Enhancement of Cryoresistance and Osmotolerance of Baker's Yeast by Overexpressing Genes of Trehalose Synthase Complex

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Project title

Code/計畫編號

NSC90-2313-B019-052

Translated Name/計畫中文名

過量表現海藻糖合成脢基因以提高麵包酵母菌的抗凍性及耐滲透壓性

Project Coordinator/計畫主持人

Tsuei-Yun Fang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Food Science

Website

https://www.grb.gov.tw/search/planDetail?id=648920

Year

2001

Start date/計畫起

01-08-2001

Expected Completion/計畫迄

31-07-2002

Bugetid/研究經費

798千元

ResearchField/研究領域

食品科技(農)

本計畫利用基因重組技術，並配合酵母菌轉形技術，已獲得下列三種基因重組酵母菌: (1)過量表現TPS1 基因。(2)過量表現TPS2基因。(3)過量表現TPS1及TPS2基因。我們測量這些基因重組酵母菌與未重組酵母菌的海藻糖生成量，發現過量表現TPS1及TPS2基因者其胞內海藻糖生成較多。這些酵母菌經歷-80 ℃冷凍、72小時凍藏及30 ℃解凍的循環操作，發現三種基因重組酵母菌的存活率皆較未重組酵母菌的存活率來得高。顯示此三種基因重組酵母菌的抗凍性較未重組酵母菌為佳。在進行耐滲透壓測試時，將這些酵母菌培養於含有0、250、750、1000 mM NaCl 的YPD-agar培養基，發現在1000 mM NaCl濃度下，三種基因重組酵母菌的生長情形較未重組酵母菌稍佳。 We have created three recombinant yeast strains with different gene combinations by using the techniques of recombination DNA and yeast transformation. These three recombinant yeasts are: (1) with overexpressed TPS1 (2) with overexpressed TPS2 and (3) with both overexpressed TPS1 and TPS2. In order to examine the effects of these overexpressed genes, we determined intracellular accumulation of trehalose in wild-type and those recombinant yeasts. We found all three recombinant yeasts, which have overexpressed TPS1 and/or TPS2 genes, have produced more intracellular trehalose than wild type yeast. These yeasts were treated by freezing at -80 degree C, 72-hour frozen storage and thawing at 30 degree C. Their viabilities showed that all three recombinant yeasts survived better than wild-type yeast. This result indicates that all three recombinant yeasts have better cryoresistance than wild-type yeast. In the osmotolerance test, yeasts were grown in YPD-agar medium containing 0, 250, 750, and 1000 mM NaCl, respectively. We found three recombinant yeasts grown slightly better than wild-type yeast in the presence of 1000 mM NaCl. We have created three recombinant yeast strains with different gene combinations by using the techniques of recombination DNA and yeast transformation. These three recombinant yeasts are: (1) with overexpressed TPS1 (2) with overexpressed TPS2 and (3) with both overexpressed TPS1 and TPS2. In order to examine the effects of these overexpressed genes, we determined intracellular accumulation of trehalose in wild-type and those recombinant yeasts. We found all three recombinant yeasts, which have overexpressed TPS1 and/or TPS2 genes, have produced more intracellular trehalose than wild type yeast. These yeasts were treated by freezing at -80 degree C, 72-hour frozen storage and thawing at 30 degree C. Their viabilities showed that all three recombinant yeasts survived better than wild-type yeast. This result indicates that all three recombinant yeasts have better cryoresistance than wild-type yeast. In the osmotolerance test, yeasts were grown in YPD-agar medium containing 0, 250, 750, and 1000 mM NaCl, respectively. We found three recombinant yeasts grown slightly better than wild-type yeast in the presence of 1000 mM NaCl.

Keyword(s)

海藻糖合成脢
大量表現
麵包酵母
抗凍性
滲透壓耐受性
Trehalose synthase
Overexpression
Baker's yeast
Freezing resistance
Osmotolerance

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Enhancement of Cryoresistance and Osmotolerance of Baker's Yeast by Overexpressing Genes of Trehalose Synthase Complex

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