04 1 bioreactor Bioprocess Engineering PCR host selection
04. 1 養菌&bioreactor Bioprocess Engineering 基因重組� PCR, host selection� 或微生物篩選� plasmid selection� 前處理� 殺菌等� 生化反應器 Lab. 生產 Recombinant protein metabolites 細胞固定化� 酵素固定化� 廠生產 破碎細胞 分離及純化
medium(培養基) rich media defined media M 9 medium: Na 2 HPO 4‧ 7 H 2 O 磷源 KH 2 PO 4 磷源 Na. Cl NH 4 Cl 氮源 Mg. SO 4 硫源 Ca. Cl 2 glucose 碳源 有時Na、K、Mg、Ca等金屬離子可作為Coemzyme (輔�)其所需量 很少。 LB medium: Tryptone-b Yeast extract-B Na. Cl
colony
Determining cell mass dry weight optical density (O. D. ) spectrophotometer O. D. 550 O. D. 600
圖表. 1 Growth curve of cell Time (hr)
圖表. 2 Growth curve of cell Time (hr)
計算: Maximum growth rate ……………. (1) when is constant 由plot log OD vs. t The greatest slope in linear region: 由實驗數據得知: ……. (2)
計算: =doubling time of cells ……………. (3) occurs when ……………. (4)
� 二� 生化反應器 1. Ferment or 發酵槽 � batch� 2. chemosta t
Chemostat: cellular growth is usually limited by one essential nutrient, and other nutrients are in excess (P 183, 184)
Turbidostat: The cell concentration in the culture vessel is maintained constant by monitoring the optical density of the culture and controlling the feed flow pate. (P 184)
The ideal chemostat:
使用chemostat為bioreactor,其產量將遠 大於batch。可是為什麼 業界使用batch的比 率遠高於chemostat? 1. Secondary products 2. Genetic instability 3. Operability and reliability 4. Market economics
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