Dynamic flux balance analysis for pharmaceutical protein production by Pichia pastoris: Human growth hormone

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• 作者：Pı ; nar Ç ; alı ; k ; ^a ; ^{pcalik@metu.edu.tr} ; Merve &#x15e ; ahin^a ; Hatice Ta&#x15f ; pı ; nar¹ ; ^a ; Elif &#x15e ; . Soyaslan¹ ; ^a ; Bahar İ ; nankur¹ ; ^a
• 关键词：Pichia pastoris ; Flux ; Sorbitol ; Methanol ; Metabolism ; Fed-batch ; Human growth hormone
• 刊名：Enzyme and Microbial Technology
• 出版年：2011
• 出版时间：7 March 2011
• 年：2011
• 卷：48
• 期：3
• 页码：209-216
• 全文大小：433 K

文摘

The influence of methanol feeding rate on intracellular reaction network of recombinant human growth hormone (rhGH) producing Pichia pastoris was investigated at three different specific growth rates, namely, 0.02 (MS-0.02), 0.03 (MS-0.03), and 0.04 h⁻¹ (MS-0.04) where Period-I (33 ≤ t < 42 h) includes the early exponential growth phase; Period-II (42 ≤ t < 48 h) is the exponential growth phase where the specific cell growth rate decreases; Period-III (48 ≤ t ≤ 51 h) is the exponential growth phase where rhGH concentration was the highest; and Period-IV (t > 51 h) is the diminution phase for rhGH and cell synthesis. In Period-I, almost all of the formaldehyde entered the assimilatory pathway, at MS-0.02 and MS-0.03, whereas, at MS-0.04 high methanol feeding rate resulted in an adaptation problem. In Period-III, only at MS-0.02 co-carbon source sorbitol uptake-flux was active showing that sorbitol uptake does not affected from the predetermined feeding rate of methanol at μ₀ > 0.02 h⁻¹. The biomass synthesis flux value was the highest in Period-I, -II and -III, respectively at MS-0.03 & MS-0.04, MS-0.04 and MS-0.02; whereas, rhGH flux was the highest in Period-I, -II, and -III, respectively at MS-0.03, MS-0.02 and MS-0.03. Based on the fluxes, Period-I should start with MS-0.03 methanol feeding rate and starting from the middle of Period-II methanol feeding rate should be shifted to MS-0.02.