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. 2023 Nov 22;16(12):100845.
doi: 10.1016/j.waojou.2023.100845. eCollection 2023 Dec.

Fructose biphosphate aldolase: A new cassava allergen

Anne K R M Ventura  1   2   3 , Safiri de P Alves  1   2   3 , Roberta A Castro  3   4 , Bruno C Rossini  5 , Lucilene S Delazari  5   6 , Amanda M de Oliveira  1   2 , Ana I S Moretti  1   2 , Fabio F M Castro  3   4   7 , Jorge Kalil  1   2   3   6 , Ariana C Yang  3   4 , Keity S Santos  1   2   3   7
Affiliations

Fructose biphosphate aldolase: A new cassava allergen

Anne K R M Ventura et al. World Allergy Organ J. .

Abstract

Background: Food allergy has considerably increased in recent years and this situation has been aggravated mainly by the consumption of more processed and complex foods, since minor or potentially allergenic foods are not required to be labeled. Manihot esculenta (cassava) is a widely consumed food in South America, Africa, and Asia and can be used in the production of flour and starch, as well as several other products. This root can cause allergic reactions with symptoms ranging from mild to severe.

Methods: Thus, the aim of this study was the characterization of the immunogenic cassava proteins responsible for sensitizing patients allergic to it. Using a 2D-SDS-PAGE based proteomic approach, six proteins were identified, including Fructose Bisphosphate Aldolase (FBA). Recombinant FBA was produced in Expi293 cells and evaluated by immunoblotting with the serum of 10 individual study subjects.

Results: Our results showed six cassava IgE-reactive proteins. From those, recombinant fructose bisphosphate aldolase (FBA) showed a positivity of 80% among tested sera, proving to be a highly sensitizing protein.

Conclusion: The recombinant FBA molecule obtained in this study can be important for in vivo diagnostic assays, by producing more accurate results, and for desensitization protocols, in which the use of the isolated molecule produces more precise results by avoiding secondary sensitization.

Trial registration: All patients signed a consent form approved by the internal ethics committee CAPPesq, Comissão de Ética para Análise de Projetos de Pesquisa do HC FMUSP (CAAE: 10420619.6.0000.0068).

Keywords: Food hypersensitivity; Fructose-biphosphate aldolase; Manihot; Proteome.

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Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Fig. 1
Fig. 1
SDS-PAGE and Immunoblot analysis of cassava extract. (a) Cassava extract (E) on reducing SDS-PAGE. Immunoblot of individual allergic patients' sera,, , , , , , , , , second antibody control (2nd) and patients non-allergic (C1: non-atopic and C2: atopic) (b) 2D-SDS-PAGE of cassava protein extract (I) and Western blotting (II) showing the IgE binding of pooled sera, , , , , , , , , to cassava protein. pI, isoelectric point; MW, molecular weight, kDa, kiloDalton
Fig. 2
Fig. 2
FBA (fructose-bisphosphate aldolase) recombinant protein (R) on reducing SDS-PAGE. Immunoblot of individual allergic patients' sera (n = 10). MW, molecular weight, kDa, kiloDalton
Fig. 3
Fig. 3
Gastric simulation with pepsin the protein fructose biphosphate aldolase in SDS gel - PAGE 12 % acrylamide. MM: Molecular marker; C: undigested protein control with pepsin; times: 10 min, 30 min and 60 min - incubation time at 37 °C with pepsin enzyme; kDa: kiloDalton
See this image and copyright information in PMC

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