Targeted degradation of gluten proteins in wheat flour by prolyl endoprotease and its utilization in low immunogenic pasta for gluten sensitivity population

Gluten is responsible for adverse immune effects and causes wheat allergy, celiac disease, and other wheat-related disorders to genetically predisposed individuals. Wheat flour protein immunogenicity can be reduced by proteolytic degradation enzymes. The present study aims to utilize Aspergillus niger prolyl endoprotease (EC 3.4.21.26) to specifically cleave the proline-rich sequences and degradation of gluten proteins in the wheat flour. The Triticum aestivum (HD-2851, NIAW-917), Triticum durum (UAS-428), Triticum dicoccum (DDK- 1025) were taken for proteolytic degradation using AN-PEP. The enzymatic modification was carried out in an optimum condition (pH-4.0, at 40 °C) for 5 h. The wheat flour gliadin content was reduced up to 90–95% in HD-2851, DDK-1025, NIAW-917 and UAS-428 shown up to 74% reduction of gliadins. The complete hydrolysis of gliadins (α, β, ω, ϒ) and glutenin subunits were confirmed by using immunoblot, ELISA, RP-HPLC chromatograph, anti-PEP-I (PSQQQQQPK), anti-PEP-II (LGQQQPFPPQQPYPQPQPFK) and anti-HMW-GS antibodies. The modified wheat flour was taken for the processing of pasta with the replacement of 30% chickpea flour. The gluten content of MWF pasta was found to be 48 ppm which is 99.95% lesser than the control pasta. Sensory evaluation revealed that the pasta was acceptable with overall quality when compared with control pasta.