The Studies on the Purification and Composition Analysis as Well as the Application on Food of Extracellular Adhesive Substance Produced by Marine Biofouling Bacteria (II)

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Project title

Code/計畫編號

NSC90-2313-B019-039

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(II)

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=648876

Year

2001

Start date/計畫起

01-08-2001

Expected Completion/計畫迄

01-07-2002

Bugetid/研究經費

694千元

ResearchField/研究領域

食品科技(農)

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，並以上述組合成分與條件培養海洋附著細菌以取得最大量的胞外黏性物質產物，將所得EAS經萃取、分離與初步純化，並進行組成分析與初步應用分析。Neisseria sp. strain SCA38培養在添加1 mg MnCl/sub 2/．7H/sub 2/O之MSBB中時，其EAS之產率是四組金屬離子測試中最高者，可達到0.13g/L，RV為1.04。Strain SCA38培養在添加1.5 mM disodium-β-glycerophosphate (DSGP)所獲得之EAS產量較1.0或1.5 mM DSGP、K/sub 2/HPO/sub 4/、KH/sub 2/PO/sub 4/ (0.5-1.5 mM)高。當Strain SCA38使用有機氮源成分如Yeast extract與Polypeptone/yeast extract時，EAS產量及菌液的相對黏度均較以無機氮源時為高。在MSBB培養液中置換不同碳源，以26℃並150 rpm振盪培養Strain SCA38，結果顯示Strain SCA38所產之聚集胞外黏性物質在含有5% Mannose之培養基培養44 hr後，培養基中可得到0.81 g/L之EAS產量。在培養條件方面，Strain SCA38於26℃下培養，可得到較高的EAS產量(0.77 g/L)。Strain SCA38培養在起始pH值為8.2之MSBB培養液時，可產生最高之EAS產量(0.71 g/L)。Strain SCA38，以150 rpm振盪培養時，所得RV 1.07與EAS產量0.83 g/L，均較以50與100 rpm之振盪速度培養時為高。將測試出之最適培養基添加六種不同材質，活性碳片、幾丁質片、小木塊、小鋼珠、聚丙烯珠及玻璃珠並於培養溫度26℃、振盪轉速150 rpm觀察其對Strain SCA38與Strain SDC07 EAS產量所造成的影響。實驗結果指出Strain SCA38與Strain SDC07在添加幾丁質片所得產量最高分別為0.98±0.27 g/L與1.03±0.27 g/L，活性碳片次之，分別為0.72±0.10與0.98±0.14 g/L。RV值之表現仍以添加幾丁質片者表現較佳，如Strain SCA38為1.14±0.02與Strain SDC07 1.52±0.53。利用發酵槽來生產Strain SDC07胞外黏性物質，結果顯示Strain SDC07在26℃、150 rpm、通氣量2 L/min，控制pH值5.20的狀態下，獲得EAS產率最高(0.49-1.03 g/L)。以5.0 L發酵槽通氣攪拌培養Strain SCA38，觀察不同攪拌速率對Strain SCA38之EAS生產量的影響，實驗結果顯示RV、好氣性生菌數、及EAS產量均以攪拌速率200 rpm進行培養時所得結果較佳，Strain SCA38之EAS在控制pH值8.20的條件下可獲得較大EAS產量(1.33 g/L)。菌株SDC07所產EAS在Distilled water、1.0-5.0% NaCl、80.0% formic acid及99.0% Dimethyl sulphoxide中可完全溶解，而菌株SCA38所產EAS在去離子水、1.0-4.0% NaCl及Formic acid(溶解度為85.6-96.4%)中可以完全溶解。不同濃度之Strain SDC07與Strain SCA38的EAS水溶液之相對黏度含隨著EAS濃度增加而增加。相同濃度之Strain SDC07、Strain SCA38、Strain DB05B與Strain SDC01的EAS水溶液，會隨著溶液中NaCl濃度增加其黏度表現有減緩的趨勢。在探討多醣類對大豆分離蛋白質乳化特性的影響，結果顯示在0.2% Strain SDC07之EAS的濃度時，EAS之乳化活性達到最高(70.0%)，而Strain SCA38則是在濃度0.8% EAS的濃度時，其乳化活性達到最高(48%)。Strain DB05B在濃度1.0% BAS其乳化活性達到最高(42%)。0.2%之Strain SDC01的EAS濃度時，EAS之乳化活性最高(78.57%)。在0.6% Strain SDC07之EAS的濃度時，乳化活性最低(53.0%)，之後趨於平緩。將乳化物置於80℃中加熱30 min以評估在不同濃度的Strain SDC07之EAS添加下之乳化安定性，結果顯示EAS之乳化安定性皆呈現穩定的狀態(60.0-64.0%)，Strain DB05B與Strain SDC01 EAS之 This investigation was conducted to obtaining the optimal compositions of incubation medium and incubation conditions employed on the culturing the extracellular adhesive substances (EAS) producing marine biofouling bacteria. Four metal ions or three buffer agents was added individually into the MSBB to testing the EAS production of Neisseria sp. strain SCA38. The higher EAS productions of this strain were also observed while 1.5 mM Disodium-.beta.-glycerophosphate (DSGP) or 1.0 mg/L MnCl/sub 2/.bul.7H/sub 2/O was employed in the modified medium. And as yeast extract or polypeptone/yeast extract were used to take place the original nitrogen source, the EAS yield of strain SCA38 were higher than the inorganic nitrogen replacement groups. Five monosaccharide and three disaccharides were used individually to replace the carbon source of MSBB, the results indicated that the substitution of mannose in the MSBB performed higher EAS yield (0.81 g/L) than others. In the study of incubation conditions for the maximum EAS production of strain SCA38, the results showed that while the initial pH at 8.2, shaking speed with 150 rpm, or incubation temperature at 26.degree.C, the higher EAS yields ranged from 0.71 to 0.83 g/L were observed. Marine biofouling bacterium strain SDC01 and SCA38 was cultured in the M-MPB with the addition of substratum such as actived carbon flake (ACF), chitin flake, wood piece, stainless steel beads, glass beads, or polypropylene beads and incubation temperature at 26.degree.C, shaking speed with 150 rpm. The EAS production results indicated that the chitin flake added to M-MPB performed the highest EAS yield as 1.03 and 0.98.plmin.0.27 g/L and 1.03.plmin.0.27 g/L for strain SDC01 and SCA38, respectively. Followed by the ACF group performed the EAS yield as 0.72.plmin.0.10 and 0.98.plmin.0.14 for strain SDC01 and SCA38 receptively. As to the RV, results the chitin addition group reached a better as 1.14.plmin.0.02, 1.52.plmin.0.53 for strain SCA38 and strain SDC07, respectively. The strain SDC07 being cultured in broth uncontrolled pH or controlled at pH 5.2 employed in a 5.0 L fermenter, with incubation temperature at 26.degree.C, shaking speed 150 rpm, and 2 L/min aeration rate. The result showed that the higher EAS yield raised to 0.49-1.03 g/L in the controlled pH incubation group. To test the different agitating speed on the EAS yield for strain SCA38, the higher EAS yield of this strain were observed while 200 rpm was employed in the incubation condition. In addition, the higher yield (1.33 g/L) of EAS produced by strain SCA38 could be achieved, while this strain was incubated under constant pH at 8.20 in a 3.0 fermenting broth (aeration: 2 L/min, 26.degree.C; and 150 rpm). The solubility of rehydrated aqueous solution with strain SDC07 EAS powder was dissolved distilled water, 1.0-5.0% NaCl, 80% formic acid, and 99% dimethyl sulphoxide. For strain SCA38 the dehydrated aqueous solution with EAS was dissolved distilled water, 1.0-4.0% was dissolved distilled water. The RVs of EAS rehydrated aqueous solutions derived from strains SDC07, SCA38, DB05B, and SDCO1 were increased when the concentration of each EAS was increasing in the reacting solution. Effect of EAS addition from strain SDC07 and SCA38 on emulsifying activity and emulsion stability while mixed with isolated soy protein were examined, the result showed that emulsifying activity 0.2% EAS (strain SDC07) performing the highest EA (70.0%). For strain SCA38, 0

Keyword(s)

海洋附著細菌
胞外黏性物質
純化
成分分析
食品
Marine biofouling bacteria
Extracellular adhesive substance
Purification
Component analysis
Food

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The Studies on the Purification and Composition Analysis as Well as the Application on Food of Extracellular Adhesive Substance Produced by Marine Biofouling Bacteria (II)

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