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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53778

Title: 乳酸菌生物防腐劑組合輔劑應用於烘焙食品防腐之研究
Study on the Application of Lactic Acid Bacterial Biopreservatives with Adjuncts on Bakery Foods Preservation
Authors: Lu, Szu-Hsaun
呂思璇
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 防腐;細菌素;苯乳酸;劑輔;烘焙食品;乳酸菌
Preservation;bacteriocin;phenyllactic acid;adjunct;baked foods;lactic acid bacteria
Date: 2019
Issue Date: 2020-07-03T08:43:46Z
Abstract: 本篇研究主旨為研發新型乳酸菌生物防腐劑,與輔劑 EDTA-Na2 及乳酸鈉 (Sodium lactate) 組合,以提升於烘焙食品防腐之應用效果。Lactococcus lactis BCRC10791 於起始 pH 6.0 在 30oC 環境中培養 30 hr,乳酸鏈球菌素 (Nisin) 含量為 5.11 mg/mL 即 5,110 IU/mL。產片球菌素 A (Pediocin A) 之乳酸菌 Pediococcus pentosaceus BCRC 14024 胞外物質上清液 (Cell-free suspension, CFS),經高效液相層析 (High performance liquid chromatography, HPLC) 分子篩型管柱收集後,因經高壓反應後呈現斷裂之結果;而經快速蛋白層析儀 (Fast Performance Liquid Chromatography, FPLC) 收集後以 HPLC 分子篩型管柱判定為 88.49 kDa 和87.60 kDa。KP3、BCRC10791 和 BCRC14024 單獨及組合對 Escherichia coli BCRC10675 和 Bacillus cereus BCRC10446 致死濃度 (Minimum bactericidal concentration, MBC) 為 25-50 mg/mL,對 Aspergillus neoniger BCRC32126 和 Penicillium roqueforti BCRC30874 之 MBC 則為 50-200 mg/mL。將產苯乳酸 (Phenyallatic acid, Ph-LA) 之乳酸菌分離菌株 KP3 (30oC、60 hr、起始 pH 6.0)、KP4 (37oC、42 hr、起始 pH 6.0)、DP2 (30oC、60 hr、起始 pH 5.0)、K8 (30oC、30 hr、起始 pH 5.0) 和產 Nisin 之乳酸菌 BCRC 10791 (30oC、30 hr、起始 pH 6.0) 及產 Pediocin 之乳酸菌 BCRC14024 (30oC、60 hr、起始 pH 5.0) 之 CFS 組合輔劑乙二胺四乙酸二鈉 (Disodium ethylenediaminetetraacetate, EDTA-Na2) 和乳酸鈉,調整起始 pH 值至 pH 5.0、6.0 和 7.0,以液態抗菌法測試對菌株 E. coli 和 B. cereus 生長之相對抑制活性 (Relatively inhibition activity, RIA),結果顯示 CFS 組合 20 mM EDTA-Na2相較於未添加 EDTA-Na2 有顯著提高 RIA 之效果,而0.5% 乳酸鈉之添加能部分提高 RIA。將 5 倍濃縮之 CFS (Ph-LA、Nisin和 Pediocin 1:1:1 之CFS 凍乾粉末回溶至原體積 1/5) 組合輔劑後,杯子蛋糕烘烤前噴灑 (1) 75 µL EDTA-Na2、(2) CFS 25 µL組合 50 µL EDTA-Na2、(3) CFS 和 EDTA-Na2 各 75 µL、(4) CFS、EDTA-Na2 和乳酸鈉各 25 µL、(5) CFS、EDTA-Na2 和乳酸鈉各 75 µL 有均較佳防腐結果,皆能在 25oC 下保存達成大於 15 天之保存期限 (較控制組延長 9 天以上)。而小餐包於烘烤前或烘烤中噴灑 (1) CFS、EDTA-Na2 和乳酸鈉各 25 µL 和 (2) CFS、EDTA-Na2 和乳酸鈉各 75 µL 呈現最佳防腐效果,於第 7 天出現黴菌菌絲 (較控制組延長 3 天)。綜合上述之實驗結果,烘烤前及烘烤中噴灑 5 倍濃縮 CFS 組合輔劑噴灑於烘焙產品表面,防腐效果比烘烤後再噴灑為佳,也初步證明 CFS 組合輔劑於高溫處理後呈現較高的防腐活性。
The purpose of this study is to develop a new type of lactic acid bacteria biological preservative, and to explore the combination with the adjuncts EDTA-Na2 and sodium lactate to enhance the preservation of bakery food. Lactococcus lactis BCRC10791 was cultured at 30oC for 30 hr at an initial pH 6.0 and a nisin content was 5.11 mg/mL, 5,110 IU/mL. BCRC 14024 cell-free supernatant (CFS) produce Pediocin A, collected by high performance liquid chromatography (HPLC) molecular sieve column The results of the fracture after high pressure reaction were collected by fast performance liquid Chromatography (FPLC) and determined by HPLC molecular sieve column to be 88.49 kDa and 87.60 kDa. Minimal bactericidal concentration (MBC) of CFS derived from strains KP3, BCRC10791 and BCRC14024 alone and in combination against Escherichia coli BCRC10675 and Bacillus cereus BCRC10446 are 25-50 mg/mL, and their minial fungicidal concentration (MFC) against Aspergillus neoniger BCRC32126 and Penicillium roqueforti BCRC30874 are 50-200 mg/mL. Phenyallatic acid (Ph-LA) lactic acid bacteria isolate KP3 (30oC, 60 hr, initial pH 6.0), KP4 (37oC, 42 hr, initial pH 6.0), DP2 (30oC, 60 hr, initial pH 5.0), K8 (30oC, 30 hr, initial pH 5.0), Nisin-producing lactic acid bacteria BCRC 10791 (30oC, 30 hr, initial pH 6.0) and Pediocin-producing lactic acid bacteria BCRC14024 (30oC, 60 hr, initial pH 5.0) cell-free supernatant (CFS) combined adjuncts disodium ethylenediaminetetraacetate (EDTA-Na2) and sodium lactate (SL), adjusted initial pH values to pH 5, 6 and 7. The relative inhibitory activity (RIA) against the growth of strains E. coli and B. cereus was tested by liquid antibacterial method. The results showed that the CFS combination of 20 mM EDTA-Na2 significantly improved the effect of RIA compared to the absence of EDTA-Na2, while the addition of 0.5% SL partially increase RIA. Add 5-fold concentrated CFS (Ph-LA, Nisin and Pediocin 1:1:1 CFS freezed powder back to the original volume 1/5) combination adjuncts, cup cake sprayed (1) 75 µL EDTA-Na2, (2) 25 µLCFS + 50 µL EDTA-Na2, (3) CFS and EDTA-Na2 each 75 µL, (4) CFS + EDTA-Na2 + SL each 25 µL, (5) CFS + EDTA-Na2 + SL each 75 µL before baking, which shelf life of more than 15 days saved at 25oC (more than 9 days longer than the control group). The buns was sprayed with CFS + EDTA-Na2 + SL each 25 µL and CFS + EDTA-Na2 and SL each 75 µL before baking or during baking to give the best preservation. Mold appeared in 7 days (3 days longer than the control group). Based on the above experimental results, 5-fold concentrated CFS combination adjuncts sprayed on the surface of the baked product before baking and during baking are better than spraying after baking. It is also preliminarily proved that CFS combination adjunts exhibits higher preservation after high temperature treatment.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010732022.id
http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53778
Appears in Collections:[食品科學系] 博碩士論文

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