Pea Seedling Amine Oxidase Application: an Emerging Antihistamine Strategy in Tuna Fish

Abstract

Despite the known antihistaminic effects of PSAO, this enzyme remains unrecognized as an antihistamine additive in the food industry. The objective of this study was to ascertain the plausible histamine-degrading effects of a new food-grade additive (pea seedling extract) in tuna fish. PSAO was purified from cultured pea seedlings via ionexchange and size-exclusion chromatography. SDS-PAGE results indicated that the PSAO subunit has an apparent molecular weight of about 95 kDa. To evaluate the optimized conditions for antihistaminic activity of the enzyme, the reaction of PSAO and histamine was carried out in KPi buffer under conditions relevant to the fish processing industry. 1 unit mL -1 of PSAO significantly reduced the histamine content at pH 7.0 but not at pH 5.0. Accordingly, its antihistaminic activity was assessed in homogenized skipjack tuna fish (Katsuwonus pelamis) using an ion-paired HPLC method. The results showed that PSAO can reduce 87.14% of the histamine content of the tuna fish at pH 7.0, 37°C. This study indicates that PSAO -purified or even filtrate of pea seedling homogenate -can degrade histamine, which suggests practical application of this enzyme in fish processing industry.Pea seeds (Pisum sativum L) were purchased from Nunhems Zaden BV, Haelen, Netherlands. The seeds, disinfected with a solution of sodium hypochlorite, were cultured in plastic plug-trays with a soilless media of cocopeat and perlite (1:1 v/v). The etiolated (dark-grown) seedlings were harvested above the roots after 12-days germination at room temperature. After harvesting, immediate purification of PSAO was carried out at 4°C.

Carboxymethyl starch/alginate microspheres containing diamine oxidase for intestinal targeting

“…In order to further identify the protein profiles, samples of free DAO and of DAO extracted from microspheres were run in SDS‐PAGE under nonreducing conditions for protein staining and for zymography. Previous studies indicated the presence of a single band at 72 kDa in denaturating SDS‐PAGE for vegetal DAO extracted from L. sativus and a band at 95 kDa for DAO extracted from Pisum sativum L . The molecular mass of pea seedling amine oxidase was also investigated using the method of gel filtration, indicating a value of 184.0 ± 2.6 kDa .…”

“…a single band at 72 kDa in denaturating SDS-PAGE [40,43] for vegetal DAO extracted from L. sativus and a band at 95 kDa for DAO extracted from Pisum sativum L [44]. The molecular mass of pea seedling amine oxidase was also investigated using the method of gel filtration, indicating a value of 184.0 ± 2.6 kDa [45].…”

In vitro determination of diamine oxidase activity in food matrices by an enzymatic assay coupled to UHPLC-FL

“…Among the range of methodologies described in the literature that challenge the determination of DAO activity, the majority are based on the measurement of the liberation of hydrogen peroxide or the consumption of oxygen occurring along the oxidative deamination reaction [13,21,[24][25][26][27][28][29][30]. Those largely used approaches face an important drawback, as the presence of hydrogen peroxide and dioxygen may be markedly influenced by the concomitant presence of other enzymatic capacities in certain complex biological matrices [34,35].…”

“…A wide range of methods to detect in vitro DAO activity are described in the literature. With the aim of measuring the rate of substrate degradation or the generation of by-products of this enzymatic reaction, most methods are based on the detection of hydrogen peroxide, aldehyde or dioxygen by spectrophotometric [13,21,24,25], fluorometric [26], polarographic [27,28] or amperometric [29,30] techniques. Radioimmunoassay techniques have also been extensively described, consisting of the radioactive labeling of the substrate and the scintillation counting of its consumption [3,11,31].…”