Development of Extruded Functional Snack Foods for Seniors and Hypocholesterolemic Effect of Hydrolysate from Lactic Fermented Chlorella Produced by High Hydrostatic Pressure-Assisted Protease Hydrolysis(I)

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Project title

Code/計畫編號

MOST109-2221-E229-001-MY2

Translated Name/計畫中文名

銀髮族之擠壓功能性休閒食品的開發與高靜水壓輔助蛋白酶水解乳酸發酵小球藻之水解產物的降膽固醇效果(I)

Funding Organization/主管機關

National Science and Technology Council

Co-Investigator(s)/共同執行人

林雨欣

Department/Unit

Department of Food Science and Technology

Website

https://www.grb.gov.tw/search/planDetail?id=13528982

Year

2020

Start date/計畫起

01-08-2020

Expected Completion/計畫迄

31-07-2021

Co-Investigator(s)

Guan-Wen Chen

Bugetid/研究經費

1225千元

ResearchField/研究領域

食品科技(工)

Description

Abstract

電漿之蝕刻技術於工業上用於去除表面材料，將此與超高壓加工技術結合並用於綠藻細胞壁上，可以釋放更多活性物質。蛋白質經酵素水解後，其水解物中之胜肽具有抑制 HMG-CoA reductase (HMGR) 的活性，使甲羥戊酸的合成受阻，進而降低膽固醇合成。另一方面，胜肽能降低在微胞中飲食來源的膽固醇的溶解度，破壞膽酸對於膽固醇的乳化作用，進而抑制膽固醇微胞形成，以達到降膽固醇的吸收效果。因此，本研究以高壓與電漿進行破壁處理後的綠藻為原料，將破壁之綠藻以乳酸菌發酵後再以高壓輔助商業酵素水解，產生富含功能性生物活性肽的蛋白質水解物，最後將此水解物製作成銀髮族機能性膨發休閒食品。研究計畫擬以三年期間進行，第一年依先前科技部的研究成果，篩選出最佳破壁結果之小球藻及加入乳酸菌發酵的最佳發酵時間後，再篩選各種商業酵素對於小球藻蛋白質水解率、抑制 HMGR 活性以篩選出最佳的水解物組別，並分析其抑制膽固醇微胞形成的能力。最後，再將此水解物進行腸胃道的安定性試驗。第二年將利用腸胃道消化試驗後之蛋白質水解物進一步以膠體層析並分析其分子量的分佈與各劃分物之 HMGR 抑制能力。再利用 RP-HPLC 純化 HMGR 抑制活性最高之胜肽，並經由蛋白質 N 端定序分析其胜肽序列。另外，以飲食誘導形成之高膽固醇血症大鼠對水解物之降膽固醇效果進行評估。第三年將依動物實驗結果評估成人之建議攝取量，開發成米果，並評估其儲藏安定性試驗。藉此研究期望能以小球藻為原料，開發成具有降膽固醇效果又適合銀髮族食用之銀髮族機能性膨發休閒食品。 Etching of plasma technology is used in in industry to remove surface material of package. Hence, we’d like to combine plasma and high pressure processing (HPP) to break cell wall of Chlorella, to release more bioactive compounds. Peptides from protein hydrolysates have the inhibitory activity against HMG-CoA reductase to reduce the formation of cholesterols by obstructing the synthesis of mevalonic acid/mevalonate pathway. On the other hand, peptides can reduce the solubility of cholesterols from food in micelles, disrupting the emulsification of cholic acid toward cholesterols, to inhibit the formation of micelles and decrease cholesterols level. In this study, using Chlorella after HPP and plasma as material, cell wall broken of Chlorella will be fermented by lactic acid bacteria and then hydrolyzed by HPP-assisted protease, to produce protein hydrolysate containing functional bioactive peptide as a reference for the development of functional snack foods for seniors. This study will be completed in three years. In the first year, we aim to find the best hydrolysis condition and lactic fermentation time of commercial protease by investigating the protein hydrolysis rate and the HMG-CoA reductase inhibitory activity base on the previous research results of Ministry of Science and Technology. In additional, this hydrolysate was analyzed for its ability to the inhibition percentages of cholesterol micellar solubility, and tested for in vitro gastrointestinal digestion . During the second year, according to stability of the active peptides of in vitro gastrointestinal digestion assay, molecular weight distribution and the HMGR inhibition capacity of each fraction will be analyzed. The peptide with the highest inhibitory activity of HMGR will be purified by reverse-phase HPLC and identified its sequence; Moreover, the Chlorella protein hydrolysate is used in animal experiment, to investigate the inhibition effect in high cholesterol diet induced rats. Finally, the recommended intake of dose was evaluated based on the results of animal experiments will develop into functional snack foods for seniors and storage stability test of the product will be evaluated. We expect that Chlorella could be the raw material, and develop extruded functional snack foods for seniors.

Keyword(s)

綠藻
電漿
超高壓加工技術
細胞壁破壁
蛋白質水解物
活性胜肽
羥甲基戊二酸單醯輔酶 A 還原酶
微胞
Chlorella；
Plasma
High pressure processing
Cell wall-breaking
Protein hydrolysate
Bioactive peptide
HMG-CoA reductase
Micelles