Koda Akio, Bogaki Takayuki, Minetoki Toshitaka, Hirotsune Masato
General Research Laboratory, Ozeki Co., Ltd., 4-9 Imazu, Nishinomiya-shi, Hyogo 663-8227, Japan.
J Biosci Bioeng. 2005 Nov;100(5):531-7. doi: 10.1263/jbb.100.531.
We describe the successful heterologous expression of the Solanum tuberosum alpha-glucan phosphorylase (GP) gene in Aspergillus niger. Special attention was paid to the influence of different codon usage and A+T content in the coding region on GP protein expression. Use of A. niger-preferred codon usage and lower A+T content in a synthetic gene (GP-syn) resulted in a significant improvement in the level of the GP mRNA and a dramatic increase in the quantity of GP protein produced such that it accounted for approximately 10% of the total soluble protein. We suggest that redesigning the primary DNA sequence encoding a desired protein product can be an extremely effective method for improving heterologous protein production in filamentous fungi.
我们描述了马铃薯α-葡聚糖磷酸化酶(GP)基因在黑曲霉中的成功异源表达。特别关注了编码区不同密码子使用情况和A+T含量对GP蛋白表达的影响。在合成基因(GP-syn)中使用黑曲霉偏好的密码子使用方式和较低的A+T含量,导致GP mRNA水平显著提高,所产生的GP蛋白数量急剧增加,使其占总可溶性蛋白的约10%。我们认为,重新设计编码所需蛋白质产物的初级DNA序列可能是提高丝状真菌中异源蛋白质产量的一种极其有效的方法。