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Protein Expr Purif 2019 Apr 17. Epub 2019 Apr 17.

Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan, Taiwan; Department of Pharmacology, College of Medicine, National Cheng Kung University, Tainan, Taiwan. Electronic address:

Survivin is a well-known inhibitor-of-apoptosis proteins family member and a promising molecular target for anti-cancer treatment. However, it is widely accepted that survivin is only a "semi-druggable" target and development of survivin-specific small molecule inhibitors has shown to be difficult. In this study, we demonstrated that a histidine-tagged survivin T34A-C84A mutated protein (T34A-C84A-dNSur-His) can be produced using a bacterial recombinant protein expression system [E. Read More

Protein Expr Purif 2019 Apr 15;160:66-72. Epub 2019 Apr 15.

Department of Pharmaceutical Biotechnology, Isfahan Pharmaceutical Research Center, Faculty of Pharmacy, Isfahan University of Medical Sciences, Isfahan, Iran. Electronic address:

The most common approaches to improve soluble expression of heterologous proteins are applications of molecular chaperones such as DnaK, DnaJ, GrpE, GroEL and GroES. The aim of present study was to enhance soluble expression of granulocyte-macrophage colony-stimulating factor (GM-CSF) in Escherichia coli by different approaches including modification of cultivation and induction conditions, and thermally, genetically and chemically enhancement of expression of cellular chaperones. To genetically enhance amount of molecular chaperones, co-expression of pET28-GM-CSF and pKJE7 plasmids was performed. Read More

Protein Expr Purif 2019 Apr 10;160:56-65. Epub 2019 Apr 10.

PATH's Malaria Vaccine Initiative (MVI), 455 Massachusetts Avenue NW, Suite 1000, Washington, DC, 20001-2621, USA.

In an effort to control and eventually eliminate malaria, the development of transmission-blocking vaccines has long been sought. However, few antigens have been evaluated in clinical trials, often due to limitations in the expression and purification of the antigen in sufficient yield and quality. Pfs230, a surface antigen of gametocytes, has recently advanced to clinical evaluation as a conjugate vaccine using the Pseudomonas aeruginosa exoprotein A carrier protein. Read More

Protein Expr Purif 2019 Apr 4;160:45-55. Epub 2019 Apr 4.

Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada, M5S 3G5; Department of Chemical and Physical Sciences, University of Toronto Mississauga, Mississauga, ON, Canada, L5L 1C6; Department of Chemistry, University of Toronto, Toronto, Ontario, M5S 3H6, Canada. Electronic address:

Packaging the viral genome in the head of double-stranded DNA viruses, such as bacteriophages, requires the activity of a terminase. The bacteriophage terminase consists of a small terminase subunit (TerS), which binds the viral DNA, and a large terminase subunit (TerL) that possesses the ATPase and nuclease activities for packaging the DNA in the phage head. Some phages require additional components for DNA packaging, such as the HNH endonuclease gp74 in the bacteriophage HK97. Read More

Protein Expr Purif 2019 Apr 3. Epub 2019 Apr 3.

Institute for Organic Chemistry and Chemical Biology, Center of Biomolecular Magnetic Resonance, Goethe University Frankfurt, Max-von-Laue-Straße 7, 60438, Frankfurt Am Main, Germany.

Optimization of coding sequences to maximize protein expression yield is often outsourced to external service providers during oligonucleotide synthesis and thus remains a black box for many researchers. The presented software tool CodonWizard offers scientists a free, intuitive, easy-to-use software program for customized codon optimization. It provides features for analysis and completely customized modification/optimization of codon usage of any given input sequence data (protein/RNA/DNA) using combinatory algorithms. Read More

Protein Expr Purif 2019 Mar 31;160:28-35. Epub 2019 Mar 31.

Departamento de Química, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, 14040-901, Ribeirão Preto, SP, Brazil. Electronic address:

Nanoparticles can act as support materials for enzymatic immobilization, introducing a balance of characteristics that modulate the efficiency of biocatalysts, such as specific surface area, resistance to mass transfer and effective enzymatic loading. Magnetic nanoparticles can be easily separated using an external magnetic field, and in this work two recombinant enzymes, the β-glucosidase from Humicola insolens (Bglhi) and the endoglucanase from Scytalidium thermophilum (Egst) were immobilized on synthetized FeO nanoparticles derivatized with chitosan/glutaraldehyde/N-(5-amino-1-carboxy-pentyl) iminodiacetic acid and functionalized with NiCl. The immobilization yields were about 20% for Bglhi and Egst with efficiencies of 132% and 115%, respectively. Read More

Protein Expr Purif 2019 Mar 26. Epub 2019 Mar 26.

College of Food and Biological Engineering, Jimei University, Xiamen, 361021, China; Fujian Provincial Key Laboratory of Food Microbiology and Enzyme Engineering, Xiamen, 361021, China; Research Center of Food Biotechnology of Xiamen City, Xiamen, 361021, China; Key Laboratory of Systemic Utilization and In-depth Processing of Economic Seaweed, Xiamen Southern Ocean Technology Center of China, Xiamen, 361021, China. Electronic address:

A new alginate lyase gene of algl17 was cloned from an alginate-degrading marine bacterium Microbulbifer sp. ALW1. The gene contained 2220 bp and encoded a 739-amino acid protein classified into the PL-17 family. Read More

Protein Expr Purif 2019 Mar 27;160:36-44. Epub 2019 Mar 27.

Department of Biotechnology, Himachal Pradesh University, Summer Hill, Shimla, Himachal Pradesh, 171005, India. Electronic address:

Xanthine oxidase (EC 1.17.3. Read More

Protein Expr Purif 2019 Mar 26;160:7-10. Epub 2019 Mar 26.

Department of Physiology, Graduate School of Medical & Dental Sciences, Kagoshima University, Kagoshima, 890-8544, Japan. Electronic address:

Both recombinant glutathione-S-transferase (GST)-fused and GST-cleaved fragments of an L-type voltage-gated Ca channel (Cav1.2) are used frequently in GST pull-down assays to investigate the interactions between regulatory proteins and the Cav1.2 channel. Read More

Protein Expr Purif 2019 Mar 25;160:1-6. Epub 2019 Mar 25.

Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand; Biofuels by Biocatalysts Research Unit, Department of Botany, Faculty of Science, Chulalongkorn University, Bangkok, 10330, Thailand. Electronic address:

Filamentous fungi are the most common industrial xylanase producers. In this study, the xynA gene encoding xylanase A of Penicilium citrinum was successfully synthesized and expressed in Yarrowia lipolytica under the control of the strong constitutive TEF promoter. Native and preproLIP2 secretion signals were used for comparison of the expression and secretion level. Read More

Protein Expr Purif 2019 Jul 25;159:75-82. Epub 2019 Mar 25.

CSL Limited, BIO21 Institute, 30 Flemington Road, Parkville, Victoria, 3010, Australia; CSL Behring GmbH, R&D, Emil-von-Behring Str. 76, Marburg, 35041, Germany. Electronic address:

The ability to engineer monoclonal antibodies (mAbs) with high specificity made mAbs the fastest growing segment in the drug market. mAbs represent 8 of the top 20 selling drugs with combined sales of more than 57 billion US$ per year. The ability to purify large numbers of mAbs with sufficient yields for initial screening campaigns has direct impact on the timelines of a project. Read More

Protein Expr Purif 2019 Jul 24;159:69-74. Epub 2019 Mar 24.

Laboratory of Creative Science for Insect Industries, Kyushu University Graduate School of Bioresource and Bioenvironmental Sciences, Motooka 744, Nishi-ku, Fukuoka, 819-0395, Japan. Electronic address:

Human granulocyte-macrophage colony stimulating factor (hGM-CSF) is a hematopoietic growth factor. It is widely employed as a therapeutic agent targeting neutropenia in cancer patients undergoing chemotherapy and in patients with AIDS or after bone marrow transplantation. In this study, we constructed the recombinant baculoviruses for the expression of recombinant hGM-CSF (rhGM-CSF) with two small affinity tags (His-tag and Strep-tag) at the N or C-terminus. Read More

Protein Expr Purif 2019 Jul 21;159:49-52. Epub 2019 Mar 21.

Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, Av Universidad 1001, Colonia Chamilpa, Cuernavaca, 62209, Morelos, Mexico.

Metagenomic libraries are a novel and powerful approach to seek for pathways involved in xenobiotic degradation, since this technique abolishes the need for cultivating microorganisms that otherwise would be overlooked if they cannot grow on standard laboratory media and conditions. In this paper, we describe the expression, purification and characterization of a novel metagenomic thioesterase which was described to be involved in phenylacetic acid degradation (A. Sánchez-Reyes, R. Read More

Protein Expr Purif 2019 Jul 22;159:53-59. Epub 2019 Mar 22.

Department of Inorganic and Analytical Chemistry, University of Szeged, Dóm tér 7, H-6720, Szeged, Hungary. Electronic address:

The role of the termini of protein sequences is often perturbed by remnant amino acids after the specific protease cleavage of the affinity tags and/or by the amino acids encoded by the plasmid at/around the restriction enzyme sites used to insert the genes. Here we describe a method for affinity purification of a metallonuclease with its precisely determined native termini. First, the gene encoding the target protein is inserted into a newly designed cloning site, which contains two self-eliminating BsmBI restriction enzyme sites. Read More

Protein Expr Purif 2019 Mar 21;160:19-27. Epub 2019 Mar 21.

State Key Laboratory of Food Nutrition and Safety, China International Scientific & Technological Cooperation Base for Health Biotechnology, College of Food Engineering and Biotechnology, Tianjin University of Science & Technology, Tianjin, 300457, People's Republic of China; Obesita & Algaegen LLC, College Station, TX, 77845, USA. Electronic address:

Hispidalin is a novel antimicrobial peptide isolated from the seeds of Benincasa hispida and is reported to have broad antimicrobial activity against various bacterial and fungal pathogens. To produce significant amounts of Hispidalin, a recombinant Hispidalin with an N-terminal 6 × His tag and an enterokinase sequence, for the first time, was successfully expressed in Escherichia coli or Pichia pastoris cell factory. Results showed that the E. Read More

Protein Expr Purif 2019 Jul 17;159:42-48. Epub 2019 Mar 17.

School of Biotechnology and Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University, Wuxi, 214122, China; State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, 214122, China.

Pullulanase is widely used in the starch processing industry as a debranching enzyme. However, extracellular production of pullulanase from recombinant Bacillus subtilis is limited and the loss of plasmids during fermentation of B. subtilis recombinants seriously affects the expression of the foreign protein, especially in large-scale production. Read More

Protein Expr Purif 2019 Jul 14;159:34-41. Epub 2019 Mar 14.

Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, 03722, Republic of Korea; Institute of Life Science and Biotechnology, Yonsei University, Seoul, 03722, Republic of Korea. Electronic address:

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 Fab region of the monoclonal antibody produced in the plant; thus, it is necessary to compare the antigen affinity of this antibody with that of the prototype. In this study, ofatumumab, a fully human anti-CD20 IgG1κ monoclonal antibody used for its non-cross resistance to rituximab, was expressed in Nicotiana benthamiana, and its affinities and efficacies were compared with those of native ofatumumab produced from CHO cells. Read More

Protein Expr Purif 2019 Mar 13;160:11-18. Epub 2019 Mar 13.

Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria, Australia. Electronic address:

Bacteria contain sigma (σ) factors that control gene expression in response to various environmental stimuli. The alternative sigma factors σ and σ bind specifically to the antisigma factor FpvR. These proteins are an essential component of the pyoverdine-based system for iron uptake in Pseudomonas aeruginosa. Read More

Protein Expr Purif 2019 Jul 11;159:83-90. Epub 2019 Mar 11.

Food Safety and Agricultural Research Center, Akdeniz University, Antalya, Turkey; Food Engineering Department, Faculty of Engineering, Akdeniz University, Antalya, Turkey.

Microbial pro-transglutaminase (pro-MTGase) from Streptomyces mobaraensis was expressed in Pichia pastoris (Komagataella phaffii) under the control of constitutive GAP promoter. The single copy of the gene containing clone was grown in shake flasks to determine the optimum conditions for the production of recombinant pro-MTGase. Three temperature (20 °C, 25 °C, 28 °C) and four pH (5, 6, 7, 7. Read More

Protein Expr Purif 2019 Jul 11;159:27-33. Epub 2019 Mar 11.

Department of Biology and Chemistry, National University of Defense Technology, Changsha, 410000, Hunan, China. Electronic address:

Olfactory receptors (OR), a group of classic membrane proteins, plays a vital role in insect reproduction and acclimatization. Deciphering the molecular mechanism of insect olfaction could enhance pest control and environmental protection. Studies on ORs have faced a major bottleneck due to the notoriously strong hydrophobicity of ORs, which results in difficult expression in heterologous cell systems. Read More

Protein Expr Purif 2019 Jul 6;159:60-68. Epub 2019 Mar 6.

Laboratory of Cell Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA. Electronic address:

Human P-glycoprotein (P-gp) is an ATP-binding cassette transporter that has been implicated in altering the pharmacokinetics of anticancer drugs in normal tissues and development of multidrug resistance in tumor cells via drug efflux. There is still no definitive explanation of the mechanism by which P-gp effluxes drugs. One of the challenges of large-scale purification of membrane transporters is the selection of a suitable detergent for its optimal extraction from cell membranes. Read More

Protein Expr Purif 2019 Jul 2;159:21-26. Epub 2019 Mar 2.

Institute of Drug Technology, Oswaldo Cruz Foundation, Rio de Janeiro, RJ, Brazil. Electronic address:

l-asparaginase catalyzes the conversion of l-asparagine to l-aspartate and ammonium. This protein is an important therapeutic enzyme used for the treatment of acute lymphoblastic leukemia. In this study, the asparaginase II-encoding gene ASP3 from Saccharomyces cerevisiae was cloned into the expression vector pET28a in-fusion with a 6x histidine tag and was expressed in Escherichia coli BL21 (DE3) cells. Read More

Protein Expr Purif 2019 Jun 1;158:65-73. Epub 2019 Mar 1.

Technology and Process Development (TPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China. Electronic address:

Protein A chromatography is generally less effective at removing antibody aggregates under typical conditions. We recently developed a method that can significantly improve Protein A's aggregate removal capability. This method involves adding calcium chloride/polyethylene glycol (PEG) or sodium chloride/PEG combination to wash and elution buffers. Read More

Protein Expr Purif 2019 Jul 1;159:1-9. Epub 2019 Mar 1.

Division of Biochemistry, CSIR-Central Drug Research Institute, Lucknow, 226031, Uttar Pradesh, India.

Arginase catalyzes the first committed step in the biosynthesis of polyamines that enable cell growth and hence potential drug target for the treatment of leishmaniasis. The arginase from Leishmania donovani (LdARG) was cloned, overexpressed and characterized. Analysis of the deduced amino acid sequence of LdARG with homologous enzyme from other trypanosomatids arginases identified a non-conserved 12 residues long segment VWGLIERTFLSA from position 161-172. Read More

Protein Expr Purif 2019 Jun 1;158:74-80. Epub 2019 Mar 1.

Collage of Life Sciences, Nankai University, Tianjin, 300071, China. Electronic address:

Receptor tyrosine kinase like orphan receptor 2 (ROR2) is a co-receptor for some Wnt proteins including Wnt5a that activate the noncanonical Wnt/planar cell polarity (PCP) signaling pathway. Upregulation of ROR2 is associated with several cancer forms. The extracellular region of ROR2, which contains an immunoglobulin (Ig)-like domain, a Frizzled like cysteine-rich domain (CRD) and a Kringle domain, is a potential anticancer drug target. Read More

Protein Expr Purif 2019 Jun 28;158:59-64. Epub 2019 Feb 28.

Technology and Process Development (TPD), WuXi Biologics, 288 Fute Zhong Road, Waigaoqiao Free Trade Zone, Shanghai, 200131, China. Electronic address:

Post-load column wash in Protein A chromatography can effectively improve host cell protein (HCP) clearance. A commonly used wash additive for this purpose is sodium chloride. However, the adequate amount of sodium chloride required for effective HCP clearance is less consistent in literature. Read More

Protein Expr Purif 2019 Jun 27;158:81-88. Epub 2019 Feb 27.

Department of Molecular Biology, Chonbuk National University, 664-14 Dukjindong, Jeonju, Jeollabuk-do, 54896, Republic of Korea; National Institute of Horticultural & Herbal Science (NIHHS), Rural Development Administration (RDA), Wanju, Jeollabuk-do, 55365, Republic of Korea. Electronic address:

Gaucher disease is an inherited metabolic disease caused by genetic acid β -glucosidase (GBA) deficiency and is currently treated by enzyme replacement therapy. For uptake into macrophages, GBA needs to carry terminal mannose residues on their N-glycans. Knockout mutant rice of N-acetylglucosaminyltransferase-I (gnt1) have a disrupted N-glycan processing pathway and produce only glycoproteins with high mannose residues. Read More

Protein Expr Purif 2019 Jun 23;158:36-43. Epub 2019 Feb 23.

School of Food Science and Pharmaceutical Engineering, Nanjing Normal University, Nanjing, 210046, China. Electronic address:

Insect-derived serine protease inhibitors (serpins) exhibit multiple inhibitory activities. Todate some functional roles for serpins in Hyphantria cunea Drury have been identified. Here, new functional features of the H. Read More

Protein Expr Purif 2019 Jul 23;159:10-16. Epub 2019 Feb 23.

Key Laboratory for Food Microbial Technology of Zhejiang Province, Department of Biotechnology, Zhejiang Gongshang University, Hangzhou, Zhejiang, 310018, People's Republic of China. Electronic address:

Plantaricin JK (PlnJK) is a Class IIb LAB bacteriocin that includes two peptides; i.e., PlnJ and PlnK, which can synergistically halt many types of gram-positive bacteria, including food spoilage organisms. Read More

Protein Expr Purif 2019 Jul 18;159:17-20. Epub 2019 Feb 18.

Dept. of Solid-State NMR, Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2300RA, Leiden, the Netherlands. Electronic address:

Photosystem II subunit S (PsbS) is a membrane protein that plays an exclusive role in non-photochemical quenching for photoprotection of plants under high-light conditions. The activation mechanism of PsbS and its pH-induced conformational changes are currently unknown. For structural investigation of PsbS, effective synthesis of PsbS with selective isotope or electron-spin labels or non-natural amino acids incorporated would be a great asset. Read More

Protein Expr Purif 2019 Jun 15;158:27-35. Epub 2019 Feb 15.

School of Molecular and Cell Biology, University of Witwatersrand, Private Bag 3, Wits, 2050, Johannesburg, South Africa; Department of Biotechnology and Food Technology, University of Johannesburg, P.O. Box 17011, Doornfontein, 2028, Johannesburg, South Africa.

PbrD is a lead (II) binding protein encoded by the pbr lead resistance operon found exclusively in Cupriavidus metallidurans CH34. Its ability to sequester Pb(II) shows potential for it to be developed as a biosorbent for Pb in the bioremediation of contaminated wastewaters. In this study the pbrD gene from C. Read More

Protein Expr Purif 2019 Jun 14;158:44-50. Epub 2019 Feb 14.

Facultad de Farmacia y Bioquímica, Cátedra de Biotecnología, Universidad de Buenos Aires, Junín 956, 1113, Buenos Aires, Argentina; Instituto de Nanobiotecnología (NANOBIOTEC), CONICET-Universidad de Buenos Aires, Junín 956, 1113, Buenos Aires, Argentina. Electronic address:

Expression of recombinant proteins with baculovirus-infected insect larvae is a scarcely investigated alternative in comparison to that in insect cell lines, a system with growing popularity in the field of biotechnology. The aim of this study was to investigate the chromatographic behavior and physicochemical properties of the proteome of Rachiplusia nu larvae infected with recombinant Autographa californica multiple nucleopolyhedrosis virus (AcMNPV), in order to design rational purification strategies for the expression of heterologous proteins in this very complex and little-known system, based on the differential absorption between target recombinant proteins and the system's contaminating ones. Two-dimensional (2D) gel electrophoresis showed differences in the protein patterns of infected and non-infected larvae. Read More

Protein Expr Purif 2019 Jun 10;158:1-8. Epub 2019 Feb 10.

School of Agriculture, Ludong University, Yantai, 264025, China.

Grass carp reovirus (GCRV) is one of the most serious pathogens threatening grass carp (Ctenopharyngodon idellus) production and results in high mortality in China. VP7 from GCRV is involved in viral infection and could be suitable for developing vaccines for the control of GCRV infection. To obtain a genetically engineered vaccine and a plant-based oral vaccine and to evaluate their immune efficacy as an oral vaccine against GCRV, cholera toxin B subunit (CTB) of Vibrio cholerae fused to VP7 (CTB-VP7) was transformed into BL21(DE3) for expression. Read More

Protein Expr Purif 2019 Jun 8;158:15-19. Epub 2019 Feb 8.

Department of Integrated Structural Biology, Institute of Genetics and Molecular and Cellular Biology, CNRS UMR710, INSERM U964, University of Strasbourg, Strasbourg, 67000, France. Electronic address:

Human elongation factor 2 is the translocase that is responsible for the movement of tRNA from the A- to P- and P- to E-site on the ribosome during the elongation phase of translation. Being a vital factor of protein biosynthesis, its function is highly controlled and regulated. It has been implicated in numerous diseases and pathologies, and as such it is important to have a source for isolated pure and active protein for biomedical and biochemical studies. Read More

Protein Expr Purif 2019 Jun 7;158:9-14. Epub 2019 Feb 7.

Gustaf H. Carlson School of Chemistry & Biochemistry, Clark University, 950 Main St, Worcester, MA, 01610, USA. Electronic address:

Homeodomain transcription factors (HD TFs) are a large class of evolutionarily conserved DNA binding proteins that contain a basic 60-amino acid region required for binding to specific DNA sites. In Drosophila melanogaster, many of these HD TFs are expressed in the early embryo and control transcription of target genes in development through their interaction with cis-regulatory modules. Previous studies where some of the Drosophila HD TFs were purified required the use of strong denaturants (i. Read More

Protein Expr Purif 2019 Jun 6;158:20-26. Epub 2019 Feb 6.

Department of Chemistry, University of Umeå, SE-901 87, Umeå, Sweden. Electronic address:

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. Read More

Protein Expr Purif 2019 May 6;157:86-91. Epub 2019 Feb 6.

Department of Chemistry, University Konstanz, DE-78464 Konstanz, Germany; Konstanz Research School Chemical Biology (KoRS-CB), University Konstanz, DE-78464 Konstanz, Germany. Electronic address:

High resolution NMR spectroscopy is a seminal method in modern structural biology to obtain insights into proteins' structure, dynamics and function at dilute condition as well as in a cell-like environment or even intracellularly. Usually, H, N or C nuclei are predominantly used for the characterization of the protein of interest. These measurements are limited due to the wealth of chemical shifts and background signals arising from all molecules present in the NMR test tube. Read More

Protein Expr Purif 2019 May 5;157:63-69. Epub 2019 Feb 5.

Institute of Biotechnology and Antibiotics, Starościńska 5, Warszawa, 02-516, Poland.

The number of people with diabetes is estimated to be over 370 million, in 2030 it will increase to 552 million. In Poland, the number of people with diabetes is estimated to be 3.5 million (9. Read More

Protein Expr Purif 2019 May 5;157:57-62. Epub 2019 Feb 5.

Guangdong Provincial Key Laboratory for Biotechnology Candidate Drug Research, School of Biosciences and Biopharmaceutics, Guangdong Pharmaceutical University, Guangzhou, 510006, PR China. Electronic address:

Epidermal growth factor receptor (EGFR) is an effective target for the treatment of many epithelial cancers. However, EGFR inhibitors have low clinical response rates and are prone to drug resistance arising from mutations and heterodimerization of EGFR. Therefore, targeting the highly conserved dimer interface of EGFR may be an effective strategy for improving the clinical response of anti-EGFR therapies. Read More

Protein Expr Purif 2019 May 1;157:50-56. Epub 2019 Feb 1.

Shenzhen Key Laboratory of Fermentation, Purification and Analysis, Shenzhen Polytechnic, Shenzhen, 518055, China. Electronic address:

The development of antibiotic-resistant bacteria has become a major public health problem, prompting the search for alternative solutions. Tachyplesin I (TP-I) is an antimicrobial peptide, which exhibits potent and broad-spectrum activities against bacteria, fungi, viruses, and tumor cells. However, limited amounts of TP-I produced in horseshoe crab restrict its large-scale use. Read More

Protein Expr Purif 2019 May 30;157:28-35. Epub 2019 Jan 30.

Belozersky Institute of Physico-Chemical Biology, Lomonosov Moscow State University, Lenin's Hills 1 Bldg.40, Moscow, 119234, Russian Federation.

The goal of the present work was to produce glyceraldehyde-3-phospate dehydrogenase from M. tuberculosis in E. coli cells in soluble and catalytically active form and to elaborate a method for the purification of the recombinant enzyme. Read More

Protein Expr Purif 2019 May 29;157:42-49. Epub 2019 Jan 29.

Medical Biotechnology Department, Biotechnology Research Center, Pasteur Institute of Iran, Tehran, Iran. Electronic address:

The secretory production of heterologous proteins in E. coli has revolutionized biotechnology. Efficient periplasmic production of foreign proteins in E. Read More

Protein Expr Purif 2019 May 30;157:70-85. Epub 2019 Jan 30.

Department of Chemistry, Trent University, 1600 West Bank Drive, Peterborough, ON, K9J 0G2, Canada. Electronic address:

NMR is an important method in the structural and functional characterization of proteins, but such experiments typically require isotopic labelling because of the low natural abundance of the nuclei of interest. Isotope-labelled protein for NMR experiments is typically obtained from IPTG-inducible bacterial expression systems in a minimal media that contains labelled carbon or nitrogen sources. Optimization of expression conditions is crucial yet challenging; large amounts of labelled protein are desired, yet protein yields are lower in minimal media, while the labelled precursors are expensive. Read More

Protein Expr Purif 2019 May 29;157:21-27. Epub 2019 Jan 29.

Department of Chemistry and Biochemistry, Faculty of Chemistry and Chemical technology, University of Ljubljana, Večna pot 113, 1000, Ljubljana, Slovenia. Electronic address:

Cathepsin C is a tetrameric lysosomal protease that acts as a dipeptidyl-peptidase due to the presence of the exclusion domain that is unique among papain-like cysteine proteases. Here we describe a recombinant form of cathepsin C lacking its exclusion domain (CatCΔEx) produced in a bacterial expression system (E. coli). Read More

Protein Expr Purif 2019 May 26;157:17-20. Epub 2019 Jan 26.

State Key Laboratory of Biocatalysis and Enzyme Engineering, School of Life Sciences, Hubei University, China; Hubei Key Laboratory of Industrial Biotechnology, School of Life Sciences, Hubei University, China; Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, School of Life Sciences, Hubei University, China. Electronic address:

Human lipopolysaccharide (LPS) binding protein (LBP) is a ∼60 kDa glycosylated protein that mediates potent innate immune against invading Gram-negative bacteria by recognition of LPS in their outer membranes. To date, there is no method for efficient production of bioactive LPS-free LBP at sufficient amounts through prokaryotic expression system. Here we present a simple approach for rapid preparation of human LBP from a LPS-eliminated E. Read More

Protein Expr Purif 2019 May 14;157:9-16. Epub 2019 Jan 14.

Department of Biotechnology, Dayananda Sagar College of Engineering, Kumaraswamy Layout, Shavige Malleswara Hills, Bengaluru, 78, Karnataka, India.

A putrescine monooxygenase from Shewanella putrefaciens 95 (SpPMO) is the initial enzyme catalyzing the hydroxylation of putrescine to N-hydroxyl putrescine, the precursor for the synthesis of a siderophore putrebactin was identified. This PMO clustered together with known characterized NMOs from Shewanella baltica, Bordetella pertussis, Erwinia amylovora, Streptomyces sp. Gordonia rubripertincta, Pseudomonas aeruginosa and outgrouped from Escherichia coli, Nocardia farcinica, and Rhodococcus erythropolis. Read More

Protein Expr Purif 2019 May 9;157:36-41. Epub 2019 Jan 9.

Division of Biotechnology, Faculty of Agro-Industry, Chiang Mai University, Chiang Mai, 50100, Thailand; Research Center for Multidisciplinary Approaches to Miang, Chiang Mai University, Chiang Mai, 50200, Thailand. Electronic address:

Lactobacillus pentosus BA-7 and L. pentosus QA1-5 are tannin-tolerant lactic acid bacteria that were isolated from Miang, a traditional fermented tea-leaf found in northern Thailand and a tannin-rich substrate. Tannase encoding genes were isolated, cloned and overexpressed in Escherichia coli BL21(DE3). Read More

Protein Expr Purif 2019 Apr 7;156:66-71. Epub 2019 Jan 7.

State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, PR China. Electronic address:

Naturally-occurring orange carotenoid protein (OCP) is synthesized in cyanobacteria and red algae for photoprotection. Holo-OCP can be produced with three plasmids in E. coli, which needs two inducers (arabinose and isopropyl β-D-thiogalactoside) to initiate two processes: one for generation of carotenoid and the other for generation of apo-OCP, so takes about two days. Read More

Protein Expr Purif 2019 Apr 7;156:58-65. Epub 2019 Jan 7.

Faculty of Agriculture, Kagawa University, 2393 Ikenobe, Miki-cho, Kita-gun, Kagawa, 761-0795, Japan. Electronic address:

The novel isolated Rhizobium sp. S10 was identified as d-glucoside 3-dehydrogenase (G3DH) producing microbe. Therefore, the gene encoding for G3DH from Rhizobium sp. Read More

Protein Expr Purif 2019 Apr 4;156:50-57. Epub 2019 Jan 4.

Applied and Molecular Microbiology, Faculty of Agriculture, Kagoshima University, 1-21-24 Korimoto, Kagoshima, 890-0065, Japan.

Insoluble expression of intrinsically soluble proteins with native activity is potentially a promising alternative to soluble expression of folded protein or insoluble expression of unfolded protein requiring refolding. Here, we attempted to express highly soluble halophilic His-rich metal binding protein (HP) as insoluble inclusion bodies with native metal-binding activity using insolubilizing nona-peptide (Ins), GILQINSRW, derived from hen egg white lysozyme (His-InsHP). About 80% of expressed His-InsHP was localized in inclusion bodies in Na-phosphate/NaCl buffer, pH 7. Read More