BACKGROUND/OBJECTIVESDue to its beneficial health effects, use of buckwheat has shown a continuous increase, and concerns regarding the allergic property of buckwheat have also increased. This study was conducted for evaluation of the hydrolytic effects of seven commercial proteases on buckwheat allergens and its allergenicity.MATERIALS/METHODSExtracted buckwheat protein was hydrolyzed by seven proteolytic enzymes at individual optimum temperature and pH for four hours. Analysis was then performed using SDS-PAGE, immunoblotting, and competitive inhibition ELISA (ciELISA) with rabbit antiserum to buckwheat protein, and direct ELISA with pooled serum of 21 buckwheat-sensitive patients.RESULTSAlkaline protease, classified as serine peptidase, was most effective in reducing allergenicity of buckwheat protein. It caused decomposition of the whole buckwheat protein, as shown on SDS-PAGE, and results of immunoblotting showed that the rabbit antiserum to buckwheat protein no longer recognized it as an antigen. Allergenicity showed a decrease of more than 50% when pooled serum of patients was used in ELISA. Two proteolytic enzymes from Aspergillus sp. could not hydrolyze buckwheat allergens effectively, and the allergenicity even appeared to increase.CONCLUSIONSSerine-type peptidases appeared to show a relatively effective reduction of buckwheat allergenicity. However, the antigenicity measured using rabbit antiserum did not correspond to the allergenicity measured using sera from human patients. Production of less allergenic buckwheat protein may be possible using enzymatic hydrolysis.
In order to develop a new starter for fermented milk, Lactobacillus paracasei subsp. paracasei BFI46 (BFI46) obtained from new-born infant feces was investigated for physiological characteristics. Good immunomodulating activity was evident compared with commercial lactic acid bacteria starter cultures. The optimum growth temperature of BFI46 was 40 o C with 12 h required to reach pH 4.3. Testing with 13 different antibiotics revealed greatest sensitivity of BFI46 to penicillin-G and chloramphenicol, and heightened resistance to neomycin, kanamycin and polymyxin. BFI46 displayed higher esterase activities compared to 18 other enzymes, was comparatively tolerant to bile juice and able to survive at pH 2 for 3 h, and displayed high resistance against Escherichia coli and Salmonella Typhimurium with a survival rate of 57.14% and 96.36%, respectively. The results indicate that BFI46 could be an excellent starter culture for fermented milk with high level of immunomodulating activity.
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