台灣酸菜中高胞外多醣益生乳酸菌之篩選與其醃漬液製作酸菜醃蛋可行性之研究

黃靖堯

doi:10.6845/NCHU.2007.00418

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台灣酸菜中高胞外多醣益生乳酸菌之篩選與其醃漬液製作酸菜醃蛋可行性之研究

Isolation of Exopolysaccharide-Producing Probiotic Lactic Acid Bacteria from Mustard Pickle in Taiwan and Feasibility of Making Salted Duck Eggs Using Mustard Pickle Brine

黃靖堯 , Masters　　Advisor：方繼　　

繁體中文 DOI： 10.6845/NCHU.2007.00418

酸菜；乳酸菌；胞外多醣；益生菌；酸菜醃蛋；鹹蛋； mustard pickle ； lactic acid bacteria ； exopolysaccharide ； probiotic ； mustard pickled egg ； salted egg

Abstract │ Reference (186) │ Times Cited (4) │ Altmetrics

Abstract 〈TOP〉

本研究以酸菜為實驗主題，分別對酸菜與酸菜醃漬液進行研究，並分成兩個章節：第一章為台灣酸菜中高胞外多醣益生乳酸菌之篩選；第二章為酸菜醃漬液製作酸菜醃蛋可行性之研究。

第一章：本實驗收集台灣北部(新竹縣北埔鄉)、中部(雲林縣大埤鄉)及南部地區(高雄縣美濃鎮)之酸菜樣品，從中篩選具有胞外多醣(exopolysaccharide, EPS)產生能力的乳酸菌，除進行菌種鑑定外並評估其益生菌條件，以及探討葡萄糖濃度對EPS生產的影響。由酸菜樣品中分離出單一菌落，該分離菌中具有產酸能力、觸媒試驗負反應、革蘭氏染色正反應、鏡檢為桿菌或球菌之乳酸菌特性者，共計39株。進一步分析其EPS產量，以高雄縣美濃鎮的酸菜樣品所獲得之乳酸菌SLC 13可產生最多的EPS，產量可達到400 mg/L以上。而在耐酸試驗、耐膽鹽試驗、模擬消化道試驗、抑菌試驗及抗生素敏感試驗，乳酸菌SLC 13皆優於其它篩選出之乳酸菌，具有益生菌之條件。另外，以API 50 CHL及16S rDNA序列進行菌種鑑定，其結果皆顯示乳酸菌SLC 13為Lactobacillus plantarum。以MRS broth為培養基，額外添加葡萄糖，發現MRS broth額外添加6 g/100 mL之葡萄糖，其EPS即可達到最大產量，為2.03 ± 0.05 g/L，菌數為8.88 ± 0.05 log10 CFU/mL，pH為3.54 ± 0.01，殘醣量為0.53 ± 0.01 g/L。本實驗成功於台灣酸菜樣品篩選出可產高胞外多醣並同時具有益生菌條件之乳酸菌。

第二章：本實驗以「雲林縣大埤鄉酸菜專業區」統一集中之酸菜醃漬液，與其醃漬液經蒸發結晶器回收之酸菜鹽，調和成飽和酸菜醃漬液來進行新產品醃漬鹹鴨蛋之開發。酸菜醃漬液的成份分析顯示，其鹽度為18.84 ± 0.23 %，pH值為4.96 ± 0.03，酸度相當於0.54 ± 0.04 g乳酸/ 100 g-酸菜醃漬液。酸菜醃漬液以酸菜鹽調合成飽和鹽溶液，再以市售鴨蛋進行醃漬，同時以飽和食鹽水做為對照組，分析鴨蛋醃漬期間蛋白與蛋黃的變化。醃漬期間酸菜醃蛋之蛋重有稍微下降，而對照組鹹蛋則有上升的趨勢。在蛋黃部份，酸菜醃蛋與對照組有相同的趨勢，蛋黃係數上升、鹽度上升、水分下降、總固形物上升及質地分析之硬度上升。兩組蛋白之蛋白係數及pH值皆下降，鹽度及總固形物皆上升，流變性質之G’、G’’皆下降和tan δ上升及η*下降。在官能品評方面採用九分制評分標準，酸菜醃蛋之蛋黃與蛋白，在外觀、顏色和質地方面，皆比市售鹹蛋與對照組鹹蛋得到皆得到較高之分數，而整體喜好上也有很好的接受性。本研究之實驗結果顯示，以酸菜醃漬液及酸菜鹽進行鴨蛋的醃漬，可開發出具酸菜風味的鹹鴨蛋，可做為雲林縣大埤鄉的地方特產之ㄧ，並達到酸菜醃漬液及酸菜鹽之再利用。

Parallel Abstract 〈TOP〉

The theme of this study is mustard pickle. We investigate mustard pickle and mustard pickle brine respectively. The study can divide into two chapter: the first chapter is “Isolation of Exopolysaccharide-Producing Probiotic Lactic Acid Bacteria from Mustard Pickle in Taiwan ” and the second chapter is “ Practicability of Making Salted Duck Eggs Using Mustard Pickle Brine ”.

The first chapter: this experiment collected mustard pickle form north (Beipu Township, Hsin Chu County), midland (Tapi Township, Yunlin County) and south (Mei Nong Township, Kaohsiung County) in Taiwan to isolated exopolysaccharide (EPS)-producing lactic acid bacteria (LAB). We evaluated probiotics property and EPS production related to media of different glucose concentration and then identifies the LAB. Based on initial characterization including acid production (positive), catalase (negative), Gram-stain (positive) and cocci or rods shape, thirty nine strains were isolated from mustard pickle. By evaluating their EPS production, strain SLC 13, which was isolated from mustard pickle of Mei Nong Township Kaohsiung County, produced the maximum EPS (more than 400 mg/L) among these isolates. Through acid tolerance test, bile tolerance test, imitation of digestional tract condition test, antibacterial test and antibiotic susceptibility test, we found that LAB SLC 13 performed better than the other isolates and has the potential to be a probiotic. Further identification of LAB SLC 13 was performed by using API 50 CHL and 16S rDNA, results showed that LAB SLC 13 is Lactobacillus plantarum with more than 99 % confidence. By culturing L. plantarum SLC13 in MRS broth/ 100 ml with 6 g glucose, high EPS production (EPS 2.03 ± 0.05 g/L) and good growth (viable counts 8.88 ± 0.05 log10 CFU/mL) was observed. The finial pH and residual sugar was found to be 3.54 ± 0.01 and 0.53 ± 0.01 g/L, respectively. The experiment isolated EPS-producing and probiotics lactic acid bacteria successfully from mustard pickle in Taiwan.

The second chapter: mustard pickle brine with NaCl content 18.84 ± 0.23 %, pH 4.96 ± 0.03, and total acidity level of lactic acid of 0.54 ± 0.04 %, was from the area for making mustard pickle in Tapi Township Yunlin County in Taiwan. Mustard pickle salt is made from mustard pickle brine by evaporating crystallizer. We used both mustard pickle brine and mustard pickle salt for preparing saturated brine to make salted duck eggs. Salted duck eggs making from saturated salt brine preparing from commercial salt were used as control. The changes of egg yolk and egg white during pickling time were analyzed. The weight of mustard pickled eggs was slightly decreased during pickling time; however, weight increase was observed in control group. For egg yolk, increase of the parameters such as yolk index, NaCl content, total solids and hardness were observed during pickling time with moisture content as the exception. For both experimental and control groups, decrease in parameters for egg white including albumen index, pH, G’, G’’ and η* were found. On the contrary, increase in NaCl content, total solids and tan δ were observed. In sensory evaluation using nine point hedonic scales, results showed highest scores of mustard pickled eggs in appearance, color and texture in both egg yolk and egg white were obtained. Overall acceptance also gained higher scores. Our results showed that mustard pickle brine can make salted duck eggs with mustard pickle flavor and these salted eggs can be a speciality of Tapi Township Yunlin County or anywhere that produce mustard pickles. This experiment also indicated the possibility of reusing mustard pickle brine and mustard pickle salt by developing new salted egg products.

Reference ( 186 ) 〈TOP〉

行政院衛生署。2000。國民飲食指標。
連結：
陳智強。2004。培養條件對乳酸菌胞外多醣生產及抗氧化性之影響。台灣大學食品科技所碩士論文。
連結：
Adams, M. R. 1999. Safety of industrial lactic acid bacteria. J. Biotechnol. 68: 171-178.
連結：
Ahn, C., Thompson, D.C., Duncan, C., and Stiles, M.E., 1992. Mobilization and location of the genetic determinant of chloramphenicol resistance from Lactobacillus plantarum ca TC2R. Plasmid. 27: 169-176.
連結：
AOAC. 1984. “Official Methods of Analysis,” 14th Ed. Association of Official Analytical Chemists. USA: Washington, D. C.
連結：

Times Cited (4) 〈TOP〉

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