Enhancement of Tryptic Digestibility of Milk β-Lactoglobulin Through Treatment with Recombinant Rice Glutathione/Thioredoxin and NADPH Thioredoxin Reductase/Thioredoxin Systems

Abstract

β-Lactoglobulin (BLG), a member of lipocalin family, is one of the major bovine milk allergens. This protein exists as a dimer of two identical subunits and contains two intramolecular disulfide bonds that are responsible for its resistance to trypsin digestion and allergenicity. This study aimed to evaluate the effect of reduction of disulfide bonds of BLG with different rice thioredoxins (Trxs) on its digestibility and allergenicity. Therefore, the active recombinant forms of three rice Trx isoforms (OsTrx1, OsTrx20, and OsTrx23) and one rice NADPH-dependent Trx reductase isoform (OsNTRB) were expressed in Escherichia coli. Based on SDS-PAGE, HPLC analysis, and competitive ELISA, the reduction of disulfide bonds of BLG with OsNTRB/OsTrx23, OsNTRB/OsTrx1, GSH/OsTrx1, or GSH/OsTrx20 increased its trypsin digestibility and reduced its immunoreactivity. The finding of this study opens new insights for application of plant Trxs in the improvement of food protein digestibility. Especially, the use of OsTrx20 and OsTrx1 are more cost-effective than E. coli and animal Trxs due to their reduction by GSH and no need to NADPH and Trx reductase as mediator enzyme.

Animal‐derived food allergen: A review on the available crystal structure and new insights into structural epitope

“…The reduction of disulfide bonds destabilizes the native rigid globular structure of β‐lactoglobulin, making it more accessible to enzymatic hydrolysis. Meanwhile, the disruption of the exposed disulfide bond (Cys 66 –Cys 160 ) changes the conformation and accessibility of the major human IgE epitope (Shahriari‐Farfani et al., 2019). The disulfide bonds in bovine serum albumin not only fortify the tertiary structure but also stabilize its major antigenic site that is exposed for binding (Jaiswal & Worku, 2021).…”

Thioredoxin-1: A Promising Target for the Treatment of Allergic Diseases

“…Accordingly, Trx1-rich yeast extract can potentially be used to ferment foods, such as alcoholic beverages and bread. Recently, recombinant Trx1 rice has been shown to improve β-lactoglobulin digestion and decrease its allergenicity, thereby improving the feasibility and practicality of its large-scale application; a plant Trx system would be more cost-effective than those of Escherichia coli or animals ( 45 ).…”