利用紫外光誘發Leuconostocmesenteroides突變以提高酒精發酵能力

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Please use this identifier to cite or link to this item: http://140.128.103.80:8080/handle/310901/4211

Title:	利用紫外光誘發Leuconostocmesenteroides突變以提高酒精發酵能力
Other Titles:	Improvement of the ethanol productivity of Leuconostoc mesenteroides by screening high productivity strains from U.V. light induced mutants
Authors:	詹佳育 Chan, Chia-Yu
Contributors:	李根永 Li, Ken-Yuon 東海大學食品科學系
Keywords:	紫外光突變;人工變異;反應曲面法;最適化;Leuconostoc mesenteroides;乳酸菌 mutation;RSM;Leuconostoc mesenteroides;lactic acid bacteria;U.V.
Date:	2002
Issue Date:	2011-05-19T05:47:13Z (UTC)
Abstract:	本研究採用分離自甘蔗汁之異質發酵型乳酸菌Leuconostoc mesenteroides，發酵甘蔗汁製造低酒精乳酸飲料。Ln. mesenteroides可產生酒精，但酒精產量偏低，故在本實驗中先以紫外光誘導Ln. mesenteroides突變，再從變異株中篩選酒精產量較高的菌株。經過六次突變，以隨機及酒精耐性株中篩選，從506支突變株中篩出一支酒精產量高於母株11 %之突變株，命名為Ln. mesenteroides THF。在培養基研究方面，首先，以Plackett- Burman實驗設計探討甘蔗汁、半胱胺酸、小麥生胚芽、啤酒酵母、天數、大豆蛋白腖(Soybean peptone)、葡萄糖等七個因子，對酒精發酵影響。結果得知關鍵因子有甘蔗汁、半胱胺酸、小麥生胚芽及蛋白?。之後以此四因子進行部份因子實驗設計以求得各因子之主效應，再以陡升路徑實驗快速求得極值所在區域，然後，以中心混成實驗設計所得之數據，利用SAS軟體進行分析後，得到二次方程式： Y＝-764.3769＋2.3265X1＋136.2780X2＋414.5118X3－0.0773X1X1＋0.3395X2X1－13.4898X2X2＋0.8470X3X1＋5.3100X3X2－116.6578X3X3 將此二次方程式轉換得正則方程式為： Y = 112.8－4.943Z12－9.822Z22－20.98Z32 此時Y為酒精生產力，Z1為甘蔗汁，Z2為小麥生胚芽，而Z3則為peptone。由迴歸方程式求得極大值，各成分對應濃度為：甘蔗汁~39.38 %(v/v)，小麥生胚芽~5.95 %(w/v)，蛋白腖~2.06 %(w/v)。在最適培養基Ln. mesenteroides THF經兩天發酵酒精產量可達110.96mM (0.51%)，與預測值無明顯差異。 The objective of this research is to improve the ethanol productivity of Leuconostoc mesenteroides in cane juice fermentation. A strain of Ln. mesenteroides isolated originally from cane juice was induced to mutate by irradiating with U.V. light. Having screened 506 mutants, a mutant with ethanol productivity of 11 % higher than the parent strain was selected. The mutant was named Ln. mesenteroides THF. To customized a optimum medium for increasing the ethanol production of Ln. mesenteroides THF, a series of medium study was carried out. Firstly, an experiment of the Plackett- Burman design was run to decide the major ingredients for ethanol fermention. The result showed that cane juice, wheat germ, cysteine, and soybean peptone were significant factors. Secondly, an experiment of 24-1 fractional fractiorial design was performed to find the path approaching to the optimum concentrations of cane juice, wheat germ, and peptone. Following the result of fractional fractiorial experiment, an experiment based on the central composite design was fulfilled and a regression quadratic equation was developed. Y＝-764.3769＋2.3265X1＋136.2780X2＋414.5118X3－0.0773X1X1＋0.3395X2X1－13.4898X2X2＋0.8470X3X1＋5.3100X3X2－116.6578X3X3 Then, the equation was transformed to a canonical equation： Y = 112.8－4.943Z12－9.822Z22－20.98Z32 According to the regression equation, the optimum concentrations of the medium ingredients, cane juice, Wheat germ, and peptone were determined as 39.38 %, 5.95 %, and 2.06 %, respectively.
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