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首页> 外文期刊>Applied and Environmental Microbiology >Highly Efficient Production of Laccase by the Basidiomycete Pycnoporus cinnabarinus
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Highly Efficient Production of Laccase by the Basidiomycete Pycnoporus cinnabarinus

机译：拟南芥Pycnoporus cinnabarinus高效生产漆酶

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An efficient transformation and expression system was developed for the industrially relevant basidiomycete Pycnoporus cinnabarinus. This was used to transform a laccase-deficient monokaryotic strain with the homologous lac1 laccase gene placed under the regulation of its own promoter or that of the SC3 hydrophobin gene or the glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of Schizophyllum commune. SC3-driven expression resulted in a maximal laccase activity of 107 nkat ml?1 in liquid shaken cultures. This value was about 1.4 and 1.6 times higher in the cases of the GPD and lac1 promoters, respectively. lac1-driven expression strongly increased when 25 g of ethanol liter?1 was added to the medium. Accordingly, laccase activity increased to 1,223 nkat ml?1. These findings agree with the fact that ethanol induces laccase gene expression in some fungi. Remarkably, lac1 mRNA accumulation and laccase activity also strongly increased in the presence of 25 g of ethanol liter?1 when lac1 was expressed behind the SC3 or GPD promoter. In the latter case, a maximal laccase activity of 1,393 nkat ml?1 (i.e., 360 mg liter?1) was obtained. Laccase production was further increased in transformants expressing lac1 behind its own promoter or that of GPD by growth in the presence of 40 g of ethanol liter?1. In this case, maximal activities were 3,900 and 4,660 nkat ml?1, respectively, corresponding to 1 and 1.2 g of laccase per liter and thus representing the highest laccase activities reported for recombinant fungal strains. These results suggest that P. cinnabarinus may be a host of choice for the production of other proteins as well.
机译：开发了一种有效的转化和表达系统，用于工业相关的担子菌碧萝y（Pycnoporus cinnabarinus）。这被用于转化具有漆酶缺陷的单核生物菌株，该同源菌株位于其自身的启动子或SC3疏水蛋白基因或Schizophyllum commune的甘油三磷酸脱氢酶（GPD）基因的启动子的调控下。 SC3驱动的表达在液体摇动培养物中产生最大的漆酶活性为107 nkat ml？1。在GPD和lac1启动子的情况下，该值分别约为1.4和1.6倍。当将25 g乙醇升？1添加到培养基中时，lac1驱动的表达会大大增加。因此，漆酶活性增加到1,223nkat·ml-1。这些发现与乙醇诱导某些真菌中漆酶基因表达的事实相吻合。值得注意的是，当在SC3或GPD启动子后表达lac1时，在25 g乙醇升？1的存在下，lac1 mRNA的积累和漆酶活性也大大增加。在后一种情况下，获得最大的漆酶活性为1393纳卡·毫升·l（即360毫克/升·l）。在40 g乙醇升？1的存在下生长，在其自身启动子或GPD后面表达lac1的转化子中，漆酶的产量进一步增加。在这种情况下，最大活性分别为3,900和4,660 nkat ml？1，对应于每升1和1.2克漆酶，因此代表了重组真菌菌株报道的最高漆酶活性。这些结果表明，朱砂疟原虫可能也是产生其他蛋白质的众多选择。

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