雙叉桿菌胞內物對活性氧之抗致突變性與抗氧化性

Antimutagenic and antioxidative activities of intracellular extracts of bifidobacteria against reactive oxygen species

許雅鈞(Ya-Jiun Sheu)

指導教授 ： 游若篍

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2004年)

https://doi.org/10.6342/NTU.2004.01920

摘要

本研究探討六株雙叉桿菌 Bifidobacterium adolescentis BCRC14606、B. bifidum BCRC14615、B. breve BCRC11846、B. infantis BCRC14602、B. lactis Bb-12 與 B. longum BCRC14634 對活性氧物質的抗致突變性。使用安氏試驗法之測試菌株 Salmonella typhimurium TA102 來分析雙叉桿菌胞內物對氧化突變劑 tert-butylhydroperoxide (t-BOOH) 與過氧化氫 (H2O2) 誘導突變之防禦效果。結果顯示六株雙叉桿菌菌體懸浮液 (1010 CFU/mL) 經超音波破碎，離心 (15,000 × g, 30 min) 移除細胞碎片後所得到的胞內物，對 t-BOOH 和 H2O2 之抗致突變性分別為 15.9-50.9 % 和 51.5-72.3 %。更進一步研究結果顯示抗致突變性的主要機制為去致突變作用。而雙叉桿菌的抗氧化變異性可能歸因於其抗氧化能力，所以進一步評估其抗氧化性，檢測系統包括還原力、螫合亞鐵離子能力、DPPH 自由基及活性氧物質之清除能力探討。結果顯示六株雙叉桿菌之胞內物都表現出良好的還原力，以 B. lactis 表現最好，其還原力相當於 328 μg/mL 的抗壞血酸。清除 DPPH 自由基測定中，則以 B. longum 清除能力最好。螫合亞鐵離子方面，6 株雙叉桿菌胞內物表現出 34-52 % 之螫合力，以B. longum最佳。在清除活性氧物質方面，胞內物均具有清除超氧陰離子之能力，清除率介於 52.7-69.9 %，而以 B. adolescentis 效果最好；對過氧化氫之清除率則在 5.0-18.0 % 之間，其中以 B. bifidum有較佳表現；而在清除羥基自由基方面，B. bifidum 具有 11.8 % 之清除力，為 6 株雙叉桿菌中表現最佳者。

關鍵字

雙叉桿菌 ； 抗致突變性 ； 抗氧化性 ； 活性氧物質

並列摘要

Antimutagenic activities of intracellular extracts of Bifidobacterium adolescentis BCRC14606, B. bifidum BCRC14615, B. breve BCRC11846, B. infantis BCRC14602, B. lactis Bb-12 and B. longum BCRC14634 against reactive oxygen substances were studied by a modified Ames test using Salmonella typhimurium TA102 as a test strain. The intracellular extracts were prepared by disrupting cells (1010 CFU/mL) with an ultrasonic disrupter and subsequently removing cell debris by centrifugation at 15,000 × g for 30 min. Assays for the ability of intracellular extracts of bifidobacteria against mutations induced by peroxide mutagens tert-butylhydroperoxide (t-BOOH, 10μg/plate) and hydrogen peroxide (H2O2, 6μmol/plate) toward S. typhimurium TA102 were developed. Results showed that the antimutagenic activities of bifidobacteria against t-BOOH and H2O2 were 15.9-50.9% and 51.5-72.3%, respectively. It was further shown that main mechanism of antimutagenicity is desmutagenic effect. The antimutagenic activities of bifidobacteria were probably attributed to their antioxidative activities. Therefore, antioxidative activities of these bifidobacteria were further determined by measuring their reducing activity, ferrous ion chelating activity,

並列關鍵字

antioxidative activity ； bifidobacteria ； antimutagenicity ； reactive oxygen species

參考文獻

陳惠英、顏國欽。1996。膳食中之抗致突變物及其作用機制。中華民國營養學會雜誌。21:323-338。

Aherne SA, O’Brien NM. 2000. Mechanism of protection by the flavonoids, quercetin and rutin, against tert-butylhydroperoxide and menadione-induced DNA single strand breaks in Caco-2 cells. Free Radic Biol Med 29:507–514.

Allameh A. 1997. Comparison of the effect of low- and high-dose dietary butylated hydroxytoluene on microsome-mediated aflatoxin B1- DNA binding. Cancer Lett 114:217-220.

Ames BN, McCann J, Yamasaki E. 1975. Methods for detecting carcinogens and mutagens with the Salmonella/mammalian- microsome mutagenicity test. Mutat Res 31:347-364.

Ames BN. 1989. Endogenous DNA damage as related to cancer and aging. Mutat Res 214:41-46.