Cloning, Expression, and the Effects of Processing on Sarcoplasmic-Calcium-Binding Protein: An Important Allergen in Mud Crab

• DOI: 10.1021/acs.jafc.7b02381
• Publication Date: 2017-07-10T17:47:27Z
• Authors: Meng-Jun Hu; Guang-Yu Liu; Yang Yang; Tzu-Ming Pan; Yi-Xiang Liu; Le-Chang Sun; Min-Jie Cao; Guang-Ming Liu
• Keywords: Scylla paramamosain; Cross-reactivity

Shellfish allergy is a prevalent, long-lasting disorder usually persisting throughout life. However, the allergen information incomprehensive in crab. This study aimed to identify novel crab, show its potential diagnosis and reduce allergenicity by food processing. A 21-kDa protein was purified from Scylla paramamosain confirmed as sarcoplasmic calcium binding (SCP) matrix-assisted laser desorption ionization-time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF-MS). Total RNA isolated crab muscle, rapid amplification of cDNA performed obtain an ORF 579 bp that coded for 193 amino acid residues. According results circular dichroism analysis ELISA assay, recombinant SCP (rSCP) expressed Escherichia coli showed similar physicochemical immunoreactive properties native (nSCP). Additionally, extensive cross reactivity among different species bidirectional IgE cross-reactivity between nSCP rSCP were detected iELISA. The reduced via Maillard reaction or enzymatic cross-linking reaction, which scanning electron microscopy, dot blot, digestion assay. straightforward reproducible way developed high yields maintains structural integrity full reactivity, could compensate low specific IgE-titers most patient sera future diagnosis. Furthermore, effective approaches production hypoallergenic seafood.