|Abstract: ||摘 要 本研究是將市售蜈蚣藻 (Grateloupia filicina) 萃取多醣經α-amylase、cellulase、β-galactosidase、或由 Pseudomonas vesicularis MA103所產 agarase 等四種酵素水解後，挑出獲得寡單醣量較高的兩種酵素進行水解，以乳酸菌發酵該水解物，分析此乳酸發酵液之抗氧化能力。在相同酵素活性單位 (100 unit) 下水解蜈蚣藻多醣液，就水解後的還原醣量而言，cellulase 效果較佳，agarase 次之，且隨著酵素活性愈高，還原醣量愈高。 以 (A) 5% Lactobacillus (Lb.) rhamnosus BCRC14068、(B) 5% Enterococcus (Ent.) faecalis BCRC13076、與 (C) 2.5% Lb. rhamnosus BCRC14068 及 2.5% Ent. faecalis BCRC13076 於 37℃ 分別發酵 (1) 0.5% 蜈蚣藻多醣液經 agarase 及 cellulase (500 unit) 水解所得水解液及 (2) 該蜈蚣藻水解液添加 0.5% sucrose、1% beef extract 及 0.5% yeast extract (CN)，共得六組乳酸發酵液。結果顯示隨著發酵時間的增加，pH 值隨著下降。其中以 L-Gra-CN-FC5 組 Lb. rhamnosus BCRC14068 (2.5%) 及 Ent. faecalis BCRC13076 (2.5%) 發酵外加碳源與氮源的蜈蚣藻水解液反應至 9 hr 時，發酵液之 pH 值即可降至 4.6 以下，得一安全性產品，可滴定酸度為 0.74%，乳酸菌數 9.39 log CFU/mL。 六種乳酸發酵液在螯合亞鐵離子能力、血紅素催化亞麻油酸自氧化抑制率、還原力等抗氧化能力上，均以蜈蚣藻水解液另外添加碳氮源的基質發酵所得之發酵液較佳，除清除 DPPH 自由基能力。六種乳酸發酵液的可溶性多酚類含量，以 Lb. rhamnosus BCRC14068 及 Ent. faecalis BCRC13076 同時發酵外加碳源與氮源的蜈蚣藻水解液所含可溶性多酚類含量有 1,087.64 ± 0.31 GAE (gallic acid equivalents, μg/mL)，較其餘五種乳酸發酵液呈現較高多酚類含量 (400.27 ± 1.75 - 975.92 ± 4.90 GAE)。推測蜈蚣藻乳酸發酵液本身所含的胜肽、有機酸與可溶性多酚類等物質可能是提供抗氧化能力的成分。|
Abstract The polysaccharide extracts of Grateloupia (G.) filicina which was purchased from the market were digested by four enzymes, α-amylase, cellulase, β-galactosidase, and agarase that produced from Pseudomonas vesicularis MA103. From the four hydrolysates, two enzymes were selected by higher reducing sugar contents, and the polysaccharide extracts of G. filicina were digested by the two enzymes. The antioxidation of lactic acid fermented the hydrolysates was studied. The polysaccharide extracts of G. filicina were digested by the same activity unit (100 unit) of those four enzymes. The highest reducing sugar contents of those hydrolysates were digested by cellulase and agarase. The larger the activity units of enzyme were, the higher the reducing sugars contents were. Six lactic acid (LA) fermentation products were cultured in media (1) the hydrolysate of 0.5% aqueous solution of polysaccharide extracts of G. filicina digested by 500 unit agarase and cellulase or (2) this G. filicina hydrolysate added 0.5% sucrose, 1% beef extract, and 0.5% yeast extract.(CN) And these two media was each fermented by three groups lactic acid bacteria, group A: 5% Lb. rhamnosus BCRC14068, group B: 5% Ent. faecalis BCRC13076, group C: 2.5% Lb. rhamnosus BCRC14068 and 2.5% Ent. faecalis BCRC13076. The results of pH value of six LA fermentation products were decreasing as the time of fermentation increased. The pH value of LA fermentation products that cultured in the hydrolysate added carbon and nitrogen source (L-Gra-CN-FC5 ) achieved below 4.6 at fermentation time of 9 hr. That can be a secure product, and the titratable acidity is 0.74%, lactic acid bacteria count is 9.39 log CFU/mL Antioxidative properties, including ferrous ion chelating effect, inhibition of the hemoglobin-catalyzed peroxidation of linoleic acid, and reducing power of six LA fermentation products were better as the hydrolysate of G. filicina added carbon and nitrogen source than the hydrolysate of G. filicina, except for DPPH radical scavenging effect. The soluble total polyphenol contents of the hydrolysate of G. filicina added carbon and nitrogen source that fermented by Lb. rhamnosus BCRC14068 and Ent. faecalis BCRC13076 was recorded 1,087.64 ± 0.31 GAE (gallic acid equivalents, μg/mL) higher than other fermentation products. And the soluble total polyphenol contents of the other fermentation products were recorded from 400.27 ± 1.75 - 975.92 ± 4.90 GAE. The antioxidative constituents of LA fermented G. filicina may be peptides, organic acid, and soluble total polyphenol contents.