el-Enshasy H, Hellmuth K, Rinas U
GBF National Research Center for Biotechnology, Biochemical Engineering Division, Braunschweig, Germany.
Appl Biochem Biotechnol. 1999 Jul;81(1):1-11. doi: 10.1385/abab:81:1:1.
The effect of culture conditions such as medium composition and shear stress on the fungal pellet morphology in shake-flask cultures and its relation to glucose oxidase (GOD) excretion by recombinant Aspergillus niger NRRL 3 (GOD 3-18) was investigated. It was shown that culture conditions resulting in the formation of smaller fungal pellets with an increased mycelial density result in higher yields of exocellular GOD. The pellets obtained in shake-flask cultures showed distinct layers of mycelial density with only the thin outer layer consisting of a dense mycelial network. The performance of the recombinant strain and the process of pellet formation was also analyzed during batch cultivation in a stirred-tank bioreactor. It was shown that the process of pellet formation occurred in two steps: (1) aggregation of free spores to spore clusters with subsequent germination and formation of small aggregates surrounded by a loose hyphal network, and (2) aggregation of the primary aggregates to the final full-size pellets. The fungal pellets formed during bioreactor cultivation were smaller, did not show large differences in mycelial density, and were more efficient with respect to the production of exocellular GOD. The decreasing pellet size also correlated with an increased mycelial density, indicating an improvement of the transport of nutrients to the inner parts of the pellet.
研究了培养基成分和剪切应力等培养条件对摇瓶培养中真菌菌球形态的影响及其与重组黑曲霉NRRL 3(GOD 3-18)分泌葡萄糖氧化酶(GOD)的关系。结果表明,培养条件导致形成菌丝体密度增加的较小真菌菌球,从而使胞外GOD产量更高。摇瓶培养中获得的菌球显示出明显的菌丝体密度层,只有薄薄的外层由密集的菌丝体网络组成。在搅拌罐生物反应器的分批培养过程中,还分析了重组菌株的性能和菌球形成过程。结果表明,菌球形成过程分两步进行:(1)游离孢子聚集成孢子簇,随后萌发并形成被松散菌丝网络包围的小聚集体;(2)初级聚集体聚集成最终的全尺寸菌球。生物反应器培养过程中形成的真菌菌球较小,菌丝体密度差异不大,并且在胞外GOD生产方面更高效。菌球尺寸的减小也与菌丝体密度的增加相关,表明营养物质向菌球内部的运输得到改善。