黑曲霉N402和葡萄糖淀酵酶高产转化体在恒化和循环培养物中生长和产物形成,提供了多个拷贝的glaA基因。

J M Schrickx, A S Krave, J C Verdoes, C A van den Hondel, A H Stouthamer, H W van Verseveld
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引用次数: 74

摘要

对黑曲霉N402野生型和葡萄糖淀粉酶高产型进行连续和循环培养,研究其生长和产物形成特性。在摇瓶培养中,转化产生的葡萄糖淀粉酶的数量大约是野生型菌株的五倍。与这些结果相反,在葡萄糖限制的连续培养中发现了两倍的过量生产,而在麦芽糊精限制的连续培养中没有发现过量生产。可以区分出两个特定生长速率区域,一个特定生长速率较低(区域I),一个特定生长速率高于0.12 h-1(区域II)。在区域I中,观察到菌丝形态和分生孢子形成的变化。由此得出结论,维护需求取决于整个测量生长速率范围内的特定生长速率。在对丝状真菌进行连续培养时,应考虑由这种现象引起的基质利用线性方程的线性偏差。在循环培养中,木糖作为限制碳源对葡萄糖淀粉酶的产生有很强的抑制作用。在麦芽糖糊精限制下发现了5倍的过量生产。约150 h后,葡萄糖淀粉酶的总生成量仍在增加,而以碳为单位的总生成量保持不变。在此之后,未观察到生物量的增加。这些结果表明,自溶和隐生长发生在循环发酵中,细胞死亡率等于生长速度。
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Growth and product formation in chemostat and recycling cultures by Aspergillus niger N402 and a glucoamylase overproducing transformant, provided with multiple copies of the glaA gene.

Continuous and recycling cultures were carried out with Aspergillus niger N402 wild-type and a glucoamylase overproducing transformant to investigate growth and product formation characteristics. In shake flask cultures, the amount of glucoamylase produced by the transformant was about five times more than by the wild-type strain. In contrast with these results, a twofold overproduction was found in glucose-limited continuous cultures, while no overproduction was found under maltodextrin-limitation. Two regions of specific growth rates could be distinguished, one at specific growth rates lower (domain I) and one at specific growth rates higher than 0.12 h-1 (domain II). In domain I changes in mycelium morphology and conidia formation were observed. It has been concluded that maintenance requirements are dependent on the specific growth rate over the whole range of measured growth rates. The deviation in linearity in the linear equation of substrate utilization, caused by this phenomenon, should be considered when continuous cultures with filamentous fungi are performed. In recycling cultures, xylose as limiting carbon source repressed glucoamylase production very strongly. Under maltodextrin-limitation a fivefold overproduction was found. After about 150 h , the total amount of glucoamylase produced was still increasing, while total amount of product, measured as carbon, remained constant. After this time no increase in the amount of biomass formed was observed. These results suggest autolysis and cryptic growth taking place in a recycling fermenter and cell death rate equalling growth rate.

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