Effect of oxygen supply on passaging, stabilising and screening of recombinant Hansenula polymorpha production strains in test tube cultures
Effect of oxygen supply on passaging, stabilising and screening of recombinant Hansenula polymorpha production strains in test tube cultures
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DOI：
10.1016/s1567-1356(03)00147-8
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发表时间：
2003-11-01
影响因子：
3.2
通讯作者：
Büchs, J
中科院分区：
文献类型：
--
作者：
Stöckmann, C;Losen, M;Büchs, J
将24个多形汉逊酵母转化体在葡萄糖培养基中传代和稳定，并在摇动试管中在甘油培养基中筛选重组植酸酶。在有限或非有限的氧气供应下进行培养。用亚硫酸盐氧化法测定了试管的最大氧传递能力。限氧葡萄糖培养物导致部分厌氧代谢并形成4.1 g乙醇l（-1），随后进行需氧代谢。无限制的供氧导致溢流代谢和积累2.1 g乙酸l（-1），降低生物量产量。在筛选主培养物中使用甘油可防止副产物形成，而与氧气供应无关。在葡萄糖培养基中在非限制性氧气供应下预培养导致筛选主培养物的20小时滞后期。当在氧限制下进行预培养时，未观察到该滞后期。植酸酶活性平均高出25%，在有限的氧气供应下传代，稳定和筛选的培养物比在非有限的氧气供应下的培养物。(C)2003年，欧洲微生物学会联合会。Elsevier B. V.出版，保留所有权利。
Twenty-four Hansenula polymorpha transformants were passaged and stabilised in glucose medium and screened in glycerol medium for recombinant phytase in shaken test tubes. The cultivations were performed under either limited or non-limited oxygen supply. Maximum oxygen transfer capacities of test tubes were assessed by sulfite oxidation. Oxygen-limited glucose cultures resulted in a partially anaerobic metabolism and formation of 4.1 g ethanol l(-1), which was subsequently aerobically metabolised. Non-limited oxygen supply led to overflow metabolism and to accumulation of 2.1 g acetic acid l(-1), reducing the biomass yield. The use of glycerol in the screening main cultures prevented by-product formation irrespective of oxygen supply. Preculturing in glucose medium under non-limited oxygen supply resulted in a 20-h lag phase of the screening main culture. This lag phase was not observed when preculturing was performed under oxygen limitation. Phytase activity was on average 25% higher in cultures passaged, stabilised and screened under limited oxygen supply than in cultures under non-limited oxygen supply. (C) 2003 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.