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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53712

Title: 龍膽石斑魚卵蛋白質鑑定及其潛在活性胜肽分析
Analysis of Potential Bioactive Peptides from Giant Grouper (Epinephelus lanceolatus) Roe Protein
Authors: Fenny Crista Anastasia Panjaitan
柯芬妮
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 血管緊張素轉換酶(ACE-I);抗氧化劑;龍膽石斑魚卵;電腦模擬;體外實驗;蛋白質體學
Angiotensin-I converting enzyme (ACE-I);antioxidant;giant grouper roe;in silico;in vitro;proteomics
Date: 2019
Issue Date: 2020-07-03T08:43:03Z
Abstract: 乾燥的龍膽石斑魚卵中主要含有的蛋白質如卵黃蛋白原(from Epinephelus coioides; NCBI accession number: AAW29031.1)、apolipoprotein A-1的前驅物(from Epinephelus coioides; NCBI accession number: ACI01807.1)和apolipoprotein E (from Epinephelus bruneus; NCBI accession number: AEB31283.1),可透過蛋白質體學技術分析其特徵(SDS-PAGE, in-gel digestion, mass spectrometry and on-line Mascot database analysis)。這些蛋白質可利用BLAST和BIOPEP-UWM數據庫在電腦中進行分析,透過BLAST進行序列相似性比對,顯示Epinephelus coioides和Epinephelus lanceolatus的卵黃蛋白原序列有70%的相似性。此外,水解過程中會產生許多胜肽,可透過BIOPEP-UWM數據庫預測其潛在的生物活性,而胃蛋白酶(pH> 2)被預測為最有潛力用於製備活性牲肽的酵素。透過抑制ACE 實驗分析得知,在酵素濃度為2 mg/ml時,以胃蛋白酶(34.09 ± 1.83%)水解龍膽石斑魚卵的ACE- I抑制效果為最佳,比利用鳳梨蛋白酶(31.15 ± 4.65%)和木瓜蛋白酶(21.51 ± 1.41%)水解時抑制性高,然而,木瓜蛋白酶和鳳梨蛋白酶水解的魚卵分別表現出比胃蛋白酶水解產物具更好的DPPH清除能力(21.36 ± 0.37%)和還原性(0.151 ± 0.040%),整體而言,透過電腦預測結果與本研究中的實驗分析結果相互呼應,因此,此研究證實龍膽石斑魚卵具有作為營養保健品或藥品的潛力。
Major proteins contained in dried giant grouper roe such as vitellogenin (from Epinephelus coioides; NCBI accession number: AAW29031.1), apolipoprotein A-1 precursor (from Epinephelus coioides; NCBI accession number: ACI01807.1) and apolipoprotein E (from Epinephelus bruneus; NCBI accession number: AEB31283.1) were characterized through compiled proteomics techniques (SDS-PAGE, in-gel digestion, mass spectrometry and on-line Mascot database analysis). These proteins were subjected to in silico analysis using BLAST (Basic Local Alignment Search Tool) and BIOPEP-UWM database. Sequence similarity search by BLAST revealed that the aligned vitellogenin sequences from Epinephelus coioides and Epinephelus lanceolatus share 70% identity. Moreover, prediction of potential bioactivities through BIOPEP-UWM database resulted in numerous bioactive peptides. Pepsin (pH >2) was predicted to be the most promising enzyme for the production of bioactive peptides. In vitro analysis confirmed that pepsin hydrolysates of giant grouper roe exhibited the highest ACE-I inhibition (34.09 ± 1.83%) compared to roe hydrolysates obtained from bromelain (31.15 ± 4.65%) and papain (21.51 ± 1.41%) proteolysis at 2 mg/ml concentration. However, roe hydrolyzed by papain and bromelain showed better DPPH scavenging activity (21.36 ± 0.37%) and reducing power activity (0.151 ± 0.040%), respectively, than did pepsin hydrolysates. Overall, the prediction through in silico approach was corresponded to in vitro analysis in this study. It thus confirmed the potentiality of giant grouper roe as raw material for the generation of nutraceutical or pharmaceutical ingredients.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010532078.id
http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53712
Appears in Collections:[食品科學系] 博碩士論文

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