[發明專利]富含多磷酸化多肽的酪蛋白水解物和制備方法

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• 申請號: 201010269516.8
• 申請日: 2010-09-02
• 公開（公告）號: CN101948524A
• 公開（公告）日: 2011-01-19
• 發明（設計）人: 蘇榮欣;齊崴;何志敏
• 申請（專利權）人: 天津大學
• 主分類號: C07K14/47
• 分類號: C07K14/47;C07K1/36;C07K1/34;C07K1/30;C07K1/18;C12P21/06
• 代理公司: 天津市杰盈專利代理有限公司 12207
• 代理人: 王小靜

1.一種富含多磷酸化多肽的酪蛋白水解物，其特征在于它是以酪蛋白為原料，采用蛋白水解酶進行適度水解，水解度需控制10％以內，沸水浴加熱滅酶，采用5kDa超濾膜從反應體系中分離出各種產物多肽，再將該產物用1kDa的超濾膜進行截留，所得截留液采用鈣-乙醇選擇性沉淀磷酸肽，再將其用去離子水或揮發性鹽溶液溶解，上樣到強陽離子交換柱上，分離獲得帶多個磷酸化的多肽，采用噴霧干燥或冷凍干燥獲得產品。

2.根據權利要求1所述的酪蛋白水解物，其特征在于所述的多肽均含有多個磷酸化基團，共檢出9個多磷酸化多肽，占水解產物中所有磷酸肽的30％，每個多肽所帶磷酸化基團至少4個。

3.根據權利要求1所述的酪蛋白水解物，其特征在于所述的多肽的組成與氨基酸序列：

α_S160-98(5P)Met-Glu-Ala-Glu-SerP-Ile-SerP-SerP-SerP-Glu-Glu-Ile-Val-Pro-Asn-SerP-Val-Glu-Gln-Lys-His-I

?????????????le-Gln-Lys-Glu-Asp-Val-Pro-Ser-Glu-Arg-Tyr-Leu-Gly-Tyr-Leu-Glu-Gln-Leu

α_S21-19(4P)?Lys-Asn-Thr-Met-Glu-His-Val-SerP-SerP-SerP-Glu-Glu-Ser-Ile-Ile-SerP-Gln-Glu-Thr

α_S21-20(4P)?Lys-Asn-Thr-Met-Glu-His-Val-SerP-SerP-SerP-Glu-Glu-Ser-Ile-Ile-SerP-Gln-Glu-Thr-Tyr

α_S21-24(4P)?Lys-Asn-Thr-Met-Glu-His-Val-SerP-SerP-SerP-Glu-Glu-Ser-Ile-Ile-SerP-Gln-Glu-Thr-Tyr-Lys-G

?????????????ln-Glu-Lys

β1-24(4P)???Arg-Glu-Leu-Glu-Glu-Leu-Asn-Val-Pro-Gly-Glu-Ile-Val-Glu-SerP-Leu-SerP-SerP-SerP-Glu-Glu-

?????????????Ser-Ile-Thr

β1-25(4P)???Arg-Glu-Leu-Glu-Glu-Leu-Asn-Val-Pro-Gly-Glu-Ile-Val-Glu-SerP-Leu-SerP-SerP-SerP-Glu-Glu-

?????????????Ser-Ile-Thr-Arg

β1-28(4P)???Arg-Glu-Leu-Glu-Glu-Leu-Asn-Val-Pro-Gly-Glu-Ile-Val-Glu-SerP-Leu-SerP-SerP-SerP-Glu-Glu-

?????????????Ser-Ile-Thr-Arg-Ile-Asn-Lys

β1-29(4P)???Arg-Glu-Leu-Glu-Glu-Leu-Asn-Val-Pro-Gly-Glu-Ile-Val-Glu-SerP-Leu-SerP-SerP-SerP-Glu-Glu-

?????????????Ser-Ile-Thr-Arg-Ile-Asn-Lys-Lys

β1-32(4P)???Arg-Glu-Leu-Glu-Glu-Leu-Asn-Val-Pro-Gly-Glu-Ile-Val-Glu-SerP-Leu-SerP-SerP-SerP-Glu-Glu-

?????????????Ser-Ile-Thr-Arg-Ile-Asn-Lys-Lys-Ile-Glu-Lys

????????????????????????????????????????????????????????????????????????????????????????????????????。