乳酸菌固態發酵黑豆產品之機能性

Bioactivities of Black Soybean by Lactic Acid Bacteria Solid-State Fermentation

李佩穎(Pei-Yin Lee)

指導教授 ： 蔣丙煌

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2005年)

https://doi.org/10.6342/NTU.2005.02350

摘要

中文摘要
黑豆學名為【Glycine max (L.) Merr】，為黃豆的黑皮種，具有許多的營養及機能性成分，在中國古代醫書的記載中佔有很重要的地位，可說是一個藥用食品。過去的研究中指出，食品基質經過乳酸菌發酵可以有效的提升其機能性，例如抗氧化、抗癌、抗發炎等。本實驗的第一部分為，利用黑豆為發酵基質，觀察乳酸菌固態發酵的生長情況包括pH、菌數、還原醣及peptide濃度在發酵期間的變化。以乳酸菌進行固態發酵之後，進行四種抗氧化活性之測試(清除DPPH自由基、清除超氧陰離子及清除過氧化氫)並以抗氧化活性的結果來做為指標，篩選不同的菌種(BCRC 12250 Lactobacillus plantarum, BCRC 10695 Lactobacillus acidophilus, BCRC 14602 Bifidobacterium longum, BCRC 11846 Bifidobacterium breve)、發酵時間(3, 5, 7 days) 及發酵基質(不經酵素水解, 0.4% 纖維素酶 (V), 0.4% 蛋白酶 (P), 0.2% 纖維素酶(V) + 0.2% 蛋白酶(P))。研究結果顯示，在纖維素酶及蛋白酶水解的基質中以L. acidophilus為發酵菌種發酵三天的樣品之抗氧化能力最好。在第二部分的實驗中則延續第一部份的結果，再進一步對此發酵產品進行抗癌及抗發炎的測試；抗癌的實驗中，以八株不同細胞株進行篩選，發現AGS胃癌細胞株對樣品最為敏感，之後的實驗則以其為標靶，對不同區分物進行檢測利用。MTT及細胞周期的檢測發現不同的區分物中以Hex層對誘導細胞的凋亡最為有效。而在抗發炎的實驗中利用RAW 264.7的實驗模式探討細胞前發炎激素(TNF-α)的改變及NO生成量的變化，可發現樣品的水粗萃物及水區分層對未被激活的巨噬細胞有促進其產生TNF-α的能力，而對LPS/IFN-γ激活的細胞中 之NO產生之抑制以Hex及BtOH區分層效果最好。

關鍵字

黑豆 ； 乳酸菌 ； 固態發酵

並列摘要

Black soybean is a kind of soybean with dark seed coat. In ancient books and records of Chinese herbs, black soybean frequently played an important role. Black soybean contains many biofunctional compounds and possesses antitumer activity. Since past researches has indicated that the bioactivity of substrates could enhanced after lactic acid bacteria fermentation, the purposes of this project were to investigate the feasibility of using solid-state fermentation of lactic acid bacteria using black soybean as substrate, and to evaluate the bioactivities of the fermented products.
The growth condition of lactic acid bacteria were envaluated by observing the changes of pH and cell during fermentation. The result displayed that lactic acid bacteria grew well in the enzymatic hydrolyzed black soybean, but not in the un-treated substrates. In the antioxidant activity, three assay methods were used for determining the antioxidant activities of the fermented black soybean product by different lactic acid bacteria (Lactobacillus plantarum BCRC 12250, Lactobacillus acidophilus BCRC 10695, Bifidobacterium longum BCRC 14602, Bifidobacterium breve BCRC 11846) for various time periods (3, 5, 7 days), and in different types of media (non-enzymatic hydrolyzed medium (N), 0.4% Viscozyme hydrolyzed medium (V), 0.4% Flavozyme: Alcalase=3:2 hydrolyzed medium (P), 0.2% Viscozyme + 0.2% Flavozyme+Alcalase=3:2 hydrolyzed medium (V+P)). Our result indicated that L. acidophilus 3- day-fermentation showed the highest antioxidant ability and was chosen for subsequent studies on anticancer and anti-inflammatory activity.
In the anticancer tests, eight kinds of cancer line have been tested, and the gastric cancer cell line, AGS, had the highest sensitivity to the fermented samples. Our results also showed that the fraction extracted by Hexane of L. acidophilus 3-day-fermented product was most effective in inhibiting the proliferation of AGS cell line. In the anti-inflammatory test, crude water extract and WA phase could increase production of TNF-λrom unstimulated RAW 264.7, and the Hex fraction and BtOH fraction had the strongest ability to inhibite of the production of NO from LPS/IFN-γactivated RAW 264.7.

並列關鍵字

solid-state fermentation ； black soybean ； lactic acid bacteria

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