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Journal of Industrial Microbiology & Biotechnology, 2002, 29 : 185~188 指導老師：李重義 報告學生：王嗣復

Isolation of a mutant strain of Pseudomonas sp. ATCC 31461 exhibiting elevates polysaccharide production. Journal of Industrial Microbiology & Biotechnology, 2002, 29 : 185~188 指導老師：李重義 報告學生：王嗣復. Introduction. Gellan 由 Pseudomonas 產生的一種胞外多 醣

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Journal of Industrial Microbiology & Biotechnology, 2002, 29 : 185~188 指導老師：李重義 報告學生：王嗣復

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  1. Isolation of a mutant strain of Pseudomonas sp. ATCC 31461 exhibiting elevates polysaccharide production Journal of Industrial Microbiology & Biotechnology, 2002, 29:185~188 指導老師：李重義 報告學生：王嗣復

  2. Introduction • Gellan由Pseudomonas產生的一種胞外多醣 • 單元為20% glucuronic acid, 60% glucose,20% rhamnose

  3. Pseudomonas ATCC 31461 具有相當高的產量 • 利用Glucose, corn syrup等作為碳源可提高產量 • 由具rifampicin抗性之Strain R40分離出Strain MP1, 產量較高 • 目的利用突變劑及抗生素篩選出產量較高的突變株

  4. 材料方法 • 菌株：Strain ATCC 31461 • 培養基：以Glucose或corn syrup作為唯一碳源之無機鹽培養基 • 突變劑：ethyl methyl sulfonate (EMS) • 抗生素：ampicillin

  5. 菌株培養於NB中, 生長至對數期時加入1% EMS 靜置培養60 min • 以1:30之比例使用NB稀釋後, 250 rpm隔夜培養存活率約為2.2% • 取107之突變株spread到含有100 ppm Ampicillin之NA上培養72 hrs • 觀察菌落產生多醣的情況, 自2250個colony中選出23個接種至含有3% Glucose之無機鹽培養機中培養48 hrs • 最後篩選出一株產量最佳者, 命名為EGP-1

  6. 篩選方式 • 為有目標的突變篩選 • 突變劑處理後, 對抗生素具有抗性者：推測對於Ampicillin具有抗性者, 胞外多醣的分泌應較佳, 可阻擋Ampicillin攻擊細胞壁 • 以Glucose作為唯一碳源可增進胞外多醣的表現可作為結果篩選的依據

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