Direct immobilization and recovery of recombinant proteins from cell lysates by using EctP1-peptide as a short fusion tag for silica and titania supports

Kim, Jong Ki; Abdel‐Hamid, Mohamed; Pack, Seung Pil

doi:10.1016/j.ijbiomac.2019.05.105

Cited by 27 publications

(24 citation statements)

References 62 publications

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“…The absorbance at 562 nm was measured using the BCA method to determine the protein content (Kim, Abdelhamid, & Pack, 2019). Amino acid content was determined using the ninhydrin colorimetric method, and absorbance at 570 nm was measured (Zhang, Fu, Zhou, Kang, & Li, 2013).…”

Section: Methodsmentioning

confidence: 99%

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Effect of different drying temperatures on the rehydration of the fruiting bodies of Yu Muer (Auricularia cornea) and screening of browning inhibitors

Chen

Liu

et al. 2020

Food Science & Nutrition

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cornea cv. Yu Muer) is a new white variety of edible fungus that was selected from a mutant of Auricularia cornea by the Engineering Research Center of the Ministry of Education, Jilin Agricultural University. Yu Muer is in genus Auricularia, family Auriculariaceae, order Auriculariales, class Agaricomycetes, and phylum Basidiomycota. It is an edible fungus and is also used in medicine (Royse, 2014; Wang, Jiang, et al., 2019a; Wang, Li, et al., 2019b). The fruiting bodies of Yu Muer are thick, tender, and crispy tastes like jellyfish, and have a jade-like warm, soft color. It is rich in nutrients, including physiologically active substances such as polysaccharides,

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Section: Methodsmentioning

confidence: 99%

“…The absorbance at 562 nm was measured using the BCA method to determine the protein content (Kim, Abdelhamid, & Pack, 2019).…”

Section: Determination Of Ppo Activity and Nutrient Composition In mentioning

confidence: 99%

Effect of different drying temperatures on the rehydration of the fruiting bodies of Yu Muer (Auricularia cornea) and screening of browning inhibitors

Chen

Liu

et al. 2020

Food Science & Nutrition

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“…Using these amino acids has the additional advantage of stabilizing eluted proteins [39, 40]. Eluting with salts such as NaCl and MgCl 2 (1 M) was not possible and seems to be bothersome even at high concentrations (1–5 M) [15, 17, 41]. Lysine's and arginine's competitive elution effects work at moderate concentrations of around 0.5 M but were also reported to elute at lower concentration (0.1 M) [20].…”

Section: Resultsmentioning

confidence: 99%

“…Purification of a protein with a silica affinity tag was previously optimized for GFP regarding silica type, buffers, and pH [20, 29, 41]. Therefore, Davisil 643 silica particles were chosen and Tris buffer at pH 8.0.…”

Section: Resultsmentioning

confidence: 99%

Purification of a peptide tagged protein via an affinity chromatographic process with underivatized silica

Rauwolf

Steegmüller

Schwaminger

et al. 2021

Engineering in Life Sciences

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Silica is widely used for chromatography resins due to its high mechanical strength, column efficiency, easy manufacturing (i.e. controlled size and porosity), and low‐cost. Despite these positive attributes to silica, it is currently used as a backbone for chromatographic resins in biotechnological downstream processing. The aim of this study is to show how the octapeptide (RH)4 can be used as peptide tag for high‐purity protein purification on bare silica. The tag possesses a high affinity to deprotonated silanol groups because the tag's arginine groups interact with the surface via an ion pairing mechanism. A chromatographic workflow to purify GFP fused with (RH)4 could be implemented. Purities were determined by SDS‐PAGE and RP‐HPLC. The equilibrium binding capacity of the fusion protein GFP‐(RH)4 on silica is 450 mg/g and the dynamic binding capacity around 3 mg/mL. One‐step purification from clarified lysate achieved a purity of 93% and a recovery of 94%. Overloading the column enhances the purity to >95%. Static experiments with different buffers showed variability of the method making the system independent from buffer choice. Our designed peptide tag allows bare silica to be utilized in preparative chromatography for downstream bioprocessing; thus, providing a cost saving factor regarding expensive surface functionalization. Underivatized silica in combination with our (RH)4 peptide tag allows the purification of proteins, in all scales, without relying on complex resins.

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“…Solid-binding peptides/proteins have been of great interest in many applications such as surface functionalization, fusion tags for protein purification, fabrication of functional architectures, drug delivery, and biological immobilization of enzymes (Ikeda et al, 2009(Ikeda et al, , 2010Coyle and Baneyx, 2014;Kim et al, 2019;Hellner et al, 2020). Silica is one of the most abundant materials in nature, widely used in structure materials, food industry, pharmaceutical applications, and semiconductors.…”

Section: Introductionmentioning

confidence: 99%

Ultrahigh Adhesion Force Between Silica-Binding Peptide SB7 and Glass Substrate Studied by Single-Molecule Force Spectroscopy and Molecular Dynamic Simulation

Zhang

Chen

et al. 2020

Front. Chem.

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Many proteins and peptides have been identified to effectively and specifically bind on certain surfaces such as silica, polystyrene and titanium dioxide. It is of great interest, in many areas such as enzyme immobilization, surface functionalization and nanotechnology, to understand how these proteins/peptides bind to solid surfaces. Here we use single-molecule force spectroscopy (SMFS) based on atomic force microscopy to directly measure the adhesion force between a silica-binding peptide SB7 and glass surface at single molecule level. SMFS results show that the adhesion force of a single SB7 detaching from the glass surface distributes in two populations at ~220 pN and 610 pN, which is higher than the unfolding forces of most mechanically stable proteins and the unbinding forces of most stable protein-protein interactions. Molecular dynamics simulation reveals that the electrostatic interactions between positively charged arginine residues and the silica surface dominates the binding of SB7 on silica. Our study provides experimental evidence and molecular mechanism at the single-molecule level for the SB7-based immobilization of proteins on silica-based surface, which is able to withstand high mechanical forces, making it an ideal fusion tag for silica surface immobilization or peptide-base adhesive materials.

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Direct immobilization and recovery of recombinant proteins from cell lysates by using EctP1-peptide as a short fusion tag for silica and titania supports

Cited by 27 publications

References 62 publications

Effect of different drying temperatures on the rehydration of the fruiting bodies of Yu Muer (Auricularia cornea) and screening of browning inhibitors

Effect of different drying temperatures on the rehydration of the fruiting bodies of Yu Muer (Auricularia cornea) and screening of browning inhibitors

Purification of a peptide tagged protein via an affinity chromatographic process with underivatized silica

Ultrahigh Adhesion Force Between Silica-Binding Peptide SB7 and Glass Substrate Studied by Single-Molecule Force Spectroscopy and Molecular Dynamic Simulation

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