Preprint Article Version 1 This version is not peer-reviewed

Functional Expression and Characterization of the Recombinant N-acetyl-glucosamine/N-acetyl-galactosamine-Specific Marine Algal Lectin BPL3

Version 1 : Received: 28 November 2017 / Approved: 28 November 2017 / Online: 28 November 2017 (07:20:39 CET)

A peer-reviewed article of this Preprint also exists.

Hwang, H.-J.; Han, J.-W.; Kim, G.H.; Han, J.W. Functional Expression and Characterization of the Recombinant N-Acetyl-Glucosamine/N-Acetyl-Galactosamine-Specific Marine Algal Lectin BPL3. Mar. Drugs 2018, 16, 13. Hwang, H.-J.; Han, J.-W.; Kim, G.H.; Han, J.W. Functional Expression and Characterization of the Recombinant N-Acetyl-Glucosamine/N-Acetyl-Galactosamine-Specific Marine Algal Lectin BPL3. Mar. Drugs 2018, 16, 13.

Journal reference: Mar. Drugs 2018, 16, 13
DOI: 10.3390/md16010013

Abstract

Lectins, characterized by their carbohydrate-binding ability, have an extensive practical application. However, their industrial use is limited by low yields, and few active recombinant lectins have been reported. In this study, the algal lectin BPL-3 (Bryopsis plumosa lectin 3) was successfully produced using a bacterial expression system, BL21(DE3), with an artificial repeated structure (dimeric construct). Recombinant dimeric BPL3 (rD2BPL3) was confirmed by LC-MS/MS spectrometry. Expression efficiency was greater for the construct with the repeat structure (rD2BPL3) than the monomeric form (rD1BPL3). Optimal conditions for expression were 1 mM IPTG at 20 °C. Recombinant lectin was purified under denaturing conditions and refolded by the flash dilution method. Recombinant BPL3 was solubilized in 1× PBS containing 2 M urea. rD2BPL3 showed strong hemagglutination activity using human erythrocytes, similar to that of native BPL3. rD2BPL3 had a similar sugar specificity to that of the native protein, i.e., to N-acetyl-glucosamine (GlcNAc) and N-acetyl-galactosamine (GalNAc). Glycan array results showed that recombinant BPL3 and native BPL3 exhibited different binding properties. Both showed weak binding activity to α-Man-Sp. Native BPL3 showed strong binding specificity to the alpha conformation of amino sugars, and rD2BPL3 had binding activity to the beta conformation. The process developed in this study was suitable for the quality-controlled production of high amounts of soluble recombinant lectins.

Subject Areas

Bryopsis plumosa; BPL3; lectin; hemagglutinin; recombinant; tandem repeat; GlcNAc; GalNAc

Readers' Comments and Ratings (0)

Leave a public comment
Send a private comment to the author(s)
Rate this article
Views 0
Downloads 0
Comments 0
Metrics 0
Leave a public comment

×
Alerts
Notify me about updates to this article or when a peer-reviewed version is published.