Heterologous production of a laccase from the basidiomycete Pycnoporus cinnabarinus in the dimorphic yeast Yarrowia lipolytica
Heterologous production of a laccase from the basidiomycete Pycnoporus cinnabarinus in the dimorphic yeast Yarrowia lipolytica
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DOI：
10.1016/j.femsyr.2004.10.009
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发表时间：
2005-04-01
影响因子：
3.2
通讯作者：
Beckerich, JM
中科院分区：
文献类型：
--
作者：
Madzak, C;Otterbein, L;Beckerich, JM
朱砂密孔菌lac1基因在解脂耶氏酵母中表达。测试了不同的分泌信号和培养基。产量与培养物生长速率和细胞形态（在低生长速率下最高，没有菌丝体）两者相关。对重组漆酶进行表征（免疫检测、N末端测序）并纯化。在生物反应器中，产量估计为20 mg l（-1）。因此，复杂的金属酶可以在耶氏酵母中产生，假设宿主生理学的一些控制。Lac1p生产在Yarrowia，毕赤酵母和曲霉属进行了比较：重组蛋白是有活性的，但宿主系统在转化效率，生产和糖基化方面不同。如果不是最好的生产者，Yarrowia提供了非常高的转化效率，允许将漆酶的基因工程用于工业应用。(c)2004年，欧洲微生物学会联合会。Elsevier B.V.出版，保留所有权利。
Pycnoporus cinnabarinus lac1 gene was expressed in Yarrowia lipolytica. Different secretion signals and culture media were tested. Production was correlated to both culture growth rate and cell morphology (highest at low growth rate, without mycelium). Recombinant laccase was characterized (immunodetection, N-terminal sequencing) and purified. Production was estimated to 20 mg l(-1) in a bioreactor. Thus, complex metalloenzymes can be produced in Yarrowia, assuming some control of host physiology. Lac1p production was compared in Yarrowia, Pichia and Aspergillus: recombinant proteins were active, but host systems differed in transformation efficiency, production, and glycosylation. If not the best producer, Yarrowia offers very high transformation efficiencies, allowing the genetic engineering of laccases for industrial applications. (c) 2004 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.