hydrophobic pentapeptide (Phe-Phe-Val-Ala-Pro) fused to the carboxyl terminus, was integrated into the genome of P. pastoris. It was found that medium composi...

journal_title：Protein expression and purification

authors： Liu ST,Saito A,Azakami H,Kato A

更新日期：2003-02-01 00:00:00

abstract：:Refolding of proteins from inclusion bodies is affected by several factors, including solubilization of inclusion bodies by denaturants, removal of the denaturant, and assistance of refolding by small molecule additives. We will review key parameters associated with (1) conformation of the protein solubilized from inc...

journal_title：Protein expression and purification

authors： Tsumoto K,Ejima D,Kumagai I,Arakawa T

更新日期：2003-03-01 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

authors： Ribó M,delCardayré SB,Raines RT,de Llorens R,Cuchillo CM

更新日期：1996-05-01 00:00:00

abstract：:The promyelocytic leukemia (PML) gene is involved in the 15/17 chromosomal translocation of acute promyelocytic leukemia (APL). It encodes a nuclear phosphoprotein containing an alpha-helical coiled-coil domain with four heptad repeats. The heptad repeats consist of four clusters of hydrophobic amino acids that mediat...

journal_title：Protein expression and purification

authors： Antolini F,Lo Bello M,Sette M

更新日期：2003-05-01 00:00:00

abstract：:Accurate diagnosis is essential for the treatment, prevention, and control of tuberculosis. Poor specificity of the tuberculin skin test in BCG-vaccinated populations and constraints to implementation of PCR and CMI-based diagnostic assays in developing countries warrant development of easy-to perform robust serologic...

journal_title：Protein expression and purification

authors： Kulshrestha A,Gupta A,Verma N,Sharma SK,Tyagi AK,Chaudhary VK

更新日期：2005-11-01 00:00:00

abstract：:Most parasitic protozoa lack the de novo purine biosynthetic pathway and rely exclusively on the salvage pathway for their purine nucleotide requirements. Enzymes of the salvage pathway are, therefore, candidate drug targets. We have cloned the Plasmodium falciparum adenylosuccinate synthetase gene. In the parasite, a...

journal_title：Protein expression and purification

authors： Jayalakshmi R,Sumathy K,Balaram H

更新日期：2002-06-01 00:00:00

abstract：:The major active form of human thrombin, alpha-thrombin, was analyzed by hydrophobic interaction high-performance liquid chromatography (HIC-HPLC). The chromatographic system included a polymeric phenyl column and elution was performed by a gradient, 2-0M sodium chloride (5-20 min). Total analysis time was 30 min per ...

journal_title：Protein expression and purification

authors： Karlsson G

更新日期：2003-01-01 00:00:00

abstract：:Recombinant expression of human membrane proteins in large quantities remains a major challenge. Expression host is an important variable to screen for high-level production of membrane proteins. Using the green fluorescent protein (GFP) as a reporter, we screened the expression of a human multi-pass membrane protein ...

journal_title：Protein expression and purification

authors： Cai H,Yao H,Li T,Tang Y,Li D

更新日期：2019-12-01 00:00:00

abstract：:Plants produce a large number of cellulases that are either secreted or anchored in the plasma membrane where they likely function in various aspects of cellulose synthesis, modification and degradation during plant growth and development. Very few of these enzymes have been characterized in any detail, however. Here ...

journal_title：Protein expression and purification

authors： Tawde MD,Freimuth P

更新日期：2012-10-01 00:00:00

abstract：:L-Threonine dehydrogenase was purified 10,000-fold to a specific activity approximately 300 mumol.min-1.mg-1 protein from porcine liver mitochondria. Purification to apparent homogeneity was achieved by sequential chromatography on DEAE Sepharose FF, Affi-Gel Blue, Sephacryl S-200, Matrex Gel Red A, and Matrex Gel Gre...

journal_title：Protein expression and purification

authors： Kao YC,Davis L

更新日期：1994-10-01 00:00:00

abstract：:The superoxide dismutases (EC 1.15.1.1) are a family of enzymes that catalyze the dismutation of superoxide radical anion to dioxygen and hydrogen peroxide. The active site contains a critical metal ion such as manganese, iron, or copper. The copper-containing protein also has one zinc ion bound per subunit. The stand...

journal_title：Protein expression and purification

authors： Sutter B,Bounds PL,Koppenol WH

更新日期：2000-06-01 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

authors： Hang W,Gujar A,Zhang H,Xu W,Zhao C,Zhu X,Xue J,Zhang C,Ji C,Qin S,Li R,Cui H

更新日期：2020-08-01 00:00:00

abstract：:Whey acidic proteins (WAP) belong to a large gene family of antibacterial peptides that perform critical immune system functions. The function of human epididymis protein 4 (HE4), a 124-amino acid long polypeptide that has two whey acidic protein four-disulfide core (WFDC) domains, is not well studied. Here, a fusion ...

journal_title：Protein expression and purification

authors： Hua L,Liu Y,Zhen S,Wan D,Cao J,Gao X

更新日期：2014-10-01 00:00:00

abstract：:Mammalian target of rapamycin (TOR) controls cell growth and metabolism in response to the availability of nutrients, growth factors, and the cellular energy status. Misregulation of TOR can result in a pathogenic increase or decrease in organ size and in cancer. TOR can be inhibited by binding of a complex of rapamyc...

journal_title：Protein expression and purification

authors： Dames SA

更新日期：2008-05-01 00:00:00

abstract：:Ciliary neurotrophic factor (CNTF) is a promising candidate for the treatment of neurodegenerative or metabolic diseases, but suffers rapid clearance in body. Herein we constructed a new long-acting recombinant human CNTF (rhCNTF) by genetic fusion with an albumin-binding domain (ABD) through a flexible peptide linker...

journal_title：Protein expression and purification

authors： Xu L,Zhang C,Liu L,Zhang Y,Wang Q,Wang J,Liu Y,Su Z

更新日期：2017-11-01 00:00:00

abstract：:An improved, simplified method for the purification of recombinant, tagged DnaA proteins is described. The presented protocol allowed us to purify soluble DnaA proteins from two different bacterial species: Helicobacter pylori and Streptomyces coelicolor, but it can most likely also be used for the isolation of DnaA p...

journal_title：Protein expression and purification

authors： Zawilak-Pawlik AM,Kois A,Zakrzewska-Czerwinska J

更新日期：2006-07-01 00:00:00

abstract：:Plasmids pQE-60 and pQE-30 containing 6 x His-tag sequence were used for expression of fragments of HCV structural and non-structural proteins in Escherichia coli (E. coli). The following fragments were used: core (1-98 aa), NS3 (202-482 aa), and tetramer of hypervariable region 1 (HVR1) of E2 protein. The constructed...

journal_title：Protein expression and purification

authors： Mihailova M,Fiedler M,Boos M,Petrovskis I,Sominskaya I,Roggendorf M,Viazov S,Pumpens P

更新日期：2006-11-01 00:00:00

abstract：:The H-NS protein is one of the major constituents of the nucleoid structure that has been implicated in the DNA packaging and in the global regulation of gene expression. The study of this transcriptional regulator is an effort to fight Xylella fastidiosa, a citrus pathogen responsible for a range of economically impo...

journal_title：Protein expression and purification

authors： Paula DP,Azzoni AR,Siqueira SF,Catani CF,Rosselli LK,de Souza AP

更新日期：2003-11-01 00:00:00

abstract：:Human β-glucuronidase (GUS; EC 3.2.1.31) is a lysosomal enzyme that catalyzes the hydrolysis of β-d-glucuronic acid residues from the non-reducing termini of glycosaminoglycans. Impairment in GUS function leads to the metabolic disorder mucopolysaccharidosis type VII, also known as Sly syndrome. We produced GUS from a...

journal_title：Protein expression and purification

authors： Fratz-Berilla EJ,Ketcham SA,Parhiz H,Ashraf M,Madhavarao CN

更新日期：2017-12-01 00:00:00

abstract：:17β-hydroxysteroid dehydrogenase type 1 (17β-HSD1) mainly catalyzes the reduction of estrone into estradiol. The enzymatic conversion is a critical step in estradiol accumulation in breast tissue, which is a valuable prognosis index of breast cancer disease. However, the source of 17β-HSD1 for inhibitor design is limi...

journal_title：Protein expression and purification

authors： Chen J,Feng W,Zhao Y

更新日期：2017-09-01 00:00:00

abstract：:Endostatin, a 20kDa C-terminal fragment of collagen XVIII, is a potent anti-angiogenic protein and inhibitor of tumor growth. Recombinant endostatin was prepared from Escherichia coli deposited as insoluble, inactive inclusion bodies. In the present study, we produced soluble and biologically active recombinant human ...

journal_title：Protein expression and purification

authors： Xu HM,Zhang GY,Ji XD,Cao L,Shu L,Hua ZC

更新日期：2005-06-01 00:00:00

abstract：:Fusion proteins expressed in bacteria are often insoluble or inefficiently purified by standard procedures previously reported to work well for the non-fused proteins. We report here a simple but general procedure that can be used to quickly customize and optimize the purification of milligram quantities of most GST f...

journal_title：Protein expression and purification

authors： Mercado-Pimentel ME,Jordan NC,Aisemberg GO

更新日期：2002-11-01 00:00:00

abstract：:Previously, we have shown that the entire extracellular domain of the granulocyte-colony stimulating factor receptor (sG-CSFr) produced in Chinese hamster ovary (CHO) cells forms a stable complex with its ligand G-CSF, at a stoichiometry of 2:2. A truncated receptor molecule consisting of the cytokine receptor homolog...

journal_title：Protein expression and purification

authors： Horan TP,Simonet L,Jacobsen R,Mann M,Haniu M,Wen J,Arakawa T,Kuwamoto M,Martin F

更新日期：1998-10-01 00:00:00

abstract：:Ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit epsilon N-methyltransferase (Protein methylase III, Rubisco LSMT, EC 2.1.1.43) catalyzes methylation of the epsilon-amino group of Lys-14 in the large subunit of Rubisco. In this paper, an affinity purification procedure for pea (Pisum sativum L. cv Laxton'...

journal_title：Protein expression and purification

authors： Wang P,Royer M,Houtz RL

更新日期：1995-08-01 00:00:00