促脂解大豆胜肽之最適水解條件、序列及其應用

專利類型

發明

專利國別 (專利申請國家)

中華民國

專利申請案號

100121652

專利證號

I 407915

專利獲證名稱

促脂解大豆胜肽之最適水解條件、序列及其應用

專利所屬機關 (申請機關)

東海大學

獲證日期

2013/09/11

技術說明

本技術可應用酵素膜反應系統生產具有促脂解作用的大豆肽及提供後續純化促脂解肽的方法。依此系統所篩選的酵素，其最適水解條件可將分離大豆蛋白 (ISP) 水解並提升其促脂解作用，即此ISP水解物 (ISPH) 可使3T3-L1脂肪細胞的丙三醇釋放量最大化，丙三醇釋放量愈高表示ISPH的促脂解作用愈高；依照最適水解條件製備促脂解活性的ISPH ，利用膜限值1,000-100,000 daltons 的濾膜進行區分，所篩選的濾膜分子量限值，亦可進一步的使ISPH增加3T3-L1 脂肪細胞的丙三醇的釋放量。進一步將篩選的濾膜區分物透過膠體層析純化，獲得分子量介於 189-2,080 Da胜肽片段 (GF3) 可進一步顯著提升其脂解活性 (p<0.05) 。最後利用兩段式逆相層析分離 GF3 獲得單一胜肽片段 RHF4-2 與 RHF4-3具有顯著的促脂解活性 (p<0.05) ，與控制組比較可分別提升 84% 與 95 % 。 The technique can produce soy peptides with lipolysis-stimulating (LS) action from enzymatic hydrolysis of isolated soy protein (ISP) using membrane reactor system. The technique also provides several methods to purify the LS peptides in ISP hydrolysate (ISPH). The selected enzyme with its optimum hydrolysis conditions can hydrolyze ISP to produce the hydrolysate with the maximum glycerol release in 3T3-L1 adipocytes. Higher glycerol release represents higher LS action. Moreover, several membranes have been used to fractionate the hydrolysate into several membrane fractions. The selected membrane fraction can further enhance the amount of glycerol release in the cells and will be purfied by gel chromatography and two steps of reverse phase chromatography to obtaine RHF4-2 and RHF4-3 which show significantly increase by 84% and 95 % LS activity, respectively, as compared with the control (p<0.05).

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04-23594711

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