Ito Masako, Kato Takeo, Matsuda Tsukasa
Aichi Industrial Technology Institute, Food Research Center, Nagoya, Japan.
Biosci Biotechnol Biochem. 2005 Jun;69(6):1137-44. doi: 10.1271/bbb.69.1137.
The rice grains (RG) and rice seed proteins remaining in rice miso were investigated with a view point to the potential allergenicity of rice miso. RG ranging from 36 to 180 mg dry weight per g dry miso were separated from several samples of commercially available rice miso. Scanning electron microscopy of the recovered RG indicated that starch granules disappeared almost completely while protein bodies remained intact in RG. Most of the major seed proteins were extracted from RG by heating with 1% SDS/2% 2-mercaptoethanol and detected by SDS-polyacrylamide gel electrophoresis. Major rice allergenic proteins, 14-16 kDa albumin (Alb14-16) and alpha-globulin (alpha-Glb) were also detected by immunoblotting using the specific antisera, and their contents were estimated to be 1.7 to 9.0 and 1 to 7 mg protein per g dry RG respectively. However, the major rice proteins, including glutelin and prolamin, in RG were insoluble in salt, alcohol, and urea solutions, but soluble in 6 M guanidine hydrochloride (Gu-HCl). By immunoblotting and ELISA, no Alb14-16 and only a slight amount of alpha-Glb were detected even in the 6 M Gu-HCl fraction, indicating that these major allergenic proteins are denatured and are present in an insoluble form in rice miso.
从大米味噌中残留的米粒（RG）和大米种子蛋白的角度出发，对大米味噌的潜在致敏性进行了研究。从市售大米味噌的几个样品中分离出每克干味噌中干重为36至180毫克的RG。对回收的RG进行扫描电子显微镜观察表明，淀粉颗粒几乎完全消失，而蛋白质体在RG中保持完整。通过用1%十二烷基硫酸钠/2% 2-巯基乙醇加热从RG中提取了大部分主要种子蛋白，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳进行检测。还使用特异性抗血清通过免疫印迹检测到主要的大米致敏蛋白，14-16 kDa白蛋白（Alb14-16）和α-球蛋白（α-Glb），其含量估计分别为每克干RG 1.7至9.0毫克蛋白和1至7毫克蛋白。然而，RG中的主要大米蛋白，包括谷蛋白和醇溶蛋白，不溶于盐、醇和尿素溶液，但可溶于6 M盐酸胍（Gu-HCl）。通过免疫印迹和酶联免疫吸附测定，即使在6 M Gu-HCl组分中也未检测到Alb14-16，仅检测到少量的α-Glb，这表明这些主要致敏蛋白已变性并以不溶性形式存在于大米味噌中。
