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Economical Production and antioxidant function of exopolysaccharides from Lactobacillus acidophilus La and Bifidobacterium adolescentis Ba
|關鍵字:||胞外多醣體;Exopolysaccharides;抗氧化活性;生產可能曲線;中草藥;乳化指數。;antioxidant activity;production possibility curve;Chinese medicinal herbs;emulsification index.||出版社:||食品暨應用生物科技學系所||引用:||Abd el-Gawad IA, el-Sayed EM, Hafez SA, el-Zeini HM, and Saleh FA, 2004, Inhibitory effect of yoghurt and soya yoghurt containing bifidobacteria on the proliferation of Ehrlich ascites tumour cells in vitro and in vivo in a mouse tumour model, British Journal of Nutrition, 92:81–86. Audy SJ, Roy DL, and LaPointe G, 2010, Sugar source modulates exopolysaccharide biosynthesis in Bifidobacterium longum subsp longum CRC 002, Microbiology, 156: 653–664. Batdorj B, Trinetta V, Dalgalarrondo M, Prevost H, Dousset X, Ivanova I, Haertle T, and Chobert JM, 2007, Isolation, taxonomic identification and hydrogen peroxide production by Lactobacillus delbrueckii subsp. lactis T31, isolated from Mongolian yoghurt: inhibitory activity on food-borne pathogens, Journal of Applied Microbiology, 103:584–593. 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具抗氧化活性的胞外多醣（EPS）是乳酸菌（LAB）重要的經濟產物，因為它可以賦予對人體健康的有益作用。本研究主要利用嗜酸乳桿菌LA(LaP）和雙歧桿菌Ba（BaP）發酵生產EPS企圖提高其經濟生產率，並加強其體外的抗氧化作用，以促進保健效果。實驗以MRS培養基培養菌株La和Ba發酵萃取EPS作為對照組，而以MRS含葡萄糖、乳糖、果糖及化學組成已知的優良E培養基等分別作為實驗組，以比較對照組和實驗組EPS之產量和生產率。EPS的糖類成分以高效液相色層分析法（HPLC）配備RI檢測器進行量化測定。抗氧化活性測定包括清除1,1-二苯基-2-肼（DPPH）自由基的能力，抑制亞麻油酸過氧化能力、亞鐵離子螯合能力，以及鐵還原力測定等。另外EPS添加每升10.0克粉末中草藥（CMH）的DPPH自由基清除能力也被評估其促進保健效果的潛力，而EPS添加十種中草藥的乳化指數也分別被測定。為經濟生產EPS目的而發展新穎的生產可能曲線模式(PPC model)，以獲得最佳培養基為90% NB與50% MRS broth的使用組合，實驗分析結果發現EPS的最高生產率與濃度在LaP分別為15.5±0.8%, 388.4±6.2毫克/升而在BaP為14.3±0.7%, 342.3±2.3毫克/升，是以化學組成已知的E培養基所產生的。糖質組成分析顯示LaP與BaP皆以葡萄糖為主，最高濃度分別為5.2±0.4毫克/毫升與4.6±0.7毫克/毫升，比較參考方法分析的結果分別為4.75毫克/毫升與4.69毫克/毫升則相當。抗氧化潛力分析，LaP與BaP分別顯示90.3±0.1% 和90.2±0.5% DPPH自由基清除活性。在37oC培養24 h後，LaP與BaP抑制亞麻油酸過氧化分別降低到32.0±7.9% and和44.5±1.5%。還原能力分別為0.29± 0.04%和0.28±0.06%。亞鐵離子螯合作用則分別為75.8±5.0%和80.0±6.7%。此外，所有中草藥(10.0 g/L)添加EPS(10 g/L)的溶液對DPPH自由基清除能力均大於50％，顯示EPS可增強CMH的抗氧化活性，具有作為食品補給劑的應用潛力。而乳化指數（％）分析結果CHM添加EPS大於未添加EPS的CHM且超過50％，顯示胞外多醣體LaP與BaP可提高中草藥的乳化效果，可作為商業應用的食品添加劑。故利用控制培養基組成的策略不僅可以提高EPS的生產率和產量，也可產生不同功能結構的EPS，因此二種不同的胞外多醣LaP與BaP皆具有抗氧化潛力，在休閒食品保健產業上具有作為食品補給劑的良好潛勢。
Exopolysaccharides (EPS) of lactic acid bacteria (LAB) with antioxidant activities are economically important as it can confer beneficial effects on human health. The main objective of this study is to improve the productivity of EPS from Lactobacillus acidophilus La (LaP) and Bifidobacterium adolescentis Ba (BaP), and to characterize the antioxidant effects of EPS in vitro, for enhancing health-promoting effects. The Man-Rogosa-Sharpe (MRS) broth was used to incubate stains La and Ba to harvest EPS as a control group. The MRS broth with various nutrition compositions like glucose, lactose, fructose and E medium were used as experimental groups, respectively. The productivity and yield of EPS were estimated for comparison between control and experimental groups. The carbohydrate composition of LaP and BaP were quantified by high performance liquid chromatography (HPLC) equipped with RI detector. Emulsification index of Chinese medicinal herbs (CMH) with EPS additives were separately determined. A novel model of production possibility curve was approached to the optimal productivity set of NB (90%) and MRS (50%) broth for the purpose of economical production. The highest yield and EPS concentration of LaP and BaP were respectively 15.5�0.8%, 388.4�6.2 mg/L, and 14.3�0.7%, 342.3�2.3 mg/L, produced by strains La and Ba that were incubated in MRS+E medium. Analysis of sugar composition showed that the main sugar was glucose with concentrations of 5.2 � 0.4 mg/ml in LaP (10 mg/mL) and 4.6�0.7 mg/ml in BaP (10 mg/mL), which were similar to the results of reference method that were respectively 4.75 mg/ml and 4.69 mg/ml. Regarding antioxidant potential of EPS, a concentration of 10 mg/mL of LaP and BaP solution, respectively exhibited 90.3�0.1% and 90.2�0.5% DPPH radical-scavenging activity. After 24 h incubation with LaP and BaP at 37 oC, linoleic acid peroxidation was reduced to 32.0�7.9% and 44.5�1.5%, respectively. The reducing capacities of LaP and BaP were 0.29�0.04% and 0.28�0.06%, respectively. The ferrous ion chelating effects of LaP and BaP were 75.8�5.0% and 80.0�6.7%, respectively. In addition, all the effects of CMH solution (10.0 g/L) with 10 mg/mL of EPS additives on DPPH radical scavenging capabilities were larger than 50%, which with antioxidative property could merit its potential application in food supplement. All the emulsification index of CMH with 4.0% EPS additives were larger than that of CMH and over 50%, which revealed that both LaP and BaP can improve the emulsion effects of CMH available to be as food additives for commercial application. Current strategy of controlling the composition of medium not only improved the productivity and yield of EPS, but also transformed EPS to acquire different structure with specific functionality. Hence, both LaP and BaP with antioxidant potential may be useful as supplements in the health-promoting food of leisure industry.
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