本研究之目的為探討Nisin生產菌Lactococcus lactis ssp. Lactis（CCRC10791）。在發酵槽中培養時溫度、pH值控制、攪拌速率對菌體生長及Nisin生產之影響，並以自由態菌體之重複批次培養、褐藻膠鈣固定化菌體重複批次培養以及連續式培養方式操作，以期提高Nisin的生產力。
結果顯示，生產菌發酵培養生產Nisin之最佳溫度為30℃，發酵培養之pH值調控於6.5，可得到較高之Nisin活性。無論於培養時通空氣與否此乳酸菌皆會生產Nisin，通空氣培養並不會使產生之Nisin活性減少。以前一批發酵培養所得之菌體直接重複批次培養，可達到快速生產高濃度Nisin之目的。若於培養基中添加CM-Celluose或矽藻土當做Nisin之吸附劑，使發酵產生之Nisin被吸附，避免被菌體分解所放出來之蛋白質分解脢所分解，可使發酵培養後期之Nisin活性不致下降。褐藻膠鈣固定化之菌體，分別進行重複五次批次培養與連續培養，所生產之Nisin濃度仍可維持在900~1000 IU/mL之間。

Nisin production by strain Lactoccus lactis ssp. Lactis (CCRC 10791) in a 5 litter fermenter was studied. The effects of temperature, pH control and agitation speed on cell growth and nisin production were investigated. In addition, the free cells and calcium-alginate immobilized cells repeated batches were carried out to enhance the nisin productivity in the fermenter.
The optimum culture temperature for nisin production was at 30℃. Controlling the fermentation pH at pH 6.5 resulted in a higher nisin concentration. Areation would not impede the nisin production but reduced the lactate concentration. Empolying CM-cellulose or diatomaceous earth as nisin adsorbent in the culture medium could protect nisin from degradation by protease which released from lyzed cell at the end of fermentation.
The nisin produced by calcium-alginate immobilized cell during the five repeated batches fermentation and continuous culture could be maintained at concentration of 900 ~ 1000 IU/mL.

第一章 序論………………………………………………….1
1.1 研究背景簡介…………………………………….1
1.2 研究內容………………………………………….3
第二章 文獻回顧…………………………………………….6
2.1 Nisin之生化合成……………………………….…6
2.2 Nisin之抑菌特性…………………………………7
2.3 Nisin的應用……………………………..………8
2.4 Nisin之發酵生產………………………………..9
第三章 實驗材料、設備與方法…………………………….20
3.1 實驗藥品……………………………………….…..20
3.2 實驗儀器…………………………….………..……22
3.3 實驗菌株…………………………….……………..23
3.4 實驗方法與步驟…………………………………...23
第四章 結果與討論…………………………………………36
4.1 Nisin生產及分析條件探討………………………36
4.2 以發酵槽培養探討Nisin生產條件………………42
4.3 添加Nisin吸附劑對Nisin發酵生產之影響………56
4.4 以褐藻膠鈣固定化菌體生產Nisin………………61
第五章 結論與建議…………………………………………72
參考文獻…………………………………………………...74
作者簡介…………………………………………………...81

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