- 學位論文
乳酸菌發酵酒粕產物對美白與抗氧化作用之探討
Studies on whitening and antioxidation effect analysis from lees fermented by lactic acid bacteria
摘要
本論文針對三種酒粕(米酒、清酒及啤酒)以實驗室篩選之益生性乳酸菌及以Lactobacillus rhamnosus GG (LGG)當作對照組發酵24小時，發酵的產物進行美白及抗氧化之分析。在美白測試上，所有發酵產物大都能抑制市售mushroom tyrosinase的活性。但在降低人類黑色素瘤細胞(A375 cell line)內黑色素含量上，卻只有兩個啤酒酒粕發酵產物(BE33及BE38)可以降低黑色素的生成，其分別降低18%及17%的黑色素含量。當發酵液的凍乾粉末溶液濃度為5 mg/ml時，能與陽性對照組熊果素(40 mM)具有類似抑制黑色素細胞內tyrosinase活性的能力(約28％的抑制能力、熊果素32％)和降低黑色素的生成(25~27%、熊果素29％)。在抗氧化效果上，酒粕發酵產物(米酒、清酒及啤酒)分別進行清除DPPH自由基、還原力、亞鐵離子螯合能力、ABTS自由基清除能力及細胞內ROS的反應等試驗。在清除DPPH能力上，各發酵產物均具有清除DPPH的能力。啤酒發酵液的清除能力相當於對照組BHA (1 mg/ml)清除能力。米酒及清酒發酵液的清除能力相當於BHA清除能力約2倍。在還原能力上，各發酵產物均具有還原效果。以SE7(清酒E7)、RV12(米酒V12)及RN1(米酒N1)發酵液的清除活性較強，相當於對照組BHA (1 mg/ml)清除能力的約2倍。但各酒粕發酵液均無鐵離子螯合能力效果。在ABTS自由基清除能力上，各發酵產物均具有清除ABTS的能力。啤酒發酵液的清除能力相當於對照組trolox (1 mM)清除能力的5倍。米酒及清酒發酵液的清除能力相當於trolox清除能力約7倍。在細胞內去除ROS上，有2個清酒酒粕發酵液(SE7及SV12)與陽性對照組維生素C (1 mM)有類似的消除細胞內活性氧能力。整體而言，米酒及清酒酒粕發酵液比啤酒酒粕發酵液的抗氧化效果高。
並列摘要
In this study, six strains isolated from local Taiwanese fermented vegetables, which could tolerate low pH conditions and high concentration of bile salts were selected for fermented three kind of lees (rice lees, sake lees and beer lees). The inhibition effects of tyrosinase activity, melanin accumulation ability and antioxidant activity of the LAB-fermented products were determined. In whitening effect, all fermented products could inhibit the activity of mushroom tyrosinase. But, only two fermented products (BE33 and BE38) could decrease melanin accumulation with 18% and 17% in A375 human malignant melanoma cells. These fermented products were frozen and adjusted as 5 mg/ml, then cultured the A375 cells at the same condition. We find the intracellular activity of tyrosinase was decrease with 28% and melanin accumulation was decrease with 25~27%. The effect of whitening was the same as the positive control, arbutin (40 mM). In antioxidation effect, the antioxidation activities of all fermented products were determined by measuring their α-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging ability, reducing ability, ferrous ion chelating ability, ABTS radical scaveng- ing ability, and intracellular ROS scavenging ability. In DPPH radical scavenging ability, all fermented products could scavenge DPPH radical. The effect of beer lees fermented products were the same as BHA (1 mg/ml), and rice and sake lees fermented products were 2-folds as BHA. In reducing ability, all fermented products had reducing ability. Three fermented products (SE7, RV12 and RN1) had stronger reducing ability, and their reducing ability were 2-folds as BHA. In ABTS scavenging ability, all fermented products could scavenge ABTS radical. The effect of beer lees fermented products were 5-folds as trolox (1 mM), and rice and sake lees fermented products were 7-folds as trolox. In intracellular ROS scavenging ability, two fermented products (SE7 and SV12) had intracellular ROS scavenging ability and the effects were similar to the control, vitamin C (1 mM). But, no chelating effect on ferrous ion was found in all fermented products. Overall, rice and sake lees fermented products had better antioxidant effect than that of beer lees fermented products.