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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53733

Title: 探討綠藻石蓴做為營養補充物應用於乳酸發酵
The use of green macroalgae Ulva sp. as nutrient supplementation in lactic acid fermentation
Authors: Hung, Chin-Chieh
洪勤絜
Contributors: NTOU:Department of Food Science
國立臺灣海洋大學:食品科學系
Keywords: 綠藻石蓴;嗜酸乳酸桿菌;營養補充物;酵素水解;乳酸發酵
Ulva sp.;Lactobacillus acidophilus;Nutrient supplementation;Enzymatic hydrolysis;Lactic acid fermentation
Date: 2019
Issue Date: 2020-07-03T08:43:17Z
Abstract: 乳酸不僅為生物可分解材料─聚乳酸的主要原料,在製藥、食品、造紙和塑膠等工業上亦有廣泛的應用;微生物發酵為乳酸主要的生產方式,而大型藻類─綠藻石蓴中富含多醣與蛋白質,具有做為乳酸發酵基質之潛力。本研究利用乳酸菌發酵綠藻石蓴水解液並探討其乳酸產量。固定 109 CFU/mL 的 Lactobacillus rhamnosus BCRC 10940 與 BCRC 14068 於聚乳酸微管束陣列膜 (Poly-L-Lactic Acid Microtube Array Membrane, PLLA-MTAM),可分別得到 84.0 ± 6.4% 與 83.1 ± 5.0% 之包覆率,以此固定化菌體發酵含有 3% (w/v) 葡萄糖的 MRS 培養基 72 h 後之乳酸濃度分別為 15.32 ± 0.13 g/L 與 15.33 ± 0.19 g/L、乳酸轉換率分別為 0.55 ± 0.01 g/g sugar 與 0.58 ± 0.01 g/g sugar;另外也會產生非目標產物乙醇 5.25 ± 0.08 g/L 與 5.28 ± 0.02 g/L,而乙醇轉換率分別為 0.20 ± 0.00 g/g sugar 與 0.20 ± 0.00 g/g sugar,產物莫耳數比則為分別 1 : 0.70 與 1 : 0.70 (乳酸 : 乙醇)。透過酸水解及電透析法製備石蓴酸水解液,其葡萄糖含量為 9.35 ± 0.07g/L、鼠李糖含量為 39.27 ± 0.12 g/L,並能將其鹽度降至 8% (w/w) 以下。以 Lb. rhamnosus BCRC 10940 及 BCRC 14068 發酵 4% (w/v) 鼠李糖作為替代碳源之 MRS 培養基,Lb. rhamnosus BCRC 10940 雖能利用鼠李糖產生乳酸,最終乳酸濃度為 8.79 ± 0.04 g/L,但其轉換率為 0.36 ± 0.11 g/g sugar,最高乳酸生產速率達 0.28 ± 0.01 g/L∙h;而 Lb. rhamnosus BCRC 14068 則幾乎無法利用鼠李糖產生乳酸,僅消耗 0.94 ± 0.04 g/L 的鼠李糖,產生 0.46 ± 0.01 g/L 的乳酸。上述之實驗結果說明綠藻石蓴並不適合做為 Lb. rhamnosus BCRC 10940 與 BCRC 14068 之乳酸發酵基質中的碳源,而此二菌株也不適合做為發酵生產高濃度乳酸的菌株。利用 12% (w/v) 石蓴藻粉、纖維素酶 Cellulase 15 U/mL與蛋白酶 Umamizyme 5.40 U/g Ulva sp.、水解時間 48 h 的條件製備石蓴酵素水解液,其游離胺基酸含量為 103.06 ± 1.38 mM,葡萄糖與鼠李糖含量分別為 8.20 ± 0.12 與 6.33 ± 0.06 g/L。將石蓴酵素水解液之初始葡萄糖濃度調整至約 40 g/L 並添加 0.26 M 乙酸鈉作為緩衝劑,接種 Lb. acidophilus BCRC 10695 進行乳酸發酵,其最終乳酸濃度為 24.32 ± 0.40 g/L,最高乳酸生產速率為 0.58 ± 0.04 g/L∙h,乳酸轉換率為 0.86 ± 0.02 g/g sugar,最高乳酸菌數為 9.31 ± 0.03 Log CFU/mL。此結果表示 Lb. acidophilus 於石蓴酵素水解液中的生長狀況良好,且綠藻石蓴確實可以做為良好的營養補充物,替代乳酸發酵基質中的氮源提供游離胺基酸。
Lactic acid is not only the main raw material of polylactic acid, but also widely used in industry such as pharmaceutical, food, and plastics industries. Green alga Ulva sp., which contains high amount of polysaccharides and proteins, has the potential to be an alternative substrate for lactic acid fermentation. This study intends to use green alga Ulva sp. for lactic acid fermentation. The encapsulation efficiency of immobilized Lactobacillus rhamnosus BCRC 10940 and BCRC 14068 were 84.0 ± 6.4% and 83.1 ± 5.0%, respectively, by using Poly-L-Lactic Acid Microtube Array Membrane (PLLA-MTAM). The yields of lactic acid in fermentation of MRS broth using immobilized Lb. rhamnosus BCRC 10940 and BCRC 14068 were 0.55 ± 0.01 and 0.58 ± 0.01 g/g sugar, respectively. In addition, by-product ethanol was also producted, the yields of ethanol were 0.20 ± 0.00 and 0.20 ± 0.00 g/g sugar, respectively. The Ulva sp. acid hydrolysate was prepared by acid hydrolysis and electrodialysis, in which contained 9.35 ± 0.07g/L of glucose, 39.27 ± 0.12 g/L of rhamnose. The MRS broth with rhamnose as carbon source was fermented by Lb. rhamnosus BCRC 10940 and BCRC 14068, Lb. rhamnosus BCRC 10940 was able to use rhamnose and produce lactic acid, the yields of lactic acid were 0.36 ± 0.11 g/g sugar, and the highest lactic acid production rate was 0.28 ± 0.01 g/L∙h; Lb. rhamnosus BCRC 14068 was unable to produce lactic acid from rhamnose. The results above indicated that Ulva sp. is not suitable as carbon source in the lactic acid fermentation using Lb. rhamnosus BCRC 10940 and BCRC 14068. Lb. rhamnosus BCRC 10940 and BCRC 14068 are also not suitable as strains for the fermentation aims high production of lactic acid. 12% (w/v) Ulva sp. powder, 15 U Cellulase/mL, 5.40 U Umamizyme/g Ulva sp., and hydrolysis for 48 hours were used to prepare the Ulva sp. enzymatic hydrolysate, and the free amino acid content was 103.06 ± 1.38 mM. The Ulva sp. enzymatic hydrolysate was fermented by Lb. acidophilus, the highest lactic acid production rate was 0.58 ± 0.04 g/L∙h, the yield of lactic acid was 0.86 ± 0.02 g/g sugar, and the highest count of viable Lb. acidophilus cells was 9.31 ± 0.03 Log CFU/mL. These results indicated that Lb. acidophilus grew well in the Ulva sp. enzymatic hydrolysate, and Ulva sp. could be used as a good nutrient supplementation for lactic acid fermentation, to replace the nitrogen source in the media and provide amino acids.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010632045.id
http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53733
Appears in Collections:[食品科學系] 博碩士論文

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