Studies on the Biological Activities and Food Application of the Hydrolysate Derived from Agarase Digested Algal Polysaccharides an Its Lactic Acid and Acetic Acid Ferment Product

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Project title

Code/計畫編號

NSC92-2313-B019-021

Translated Name/計畫中文名

海藻多醣經洋菜分解酵素水解物及其乳酸和醋酸發酵產物之生理活性與食品應用之探討

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=848627

Year

2003

Start date/計畫起

01-08-2003

Expected Completion/計畫迄

01-07-2004

Bugetid/研究經費

750千元

ResearchField/研究領域

食品科技(農)

本研究在將 Bacteriological (Agar No. 1) (Oxoid)、Agar powder, Bacteriological (HiMedia) 、 BactoTM agar (Difco) 、與 BitekTM agar (Difco) 等四種市售實驗級洋菜，分別簡稱為 Oxo、Him、Bac、及 Bit。光惠洋菜與惠光洋菜等購自台南市之二種食用級洋菜，分別簡稱為 Gua 及 Hui。石花菜、龍鬚菜、海菜、與紫菜等四種藻類萃取多醣，分別簡稱為 Gel、Gra、Mon、及 Por。此十種海藻多醣經 Aeromonas salmonicida MAEF108 及/或 Pseudomonas vesicularis MA103 所產 agarases 分解後，可得藻類寡醣混合物 (algal-oligosaccharide-lysates, AOLs)，然後分析抗氧化性、抗致突變性、抑菌性及所含可溶性多酚類含量。接著分別從十種海藻多醣，各挑一組經 agarases 分解所得 AOLs 具較高寡醣量者，以乳酸菌發酵藻類寡醣混合物，然後分析所得藻類寡醣混合物乳酸發酵液之抗氧化性、抗致突變性、及抑菌性。海藻多醣經 MAEF108 及/或 MA103 所產 agarases 分解後，可得 120 種 AOLs 。十種海藻多醣在還原力、清除 DPPH 自由基能力、螯合亞鐵離子能力、抑制血紅素催化亞麻油酸自氧化、過氧化氫清除效應、羥基自由基清除效應等六種抗氧化性測試中，皆以紫菜多醣較其餘九種海藻多醣呈現高的能力。而 120 種 AOLs 之還原力分佈於 51.8 ± 1.1 至 3.2 ± 0.8 ascorbic acid reducing power equivalent (AARPE) 間，以 A250-Por 為最高。而清除 DPPH 自由基能力介於 68.3 ± 0.7 至 0.4 ± 0.18%，以 108-Por500 為最佳。至於螯合亞鐵離子能力則在 93.1 ± 0.2 與 1.5 ± 0.2% 間，以 108-Por500 較其餘 AOLs 高。然後抑制血紅素催化亞麻油酸自氧化在 98.5 ± 1.0 到 5.0 ± 3.1% 間，並以 103-Gra500 最高的抑制率。接著清除過氧化氫有 35.9 ± 5.4 至 0.1 ± 0.2% 且以 108-Mon100 為最佳。最後羥基自由基則無清除效應。藻類寡醣混合物之抗致突變性方面，未添加 S9 時，60 種 AOLs 對 4NQO 所誘導 TA98 抑制率在 11.8 至 94.1% 間，並以 108-Oxo100 較其餘 AOLs 有較佳抑制率。而對 4NQO 所誘導 TA100 突變抑制率分佈在 3.5 至 97.7% 間，以 103-Bac500 、 103-Por500 、 108-Por100 及 B500-Bac 較其餘 AOLs 有較高抑制率。至於在添加 S9 時，60 種 AOLs 對 B[a]P 所誘導 TA98 之突變抑制率介於 17.8 至 89.0% 間，而 TA100 之突變抑制率則在 23.9 至 99.3% 間，此二項皆以 B500-Por 較其餘 AOLs 有較高抑制率。 120 種 AOLs 的可溶性多酚類含量在 70.4 ± 17.4 - 462.2 ± 1.6 gallic acid equivalents (GAE, g/mL) 間，且以 108-Por500 較其餘藻類寡醣混合物為佳。 AOLs 呈現出抗氧化性及抗致突變性與其所含可溶性多酚類含量為正相關。 AOLs 經 35 株乳酸菌篩選可利用發酵性中，以 Enterococcus faecalis BCRC13076 及 Lactobacillus plantarum BCRC14068 二株乳酸菌，在氮源為 beef extract 及 yeast extract 下，可使 pH 值下降至 4.6 以下。以 BCRC13076 及 / 或 BCRC14068 分別發酵 (1) 十種海藻多醣， (2) 十種海藻寡醣混合物，(3) 十種海藻寡醣混合物添加 0.5% 半乳糖後，共得 90 種乳酸發酵液，結果顯示隨著發酵時間的增加， pH 值隨著下降，當發酵至 18 小時後，十種海藻寡醣混合物之 pH 值可降至 4.6 以下，乳酸菌數則隨著發酵時間的增加而增加。 90 種乳酸發酵液的抗氧化性方面，除清除 DPPH 自由基能力外，其餘五種抗氧化性，皆隨著半乳糖量的增加而上升。還原力分佈於 10.3 ± 0.4 至 40.3 ± 0.2 AARPE 間。清除 DPPH 自由基能力在 79.6 ± 1.0 至 4.7 ± 2.5 %。亞鐵離子有 97.5 ± 0.7 至 16.2 ± 2.1% 的螯合能力。血紅素催化亞麻油酸自氧化則有 128.7 ± 1.3 至 3.2 ± 5.5% 抑制率。過氧化氫清除效應為 95.6 ± 0.6 至 37.3 ± 0.9%。羥基自由基清除能力分布於 71.5 ± 2.2 至 0.0 ± 0.0% 未添加 S9 時，90 種乳酸發酵液對 4NQO 所誘導 TA98 之突變抑制率分佈在 26 至 99% 間，對 TA100 之突變抑制率則分佈在 33 至 97% 間。在添加 S9 時，對 B[a]P 誘導 TA98 之突變抑制率分佈在 4 至 98% 間，對 TA100 之突變抑制率分佈在 33 至 99% 間。總之，龍鬚菜、海菜、與紫菜寡醣混合物，有被開發為具抗氧化及抗致突變等生理功能保健食品之潛力。 Four commercial and experimental grade quality agars, Bacteriological (Agar No. 1) (Oxoid), Agar powder, Bacteriological (HiMedia), BactoTM agar (Difco), BitekTM agar (Difco) as abbreviated as Oxo, Him, Bac, and Bit, respectively. Two food grade quality samples, Guanghui agar and Huiguang agar, were purchased from Tainan city and abbreviated as Gua, and Hui, respectively. The algal polysaccharide extracts (APEs) of Gelidium spp., Gracilaria spp., Monostroma nitidum, and Porphyra dentate were abbreviated as Gel, Gra, Mon, and Por, respectively. These ten seaweed polysaccharides were digested by agarases that produced from Aeromonas salmonicida MAEF108 and/or Pseudomonas vesicularis MA103. The various products obtained from those ten polyasccharides digested by agarases were designated as algal-oligosaccharide-lysates, (AOLs) and then the analyses of their antioxidative properties, antimutagenicities, antimicrobial properties, and soluble total polyphenol (TP) contents were conducted. The each of highest reducing sugar of AOL from ten seaweed polysaccharides were selected and preceded fermentation by lactic acid bacteria. The AOLs fermentation products by lactic acid bacteria were evaluated antioxidative properties, antimutagenicity, and antimicrobial properties. 120 AOLs were obtained from ten polyasccharides digested by agarases that produced from MAEF108 and/or MA103. In antioxidative properties of ten seaweed polysaccharides, Por show higher antioxidative properties in reducing power, DPPH radical scavenging effect, ferrous ion chelating effect, inhibition of the hemoglobin-catalyzed peroxidation of linoleic acid, hydrogen peroxide scavenging effect, and hydroxyl raducals scavenging effect than others seaweed polysaccharides. The reducing power of 120 AOLs were ranged from 51.8 .plmin. 1.1 to 3.2 .plmin. 0.8 ascorbic acid reducing power equivalent (AARPE) and A250-Por richer than others AOLs. DPPH radical scavenging effect of 120 AOLs were recorded from 68.3 .plmin. 0.7 to 0.4 .plmin. 0.18% and 108-Por500 higher than others AOLs. Ferrous ion chelating effect of 120 AOLs were displayed from 93.1 .plmin. 0.2 to 1.5 .plmin. 0.2% and 108-Por500 richer than others AOLs. In inhibition of the hemoglobin-catalyzed peroxidation of linoleic acid, the inhibition were 98.5 .plmin. 1.0 to 5.0 .plmin. 3.1% and 103-Gra500 higher than others AOLs. The scavenging were ranged from 35.9 .plmin. 5.4 to 0.1 .plmin. 0.2% in hydrogen peroxide scavenging effect and 108-Mon100 richer than others AOLs. As for hydroxyl raducals scavenging effect had no any result of 120 AOLs. The antimutagenicity of 60 AOLs against 4NQO to S. typhimurium TA98 were 11.8 to 94.1% and 108-Oxo100 higher than others AOLs. The antimutagenicity of 60 AOLs against 4NQO to S. typhimurium TA100 were among 3.5 to 97.7% and 103-Bac500, 103-Por500, 108-Por100 and B500-Bac richer than others AOLs. In S9 mixtrues, the antimutagenicity of 60 AOLs against B[a]P to S. typhimurium TA98 were reveal 17.8 to 89.0%. The antimutagenicity of 60 AOLs against B[a]P to S. typhimurium TA100 were 23.9 to 99.3%. And B500-Por was higher than others AOLs in later two tests. The soluble TP contents of 108-Por500 was lager than other AOLs and recorded from 70.4 .plmin. 17.4 - 462.2 .plmin. 1.6 gallic acid equivalents (GAE, g/mL) of 120 AOLs. The antuoxidative properties and antimutagenicity of AOLs were correlate to their soluble TP contents and reveal positive correlation. To select the AOLs that could be metabolized by 35 lactic acid bacterial strains, the results showed that two lactic acid bacterial strains, which were Enterococcus faecalis BCRC13076 and Lactobacillus plantarum BCRC14068 were could use AOLs with beef extract and yeast extract were added as nitrogen source. And the pH values of fermentation products could decrease to below 4.6. 90 lactic acid .....

Keyword(s)

洋菜
洋菜脢
石花菜
龍鬚菜
海菜
紫菜
藻類寡醣混合物
抗氧化
抗致突變
乳酸發酵
Agar
Agarases
Gelidium spp
Gracilaria spp
Monostroma nitidum
Porphyra dentate
Algal-oligosaccharide-lysates (AOLs)
Antioxidative properties
Antimutagenic activity
Lactic acid fermentation

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Studies on the Biological Activities and Food Application of the Hydrolysate Derived from Agarase Digested Algal Polysaccharides an Its Lactic Acid and Acetic Acid Ferment Product

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