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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/15152

Title: Bacillus subtilis YJ-3生產蛋白酶之條件探討及開發機能性大豆胜肽鈣乳酸菌發酵產品
Optimal Conditions for the Production of Proteases from Bacillus subtilis YJ-3 and Development of Lactic Acid Bacteria Fermented Soybean Products with Calcium
Authors: Hsin-Hsien Hou
侯信獻
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: Bacillus subtilis;大豆蛋白;乳酸菌;胜肽;抗氧化;
Bacillus subtilis;soy protein;Lactic acid bacteria;peptide;Calcium
Date: 2008
Issue Date: 2011-06-30T08:04:22Z
Abstract: 本研究為利用培養基組成份不同,誘導Bacillus subtilis YJ-3生產選擇性胞外酵素水解大豆溶液,併用乳酸菌Lactobacillus plantarum BCRC 10069發酵24小時,探討發酵過程中大豆蛋白水解情形,進一步分析可溶性蛋白質、胜肽及總游離胺基酸含量,並探討發酵產物經不同分子量劃分後與金屬鈣離子螯合情形,藉以研發富含機能性螯合金屬離子胜肽飲品。結果顯示Bacillus subtilis YJ-3 在複合培養基 (含skim milk 1.0%、soya meal 1.0%、sucrose 1.0%、NaCl 0.5%、K2HPO4 0.25%) 於37oC、150 rpm 震盪培養3天後,中、鹼性蛋白酶活性為680.6、567.9U/mL. min。利用該酵素液於50oC 水解大豆蛋白溶液6小時,pH由 7.38降至5.81,加入 0.1% 蔗糖經乳酸菌 (LAB) 發酵24小時後,其可溶性蛋白質由23.9 mg/mL增加至 24.6 mg/mL、胜肽含量由 5.1 mg/ml上升到 12.3 mg/ml及總游離胺基酸含量由 2.6 mg/mL上升至 4.4 mg/mL,顯示乳酸菌在發酵過程中可降解蛋白質。大豆蛋白經胞外液水解6小時及併用Lactobacillus plantarum BCRC 10069 發酵24小時,再利用切向流式超過濾系統將濾液依分子量大小劃分MW為 <1 kDa、1-5 kDa及 >5 kDa,進行噴霧乾燥,則每克可溶性蛋白質其鈣含量分別為 139.03、71.60及 50.91mg,噴乾後胜肽鈣粉分子量小於 1 kDa的抗氧化能力為最強,其清除 DPPH 能力最高為94%,總抗氧化能力 (TEAC) 為 3.50 mM 且還原力最高,顯示胜肽鈣粉具抗氧化機能性。根據上述實驗結果顯示大豆蛋白經 Bacillus subtilis YJ-3 酵素液水解及併用乳酸菌發酵後,能有效使蛋白質水解,以 <1 kDa 以下的胜肽其螯合鈣離子能力最佳且抗氧化能力亦最強。
Abstract The purpose of this study was to investigate the optimal media for Bacillus subtilis YJ-3 to produce proteases and its application in the hydrolysis of soybean proteins, which could consequently enhance their functionalities. The optimal medium for Bacillus subtilis YJ-3 was investigated based on the composition of tryptic soy broth (TSB). The highest neutral (680.6 U/mL•min) and alkaline (567.9 U/mL•min) proteases activities were observed in that incubated in modified medium containing 1% skim milk, 1% soybean meal, 1% sucrose, 0.5% NaCl, 0.25% K2HPO4 after 72 hr incubation at 37oC, 150 rpm. The pH of hydrolysates of soybean decreased from 7.38 to 5.81 after 6 hr hydrolysis using selective enzymes of 3 days incubated Bacillus subtilis YJ-3 at 50oC. During 6 hr hydrolysis, peptides and total free amino acids significantly increased, compared with those without hydrolysis. The soluble proteins and peptides increased from 23.9 to 24.6 mg/mL and 5.1 to 12.3 mg/mL, respectively (P<0.05), while free amino acids increased from 2.6 to 4.4 mg/mL after 24 hr LAB fermentation at 37oC. The peptides with MW less than 1000 Da after 24 hr LAB fermentation could chelate calcium (139.3 mg/g), while those between 1000 and 5000 Da could only chelate 71.60 mg/g. The peptides with MW higher than 5000 Da could only chelate 50.91 mg of calcium/g of soluble proteins or peptides. According to the results obtained from this study, the peptides with MW <1000 Da showed the highest DPPH scavenging ability, calcium chelating activity, TEAC and reducing power.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0M95320033
http://ntour.ntou.edu.tw/ir/handle/987654321/15152
Appears in Collections:[食品科學系] 博碩士論文

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