The Manipulation of Trehalose Synthesis and Hydrolysis Related Genes to Enhance the Cryoresistance and Osmotolerance of Baker´s Yeast

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Project title

Code/計畫編號

NSC91-2313-B019-038

Translated Name/計畫中文名

操控海藻糖合成及分解相關基因以改進麵包酵母菌之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=757838

Year

2002

Start date/計畫起

01-08-2002

Expected Completion/計畫迄

01-07-2003

Bugetid/研究經費

795千元

ResearchField/研究領域

生物技術(農)
食品科技(農)

本計畫利用前二個計畫內所獲得之基因刪除重組酵母菌及含TPS1或TPS2基因表現質體，以酵母菌轉形技術，將TPS1或TPS2基因大量表現在NTH1基因已刪除之重組酵母菌、ATH1基因已刪除之重組酵母菌、或NTH1及ATH1基因均已刪除之重組酵母菌中。這些不同的基因組合，包括: (1)過量表現TPS1基因，並且刪除NTH1基因。(2)過量表現TPS1基因，並且刪除ATH1基因。(3)過量表現TPS1基因，並且同時刪除NTH1及ATH1基因。(4)過量表現TPS2基因，並且刪除NTH1基因。(4)過量表現TPS2基因，並且刪除 ATH1基因。(5)過量表現TPS2基因，並且同時刪除NTH1 及ATH1基因。(6)同時過量表現TPS1及TPS2基因，並刪除NTH1基因。(7)同時過量表現TPS1及TPS2基因，並且刪除ATH1基因。(8)同時過量表現TPS1及TPS2基因，並且同時刪除NTH1及ATH1基因。我們測量這些具有不同基因組合之重組酵母菌與控制組野生型酵母菌的海藻糖生成量發現，大部份的重組酵母菌其胞內海藻糖生成量較控制組野生型酵母菌多。這些酵母菌經歷-80 degree C冷凍、24小時凍藏及30 degree C解凍的循環操作，發現重組酵母菌的存活率與其海藻糖生成量無關。在進行耐滲透壓測試時，將這些酵母菌培養於含有0-1000 mM Sucrose的YPD-agar培養基，發現在基因重組酵母菌的生長情形與控制組野生型酵母菌相似。 We have created nine recombinant yeast strains with different gene combinations by using the techniques of recombination DNA and yeast transformation. These nine recombinant yeasts are: (1) with overexpressed TPS1 and deleted NTH1 (2) with overexpressed TPS1 and deleted ATH1 (3) with overexpressed TPS1 and deleted both NTH1 and ATH1 (4) with overexpressed TPS2 and deleted NTH1 (5) with overexpressed TPS2 and deleted ATH1 (6) with overexpressed TPS1, and deleted both NTH1 and ATH1 (7) with overexpressed both TPS1 and TPS2, and deleted NTH1 (8) with overexpressed both TPS1 and TPS2, and deleted ATH1 (9) with overexpressed both TPS1 and TPS2, and deleted both NTH1 and ATH1. In order to examine the effects of these overexpressed and deleted genes, we determined intracellular accumulation of trehalose in wild-type and those recombinant yeasts. We found most recombinant yeasts, which have overexpressed or deleted genes, have produced more intracellular trehalose than wild type yeast. These yeasts were treated by freezing at -80 degree C, 24-hour frozen storage and thawing at 30 degree C. Their viabilities showed that the viabilities were not related to the trehalose contents. In the osmotolerance test, yeasts were grown in YPD-agar medium containing 0, 250, 750 and 1000 mM sucrose, respectively. We found recombinant yeasts grown similar to wild-type yeast in all test conditions.

Keyword(s)

Trehalose
Baker' s yeast
Trehalose-6-phosphate synthase
Trehalose-6-phosphate phosphatase
Acid trehalase
Neutral trehalase
Cryoresistance
Osmotolerance

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The Manipulation of Trehalose Synthesis and Hydrolysis Related Genes to Enhance the Cryoresistance and Osmotolerance of Baker´s Yeast

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