The Studies on the Purification and Composition Analysis as Well as the Application on Food of Extra(III)

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Project title

Code/計畫編號

NSC91-2313-B019-006

Translated Name/計畫中文名

海洋附著細菌胞外黏性物質之純化和組成分析以及在食品上應用之探討(III)

Project Coordinator/計畫主持人

Chorng-Liang Pan

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=737323

Year

2002

Start date/計畫起

01-08-2002

Expected Completion/計畫迄

01-07-2003

Bugetid/研究經費

877千元

ResearchField/研究領域

化學工程
食品科技(工)

本研究在測試海洋附著細菌產生較多細胞外黏性物質 (Extracellular adhesive substances, EAS) 之最適培養基及培養條件之生產條件，並以最適培養基與最適培養條件培養海洋附著細菌將其胞外黏性物質萃取分離與純化，再將 EAS 萃取物做為抗氧化劑的可行性與評估做為食品添加物之安全試驗。菌株 Micrococcus sp. strain DB05B 為分離自附著於浸漬海水中材料片之海洋黏性菌株，由實驗結果顯示於 26℃、 150 rpm 振盪培養 48 小時下，當以 5% 乳糖與 0.71% yeast extract 取代黏液篩選基礎培養基 (mucous screening basal broth, MSBB) 中之碳源與氮源時，可使胞外黏性物質 (extracellular adhesive substance, EAS) 的產量由 0.29-0.31 g/L 分別提高至 0.74 g/L 與 0.65 g/L。當添加 10 ppm 的 Mn+2 離子與 1.5 mM disodium-β-glycerophosphate (DSGP) 於 MSBB 中，分別可獲得較高的 EAS 產量 (0.57 與 0.68 g/L)。在培養條件方面，經測試在不同起始 pH 值、培養溫度、振盪培養轉速後，可得到最適生產 EAS 之個別條件為：起始 pH 值為 7.2、培養溫度為 26℃、振盪培養轉速為 150 rpm，其 EAS 產量可提升至 0.80-0.85 g/L。將最佳生產條件與培養基組成綜合成為菌株 DB05B 之乳糖修飾黏液產生培養液 (modified-mucous production broth-lactose, M-MPB-L)。在發酵槽中以 100 rpm 培養菌株 DB05B 之攪拌速率可得更佳之 EAS 產率 (1.05 ± 0.02 g/L)。菌株 SCA38 於 MSBB 、 M-MPB-M 、及 M-MPB-G 三種培養液中，所得之 EAS，依 10-50 mg/mL 三種濃度；菌株 DB05B 於 MSBB 和 M-MPB-L 培養液中所得之 EAS，以 5、10、15 mg/mL 三種濃度測試其抗氧化能力。在清除 DPPH 自由基能力方面，二株菌經測試結果均未呈現顯著效果，菌株 SCA38 以 M-MPB-M 培養所得 EAS 對於抑制血紅素催化亞麻油酸自氧化能力較為顯著，在濃度為 10 mg/mL 時，可達到 93.04 ± 0.58%，以 MSBB 與 M-MPB-G 培養所得 EAS 則隨著測試濃度之增加而其抑制血紅素催化亞麻油酸自氧化能力亦隨之上升 (5.51%-79.81%)；在菌株 DB05B 方面，以 MSBB 培養所得 EAS 較 M-MPB-L 為佳。抗致突變能力方面，菌株 SCA38 以 MSBB 所產 EAS 於濃度 5.0% 時，對 MMNG 及 B[a]P 抑制率分別為 37.1% 與 5.5%，濃度在 1.0-5.0% 之 M-MPB-G 所產 EAS 之抑制 MMNG 能力在 85.7-88.6% 之間， B[a]P 抑制率為 33.7-34.4% 範圍間。 The investigation was conducted to examine the optimal conditions and best ingredient coombination of incubation medium for the extracellular adhesive substances (EAS) producing marine biofouling bacteria. Not only evaluate the feasibility of EAS to be an antioxidant, but also the food safety of these marine bacteria EAS was examined. Micrococcus sp. strain DB05B was isolated from the biofouling material suspended in sea water. While 5% lactose and 0.71% yeast extract were used to replacing the original carbon source and nitrogen source of mucous screening basal broth (MSBB), the results indicated that the extracellular adhesive substance (EAS) yields were increased from 0.29-0.31 g/L up to 0.74 g/L and 0.65 g/L, respectively. Ten ppm Mn2+ metal or 1.5 mM disodium-.beta.-glycerophosphate (DSGP) was added into the MSBB individually, and then higher EAS yields, 0.57 and 0.68 g/L were achieved, respectively. Modified-mucous production broth-lactose (M-MPB-L) is composed of the individual best cultured medium ingredient for strain DB05B. To test the different agitating speed on the EAS yield for strain DB05B, the higher EAS yield of this strain were observed while 100 rpm was employed. The EAS recovered from the cultivation of strain SCA38 in MSBB M-MPB-M, and M-MPB-G; strain DB05B in MSBB and M-MPB-L, were used to test their capability on antioxidation in the concentration ranged from 10 to 50 mg/mL and 5 to 15, individually. Both the antioxidative performance of those EASs were not remarkable on the scavenging DPPH free radical. As to the EAS recovered from the strain SCA38 cultured MSBB and M-MPB-G, the inhibition performances on the hemoglobin catalyzing linoleic acid were ranged from 5.51% to 79.81% as the added concentration of EAS was increased. The strain DB05B lyophilized EAS derived from MSBB showed a better result than the EAS derived from M-MPB-L. As to the antimuta-genicity of EAS (5%) that produced from strain SCA38 cultured in MSBB, the inhibition rate to the mutation caused by MMNG or B[a]P could be 37.1% and 5.5%, respectively. In the same testing system, the EAS of strain SCA38 that derived from M-MPB-G had inhibition rates while against the mutation caused by MMNG were among 85.7-88.6%.

Keyword(s)

細胞外黏性物質
海洋附著細菌
發酵槽生產
抗氧化性
抗致突變性
Extracelullar adhesive substance (EAS)
Marine biofouling bacteria
Fermenter production
Antioxidation
Antimutagenicity

DSpace CRIS

The Studies on the Purification and Composition Analysis as Well as the Application on Food of Extra(III)

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