- 學位論文
由芥菜發酵物中分離鑑定益生菌及其特性分析
Isolated, Identification, and Characterization of Potential Probiotic Lactic Acid Bacteria from Sawi Asin
摘要
芥菜是一種印尼人常吃的傳統非乳製品發酵物。和乳製品發酵物相比，芥菜的發酵是不需額外添加菌種的，是個微生物來源豐物的傳統發酵食物。然而，從芥菜分離得到的益生菌菌種在分離、鑑定與定性方面之相關文獻是很有限的，因此本研究目的為分離、鑑定與定性來自芥菜的益生菌菌種。透過數種檢測從菌種的生理與型態來鑑定與定性分離得到的益生菌種，例如革蘭氏染色、芽孢染色、過氧化氫酶試驗、硝酸鹽試驗和高效液相層析法等，API 50 CHL 和定序也可用於確認所分離得到的菌種類型。以耐低 pH 值、耐膽鹽、耐膽鹽水解、模擬人體胃腸道與溶血之耐受性和抗菌能力等體外試驗來篩選具有成為益生菌潛力的菌株。從芥菜分離得到的菌株為革蘭氏陽性菌、不產芽孢、過氧化氫酶試驗呈現陰性，硝酸鹽試驗呈現陰性的有十種分離株，有八株歸為同型發酵，而二株則為異型發酵。從 API50 CHL 和 16S rRNA 定序結果得知分離株 10 號相似於 Lactobacillus plantarum MCC 2156，分離株 3 和 4 號相似於 L. brevis JNB7，分離株 2 和 8 號相似於 L. plantarum IIX2。所有分離株都具有抗大腸桿菌、金黃色葡萄球菌、綠膿桿菌和枯草桿菌之抗菌 活性。所有分離株都表現出γ-溶血活性和生產 γ-氨基丁酸的能力，但只有 L. brevis 04這株分離株具有膽鹽水解酶的作用。L. plantarum 和 L. brevis能耐酸、耐膽鹽、胃蛋白酶和胰酶。這些結果證實芥菜含有乳酸菌(LAB) 種為 L. plantarum MCC 2156、 L. brevis JNB7 和 L. plantarum IIX2，由於 L. brevis在各種益生菌試驗結果良好故最具有成為益生菌潛力之菌株。
並列摘要
Sawi asin is one type of traditionally fermented non-dairy products and often consumed by Indonesian. Fermentation of Sawi asin is naturally fermentation, in contrast to fermented dairy products which do not occur naturally. This traditional fermented food is a plentiful source of microorganisms. However, isolation, identification and characterization probiotic from Sawi asin has limited investigation. The aim of this research was to isolate, identify, and characterize probiotic bacteria from Sawi asin. There are several tests to characterize and identify the isolates through physiology and morphology, such as Gram staining, endospores staining, catalase tests, nitrate tests and HPLC. API 50 CHL and sequencing also used to determine the type of isolates. In vitro test was used for screening potential probiotic bacteria that is resistance to low pH, resistance to bile salts, bile salts hydrolysis, tolerance to simulate human gastrointestinal tract, haemolytic and antimicrobial ability. Ten isolates obtained from Sawi asin are Gram-positive, non-spore forming, catalase negative and nitrate negative bacteria. Eight isolates classified as homofermentative, while 2 isolates as heterofermentative. From the results of API 50 CHL and 16S rRNA sequence, isolate 10 is similar to Lactobacillus plantarum MCC 2156, isolates 3 and 4 are similar to L. brevis JNB7 and isolates 2 and 8 are similar to L. plantarum IIX2. All of strain has antibacterial activity against Escherechia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Bacillus subtilis. All of the isolates exhibited γ–haemolytic activity and produced GABA, but only L. brevis strain 04 has bile salt hydrolase effect. L. plantarum and L. brevis could resistance to acid, tolerance to bile salts, pepsin and pancreatin. These results confirm that the Sawi asin contains lactid acid bacteria (LAB) that is L. plantarum MCC 2156, L. brevis JNB7 and L. plantarum IIX2, among that, Lactobacillus brevis is the best candidate of probiotic due to good results in probiotic evaluations.
參考文獻
延伸閱讀
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