酵母菌發酵穀類對乳汁分泌刺激之研究

Stimulation of Breast Milk Secretion by Yeast Fermented Cereal

吳芝妤(Jhih-Yu Wu)

指導教授 ： 蔣丙煌

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2014年)

https://doi.org/10.6342/NTU.2014.01835

摘要

母乳對於嬰兒的成長扮演很重要的角色，泌乳滿足下一代的營養需求，也為畜牧業帶來經濟價值。因此不管是母親或是畜牧業者，皆期望能提高泌乳量及其營養價值，尤其不希望有乳量缺乏的情況發生。傳統食品甜酒釀乃以根黴菌、酵母菌發酵食米製成，據說有增加泌乳量的效果，因此母親在做月子的時候，常常將其當作甜品飲用。而在畜牧業，則是有將酵母菌添加到飼料，或是直接餵食菌體來提升乳產量以及其營養價值之做法，但過去活體試驗中，大多推論酵母菌發酵產物或是直接攝食酵母菌會調整瘤胃菌相，以達到更高的消化吸收率，增進泌乳。本研究欲更進一步探討，酵母菌或其他微生物發酵產物是否會透過乳腺上皮細胞，直接影響泌乳，並且了解發酵產物的安全性。為了能夠快速篩選出有效且無毒性的發酵產品，因此本研究以山羊乳腺上皮細胞作為體外模式，進行細胞毒性試驗、以及beta-casein表現的試驗。由於固態發酵產物較液態發酵所需後續乾燥處理等步驟較符合經濟成本，本研究採用固態發酵，並且以原料易取得且成本低之大豆粕以及小麥粉混合，作為發酵基質，適於大量經濟化製備。發酵菌種有Aspergillus oryzae、Saccharomyces cerevisiae、Lactobacillus acidophilus、Lactobacillus casei。分為四個組別進行發酵，第一組別先以Aspergillus oryzae進行有氧發酵，假設經過酵素處理，可利於後續S. cerevisiae利用，再以S. cerevisiae進行發酵0、24、48以及72小時;第二個組別則是經過第一個組別的發酵製程後再接種乳酸菌，期降低pH值，增加保存期限；第三組別，只單獨加入酵母菌發酵0、24、48以及72小時；第四組別則將經過酵母菌發酵的樣品，再經過Lactobacillus acidophilus、Lactobacillus casei發酵24小時。發酵產品經冷凍乾燥以及萃取，進行體外細胞試驗，試驗結果顯示，有經過Aspergillus oryzae有氧發酵的組別毒性較高，因此採用低濃度60 μg/ml、30 μg/ml、15 μg/ml進行beta-casein表現試驗，結果發現經發酵的水萃液對於beta-casein的表現沒有顯著影響，第二組別經乳酸菌無氧發酵，可以降低細胞的毒性，但是對於beta-casein的表現也沒有顯著影響。直接用酵母菌發酵的水萃液，濃度125~2000 μg/ml水萃液對細胞有增生的效果，進一步進行beta-casein表現的分析，結果顯示以濃度500 μg/ml、1000 μg/ml和2000 μg/ml皆能顯著提升beta-casein表現。而酵母菌以及乳酸菌的發酵樣品，同樣在濃度500 μg/ml、1000 μg/ml和2000 μg/ml顯著提升beta-casein表現，從結果顯示當發酵時間越長則誘導beta-casein表現的量越大，且酵母菌發酵產物可以影響乳腺上皮細胞生長及beta casein的表現。

關鍵字

酵母菌 ； 乳腺上皮細胞 ； 固態發酵 ； Beta-酪蛋白

並列摘要

Mother milk plays an important role for a baby; lactation is also important for livestock. Therefore, many people want to know how to increase the milk quantity and quality. Especially, every mother wants to avoid being lack of mother milk. It was said traditional food fermented glutinous rice was good for stimulating lactation for a mother. Yeast fermented product or live yeast were also fed to animals to stimulation of lactation but scientists guess that the reasons to increase milk secretion is the yeast influenced the microflora of rumen so the digestibility become better. Our research aim to understand whether the yeast fermented products will directly influence the mammary epithelial cell and further influence the lactation. Because the solid state fermentation product is easy to dry so the research chose the solid state fermentation and used the soybean meal and wheat as material that they are cheap and easy to get. The strains used for fermentation were Aspergillus oryzae, Saccharomyces cerevisiae, Lactobacillus acidophilus and Lactobacillus casei. Aspergillus oryzae was used for aerobic fermentation; Saccharomyces cerevisiae was used for further fermentation and producing metabolic product. Finally, Lactobacillus acidophilus and Lactobacillus casei were used to lower down the pH value in order to have longer shelf time. In the research, there were four different procedures to fermentation. First, the material was aerobic fermented by A. oryzae for elevating the utilization for next step anaerobic fermentation. Then, use Saccharomyces cerevisiae anaerobic fermentation for 0 h, 24 h, 48 h and 72 h. Second, as the first procedure but add Lactobacillus acidophilus and Lactobacillus casei to anaerobic fermentation for one more day. Third, use only Saccharomyces cerevisiae to do anaerobic fermentation. Last, use Saccharomyces cerevisiae and Lactobacillus acidophilus and Lactobacillus casei do anaerobic fermentation. The results showed the Aspergillus oryzae was toxic for mammary epithelial cell. When the material was fermented only by yeast or yeast, the cell would proliferate. When treat with high concentration (500 μg/ml~2000 μg/ml) the cell will proliferation. Further analysis for beta casein expression, the results show using the extract fermented only by yeast for 48 and 72 h and concentration of 500 μg/ml to 2000 μg/ml will increase beta casein expression. The result showed the yeast fermented product could stimulate the proliferation and beta casein expression of epithelial cell and the procedure of fermentation also influenced the results. Use only aerobic fermentation by yeast or yeast and lactic acid is a suitable procedure for stimulation of lactation.

並列關鍵字

Saccharomyces cerevisiae ； mammary epithelial cell line ； solid state fermentation ； beta casein

參考文獻

李佩穎，2005，乳酸菌固態發酵黑豆產品之機能性。台北。

蘇敬哲, 食品級米麴菌發酵脫脂芝麻粕之抗氧化與 ACE 抑制活性探討. 虎尾科技大學生物科技研究所學位論文 2012, 1-46。彰化

連逸韻，2006，利用二階段固態發酵模式製備發酵豆粕以去除寡糖暨過敏性蛋白質。台中。

Smith-Kirwin, S. M.; O’Connor, D. M.; Johnston, J.; de Lancy, E.; Hassink, S. G.; Funanage, V. L., Leptin expression in human mammary epithelial cells and breast milk. The Journal of Clinical Endocrinology & Metabolism 1998, 83, 1810-1810.

Barefoot, S. F.; Klaenhammer, T. R., Detection and activity of lactacin B, a bacteriocin produced by Lactobacillus acidophilus. Appl. Environ. Microbiol. 1983, 45, 1808-1815.