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Protein Expr Purif 2019 Feb 18. 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 Feb 15. 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 Feb 14. 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 Feb 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 Feb 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 Feb 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 Feb 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

Protein Expr Purif 2019 May 4;157:1-8. Epub 2019 Jan 4.

Biochemical Engineering Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology (IIT) Guwahati, Guwahati, 781039, Assam, India. Electronic address:

Reverse micellar extraction (RME), a liquid-liquid based separation is a versatile tool for protein purification. A statistical approach was employed for the purification of recombinant glutaminase free anti-cancerous enzyme viz., l-asparaginase II to evaluate the effects of RME in current study. Read More

Protein Expr Purif 2019 Apr 29;156:17-24. Epub 2018 Dec 29.

National & Local United Engineering Lab of Quality Controlling Technology and Instrumentation for Marine Food, College of Life Science, China Jiliang University, Hangzhou, 310018, China.

The ricexip gene, which encodes the rice xylanase-inhibiting protein (riceXIP), was recombinantly expressed in Pichia pastoris GS115 under the control of AOX1 promoter. Recombinant riceXIP (rePriceXIP) was secreted into the supernatant and purified to homogeneity with the use of Ni-affinity resin. The molecular mass of rePriceXIP was approximately 44. Read More

Protein Expr Purif 2019 Apr 28;156:44-49. Epub 2018 Dec 28.

State Key Laboratory of Medicinal Chemical Biology, Nankai University, Tianjin, 300353, PR China; Department of Biochemistry and Molecular Biology, College of Life Sciences, Nankai University, Tianjin, 300353, PR China. Electronic address:

The uridine diphosphate glycosyltransferase (UGT) plays the central role in glycosylation of small molecules by transferring sugars to various acceptors including bioactive natural products in plants. UGT89C1 from Arabidopsis thaliana is a novel UGT, a rhamnosyltransferase, specifically recognizes UDP-l-rhamnose as donor. To provide an insight into the sugar specificity for UDP-l-rhamnose and interactions between UGT89C1 and its substrates, the UGT89C1 was expressed in Escherichia coli and purified toward biochemical and structural studies. Read More

Protein Expr Purif 2019 Apr 28;156:25-35. Epub 2018 Dec 28.

School of Pharmacy, Jiangsu University, Zhenjiang, Jiangsu, 212013, China. Electronic address:

Mulberry leaves (Morus alba L.) are of high medicinal value in traditional Chinese medicine with chlorogenic acid (CGA) as its major biologically active constituent. Mulberry leaves require that they be harvested after frost; previous studies have shown CGA accumulation significantly increased after frost. Read More

Protein Expr Purif 2019 Apr 21;156:8-16. Epub 2018 Dec 21.

College of Food Science, Hainan University, Haikou, PR China.

This work attempts to study and optimize the conditions for separating and purifying papain in aqueous two-phase systems (ATPSs). Quaternary ammonium ionic liquids (ILs, 4 wt%) were added as adjuvants to a PEG-phosphate ATPS. On the basis of single-factor experiments, a Box-Behnken design with response surface methodology (BBD-RSM) was used to optimize the purification conditions of papain in the ATPS by setting the NaHPO·2HO concentration, PEG concentration and pH as independent variables and the overall desirability (OD) of the recovery rate of papain, the protein recovery rate and the purification factor as dependent variables. Read More

Protein Expr Purif 2019 Apr 15;156:1-7. Epub 2018 Dec 15.

Department of Biochemistry and Biotechnology, Council of Scientific and Industrial Research (CSIR), Central Leather Research Institute (CLRI), Chennai, India. Electronic address:

Protein engineering is an emerging field for developing novel therapeutic proteins and commercial enzymes, along with a major impact on the global market. In recent decades, advanced methods employing protein modification through expansion of the genetic code have led to the development of proteins with new biochemical and physical properties. These techniques have produced engineered proteins with improved attribute comprising substrate relaxation, protein drug conjugation and high stability under extreme conditions of high temperatures, pH and organic solvents. Read More

Protein Expr Purif 2019 Apr 14;156:36-43. Epub 2018 Dec 14.

School of Biology and Biological Engineering, South China University of Technology, Guangzhou, Guangdong, 510006, China; Guangdong Provincial Key Laboratory of Fermentation and Enzyme Engineering, Guangzhou, 510006, China. Electronic address:

Nuclease P1 gene (nuc P1) which was cloned from Penicillium citrinum and expressed in A. niger Bdel4 with the low-background extracellular protein. The expression strategy of multi-copy nuc P1 in the A. Read More

Protein Expr Purif 2019 Mar 6;155:95-103. Epub 2018 Dec 6.

Laboratory of Plant Research, College of Life Science, Northeast Agricultural University, Harbin, Heilongjiang Province, 150030, PR China. Electronic address:

Dryopteris fragrans (L.) Schott is a traditional herbal medicine containing medicinal sterols and triterpenoids. Squalene synthase (SQS) is the first crucial enzyme in the biosynthesis pathway of sterols and triterpenoids. Read More

Protein Expr Purif 2019 Mar 4;155:120-129. Epub 2018 Dec 4.

University of Applied Sciences Mittelhessen, Institute of Bioprocess Engineering and Pharmaceutical Technology, Wiesenstrasse 14, 35390, Giessen, Germany; Fraunhofer Institute for Molecular Biology and Applied Ecology (IME), Project Group Bioresources, Heinrich-Buff-Ring 26, 35392, Giessen, Germany; Justus Liebig University, Heinrich-Buff-Ring, 35392, Giessen, Germany; Kansas State University, Faculty of Chemical Engineering, 1005 Durland Hall 1701A Platt Street, Manhattan, KS, 66506, USA. Electronic address:

The Cry4AaCter tag is a pull-down tag which promotes the formation of inclusion bodies (IBs) that can be resolubilized in an alkaline buffer. Here, we used the Cry4AaCter tag to create a platform for the production of antimicrobial peptides (AMPs) in Escherichia coli featuring a uniform resolubilization process independent of the peptide fused to the pull-down tag. The Cry4AaCter tag conserves the bioactivity of fusion proteins and thus allows the purification of simple AMPs and more complex AMPs stabilized by disulfide bonds. Read More

Protein Expr Purif 2019 Mar 4;155:104-111. Epub 2018 Dec 4.

State Key Lab of Bioreactor Engineering, New World Institute of Biotechnology, East China University of Science and Technology, Shanghai, 200237, China.

βgalactosidases have wide industrial applications in lactose hydrolysis and transglycosylation reactions. Therefore, there is a need to mine novel and high-quality β-galactosidases with good tolerance and novel features from harsh environments and genomic databases. In this study, an Escherichia coli β-galactosidase-deficient host, ΔlacZ(DE3)pRARE, was constructed by the CRISPR-Cas9 system for screening active β-galactosidases. Read More

Protein Expr Purif 2019 Mar 30;155:130-135. Epub 2018 Nov 30.

College of Life Science and Biotechnology, Shihezi University, Shihezi, 832000, China. Electronic address:

To optimize the expression conditions for human lactoferrin production, we have constructed the transgenic chlorella with human lactoferrin named as GTD8A1-HLF, the original chlorella was separated from Gurbantunggut Desert in Xinjiang China. To further improve the production of human lactoferrin, a sequential methodology was used to optimize human lactoferrin production by GTD8A1-HLF. First, a screening trial using a Plackett-Burman design was done, and variables with statistically significant effects on human lactoferrin bio-production were identified. Read More

Protein Expr Purif 2019 Mar 1;155:86-94. Epub 2018 Dec 1.

Department of Biochemistry and Molecular Biology, MS 200, University of Nevada, Reno, NV, 89557, USA.

This paper presents a simultaneous isolation of pure, intact chloroplasts and mitochondria from mature leaves of Ice plant (Mesembryanthemum crystallinum) and mitochondrial protein preparation for two-dimensional electrophoresis (2DE) analysis under well watered and water -deficit stressed treatments. The washed chloroplasts and mitochondria were purified with Percoll gradients prepared using a Master flex R pump. The chloroplast and mitochondrial proteins were extracted in lysis buffer containing a protease inhibitor mix supplemented with 1 μM Leupeptin and 1 μM E64, followed by precipitation with ice-cold acetone. Read More

Protein Expr Purif 2019 Mar 1;155:112-119. Epub 2018 Dec 1.

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

Bispecific antibodies (bsAbs) are antibodies that can simultaneously bind two distinct targets or epitopes. Their dual-targeting capacity offers expanded therapeutic potential. Currently there is a strong interest in design and production of bsAbs to achieve improved efficacy through novel mechanisms of action. Read More

Protein Expr Purif 2019 Mar 27;155:78-85. Epub 2018 Nov 27.

Centre for Biotechnology and Bioengineering (CeBiB), Department of Chemical Engineering, Biotechnology and Materials, University of Chile, Beauchef 851, Santiago, Chile. Electronic address:

α-Amylase is an endo-acting enzyme which catalyzes random hydrolysis of starch. These enzymes are used in various biotechnological processes including the textile, paper, food, biofuels, detergents and pharmaceutical industries. The use of active enzymes at low temperatures has a high potential because these enzymes would avoid the demand for heating during the process thereby reducing costs. Read More

Protein Expr Purif 2019 Mar 25;155:66-71. Epub 2018 Nov 25.

Faculty of Pure and Applied Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8573, Japan. Electronic address:

Cell surface protein B (CspB) from Corynebacterium glutamicum is used as a pH-responsive peptide tag to enable a simple solid-liquid separation method for isolating a CspB fusion protein. Here we demonstrate the first application of a CspB tag for the purification of Teriparatide, which is a biologic drug that is prescribed for osteoporosis. The Teriparatide was constructed as CspB50TEV-Teriparatide, comprising 50 amino acid residues of CspB, the cleavage site of TEV protease, and Teriparatide. Read More

Protein Expr Purif 2019 Mar 22;155:59-65. Epub 2018 Nov 22.

Department of Genetics & Metabolism, Pasteur Institute of Iran, Tehran, Iran. Electronic address:

Recombinant antibodies have emerged over the last few decades as the fastest growing class of therapeutic proteins for autoimmune diseases. Post-translation modifications of antibodies produced by human cell lines are highly consistent with those existing in natural human proteins and this is a major advantage of utilizing these cell lines. Cinorra is a biosimilar form of the antibody Adalimumab, which is an antagonist of TNF-α used for the treatment of autoimmune diseases. Read More

Protein Expr Purif 2019 Mar 20;155:54-58. Epub 2018 Nov 20.

Division of Biological Sciences, Wonkwang University, Iksan, 54538, South Korea. Electronic address:

The B subunit of Escherichia coli heat-labile toxin (LTB) is a model antigen that induces a strong immune response upon oral administration and enhances immune responses to conjugated and co-administered antigens. We previously examined high expression levels of LTB in plants by chloroplast and synthetic LTB gene expression and found substantially higher expression levels of LTB, compared to nuclear LTB expression in wild-type plants. The 2. Read More

Protein Expr Purif 2019 Mar 19;155:48-53. Epub 2018 Nov 19.

Department of Genomics and Genetic Engineering, Razi Vaccine and Serum Research Institute (RVSRI), Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. Electronic address:

The objective of this study was to introduce a simple and cheap method for purification of flagellin. So, flagellin proteins of Salmonella typhimurium (S. typhimurium), Escherichia coli (E. Read More

Protein Expr Purif 2019 Mar 13;155:27-34. Epub 2018 Nov 13.

Downstream Bioprocessing Laboratory, Department of Life Sciences & Chemistry, Jacobs University, Campus Ring 1, D-28759, Bremen, Germany. Electronic address:

In antibody purification processes, affinity chromatography has been used with Staphylococcus aureus protein A (SpA) as the main ligand. In this work, we present a novel Staphylococcal Protein A (AviPure thereafter), a synthetic ligand analogue based on native SpA B domain, with a molecular weight of approximately 14 kDa. The binding affinity of mAbs to AviPure was evaluated using Surface Plasmon Resonance (SPR) and affinity chromatography methods. Read More

Protein Expr Purif 2019 Mar 26;155:8-14. Epub 2018 Oct 26.

College of Life and Environmental Science, Wenzhou University, Wenzhou, 325035, Zhejiang, China. Electronic address:

Fibroblast growth factor 18 (FGF18) is a member of the fibroblast growth factor family and important in cartilage growth and development. However, the mechanism by which FGF18 mediates its biological functions is still unclear. In our study, we expressed the rhFGF18 protein fused to a HaloTag, (Halo-rhFGF18). Read More

Protein Expr Purif 2019 Mar 8;155:21-26. Epub 2018 Nov 8.

School of Pharmacy, Second Military Medical University, Shanghai, 200433, China. Electronic address:

The interaction between TNF-α with TNFR1 triggers important signaling pathways inducing diverse cellular phenomena including inflammation, apoptosis, etc., and is involved in the pathogenesis and progression of numerous autoimmune diseases. The extracellular domain (ECD) of TNFR has been successfully used to clinically treat such TNF-associated diseases. Read More

Protein Expr Purif 2019 Mar 8;155:43-47. Epub 2018 Nov 8.

School of Pharmaceutical Sciences, Binzhou Medical University, No. 346 Guanhai Road, Yantai, 264003, China. Electronic address:

Nanobodies offer multiple advantages over conventional antibodies in terms of size, stability, solubility, immunogenicity, and production costs, with improved tumor uptake and blood clearance. Additionally, the recombinant expression of nanobodies is robust in various expression systems, such as Escherichia coli, Saccharomyces cerevisiae and Pichia pastoris. P. Read More

Protein Expr Purif 2019 Mar 31;155:1-7. Epub 2018 Oct 31.

Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou, 310014, Zhejiang, China. Electronic address:

Pichia pastoris expression is a mature and efficient eukaryotic expression system. In this work, Aspergillus oryzae lipase (AOL, with the molecular mass of 28 kDa), which can perform highly stereoselective hydrolysis of (R, S)-methyl 2-(4-hydroxyphenoxy) propanoate, was expressed in P. pastoris X-33. Read More

Protein Expr Purif 2019 Feb 31;154:147-151. Epub 2018 Oct 31.

Medical Cellular and Molecular Research Center, Golestan University of Medical Sciences, Gorgan, Iran.

One of a major drawbacks correlated with expressing antibody fragments in bacterial cells is insolubility, which is often regarded as an obstacle in obtaining active molecules. Recombinant proteins aggregated as inclusion bodies within bacterial cells are thought to be unfolded or misfolded, and therefore inactive. So, denaturing and refolding strategies, which are laborious and sometime inefficient, are used to obtain correctly-folded active proteins. Read More

Protein Expr Purif 2019 Feb 28;154:134-139. Epub 2018 Oct 28.

College of Biological Engineering, Henan University of Technology, Zhengzhou, 450001, China. Electronic address:

β-1,3-glucanases, the plant PR-2 family of pathogenesis-related (PR) proteins, can be constitutively expressed and induced in wheat crop to enhance its anti-fungal pathogen defense. This study aimed to investigate the inhibitory effect of wheat β-1,3-glucanase on fungi most commonly associated with wheat kernel. A β-1,3-glucanase from wheat was successfully expressed in Pichia pastoris X-33 and its biochemical and antifungal properties were characterized herein. Read More

Protein Expr Purif 2019 Feb 24;154:140-146. Epub 2018 Oct 24.

Department of Chemistry, Middle Tennessee State University, Murfreesboro, TN, 37132, USA. Electronic address:

A non-specific nucleoside hydrolase has been isolated from germinated Alaska pea seeds. The enzyme catalyzes the hydrolysis of both purines and pyrimidines along with ribo- and deoxyribonucleosides. A purification scheme utilized ammonium sulfate precipitation, ion exchange chromatography and size exclusion chromatography, resulted in 103-fold purification with a recovery of 2. Read More

Protein Expr Purif 2019 Mar 21;155:35-42. Epub 2018 Oct 21.

School of Biological Sciences, Universiti Sains Malaysia, 11800 Minden, Penang, Malaysia; Centre for Chemical Biology, Penang, Malaysia; Malaysian Institute of Pharmaceuticals and Nutraceuticals, NIBM, Malaysia. Electronic address:

Depolymerase is an enzyme that plays an important role in the hydrolysis of polyhydroxyalkanoates [PHAs]. In the current study, Burkholderia cepacia DP1 was obtained from Penang, Malaysia in which the enzyme was purified using ion exchange and gel filtration (Superdex-75) column chromatography. The molecular mass of the enzyme was estimated to be 53. Read More

Protein Expr Purif 2019 Feb 16;154:126-133. Epub 2018 Oct 16.

Department of Food Engineering, Akdeniz University, 07058, Antalya, Turkey; Food Safety and Agricultural Research Center, Akdeniz University, 07058, Antalya, Turkey. Electronic address:

Milk-clotting enzymes used in the dairy industry can be obtained from different sources such as plants, animals, and microorganisms. Recombinant chymosin is the best alternative for the dairy industry due to the differences in physicochemical properties of coagulating enzymes and scarcity of chymosin from animal sources. In this study, glycosylated and non-glycosylated forms of yak chymosin were extracellularly produced in a methylotrophic yeast, Komagataella phaffii (Pichia pastoris). Read More

Protein Expr Purif 2019 Feb 13;154:104-111. Epub 2018 Oct 13.

Department of Biological Sciences, BITS Pilani, K. K. Birla, Goa Campus, NH17B, Zuarinagar, Goa, 403726, India. Electronic address:

The genome of aureophage Phi11 reveals the presence of the gene gp07 which codes for the putative antirepressor protein (GenBank accession no. NC_004615.1). Read More