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在仅供应空气的氧气控制发酵罐中,嗜铁钩端螺旋菌MSR-1实现高产生长和磁小体形成。

High-yield growth and magnetosome formation by Magnetospirillum gryphiswaldense MSR-1 in an oxygen-controlled fermentor supplied solely with air.

作者信息

Sun Jian-Bo, Zhao Feng, Tang Tao, Jiang Wei, Tian Jie-sheng, Li Ying, Li Ji-Lun

机构信息

State Key Laboratories for Agrobiotechnology and College of Biological Sciences, China Agricultural University, Beijing, People's Republic of China.

出版信息

Appl Microbiol Biotechnol. 2008 Jun;79(3):389-97. doi: 10.1007/s00253-008-1453-y. Epub 2008 Apr 19.

Abstract

Magnetotactic bacteria are difficult to grow under defined conditions in culture, which has presented a major obstacle to commercial application of magnetosomes. We studied the relationships among the cell growth, magnetosome formation, dissolved oxygen concentration (DO), and the ability to supply oxygen to the cells. Mass culture of Magnetospirillum gryphiswaldense MSR-1 for the production of magnetosomes was established in a 42-L fermentor under the following conditions: (1) sterile air was the sole gas supplied in the fermentor, and DO could be regulated at any level below 10% saturation by cascading the stir rate to DO, (2) to resolve the paradoxical situation that the cell growth requires higher DO whereas magnetosome formation requires low DO below the detectable range of regular oxygen electrode, DO was controlled to optimal level using the change of cell growth rate, rather than reading from the highly sensitive oxygen electrode, as the signal for determining appropriate DO, and (3) timing and rate of supplying the substrates were determined by measuring cell density and Na-lactate concentration. Under these conditions, cell density (OD565) of strain MSR-1 reached 7.24 after 60-h culture in a 42-L fermentor, and cell yield (dry weight) was 2.17 g/L, the highest yield so far being reported. The yield of magnetosomes (dry weight) was 41.7 mg/L and 16.7 mg/L/day, which were 2.8 and 2.7 times higher than the previously reported yields.

摘要

趋磁细菌在特定培养条件下难以生长,这给磁小体的商业应用带来了重大障碍。我们研究了细胞生长、磁小体形成、溶解氧浓度(DO)以及向细胞供氧能力之间的关系。在42升发酵罐中,在以下条件下建立了嗜格孢磁螺菌MSR-1大规模培养以生产磁小体:(1)无菌空气是发酵罐中供应的唯一气体,通过将搅拌速率与溶解氧级联,可将溶解氧调节至10%饱和度以下的任何水平;(2)为解决细胞生长需要较高溶解氧而磁小体形成需要低于常规氧电极可检测范围的低溶解氧这一矛盾情况,以细胞生长速率的变化而非从高灵敏度氧电极读取数据作为确定合适溶解氧的信号,将溶解氧控制在最佳水平;(3)通过测量细胞密度和乳酸钠浓度来确定底物供应的时间和速率。在这些条件下,菌株MSR-1在42升发酵罐中培养60小时后,细胞密度(OD565)达到7.24,细胞产量(干重)为2.17克/升,这是目前报道的最高产量。磁小体产量(干重)为41.7毫克/升,日产量为16.7毫克/升,分别比先前报道的产量高2.8倍和2.7倍。

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