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首页> 外文期刊>Applied Microbiology and Biotechnology >High-level expression of aryl-alcohol oxidase 2 from Pleurotus eryngii in Pichia pastoris for production of fragrances and bioactive precursors
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High-level expression of aryl-alcohol oxidase 2 from Pleurotus eryngii in Pichia pastoris for production of fragrances and bioactive precursors

机译：来自PICHIA Pastoris的Pleurotus的芳基醇氧化酶2的高水平表达，用于生产香料和生物活性前体

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摘要

The fungal secretome comprises various oxidative enzymes participating in the degradation of lignocellulosic biomass as a central step in carbon recycling. Among the secreted enzymes, aryl-alcohol oxidases (AAOs) are of interest for biotechnological applications including production of bio-based precursors for plastics, bioactive compounds, and flavors and fragrances. Aryl-alcohol oxidase 2 (PeAAO2) from the fungusPleurotus eryngiiwas heterologously expressed and secreted at one of the highest yields reported so far of 315 mg/l using the methylotrophic yeastPichia pastoris(recently reclassified asKomagataella phaffii). The glycosylated PeAAO2 exhibited a high stability in a broad pH range between pH 3.0 and 9.0 and high thermal stability up to 55 degrees C. Substrate screening with 41 compounds revealed that PeAAO2 oxidized typical AAO substrates likep-anisyl alcohol, veratryl alcohol, andtrans,trans-2,4-hexadienol with up to 8-fold higher activity than benzyl alcohol. Several compounds not yet reported as substrates for AAOs were oxidized by PeAAO2 as well. Among them, cumic alcohol and piperonyl alcohol were oxidized to cuminaldehyde and piperonal with high catalytic efficiencies of 84.1 and 600.2 mM(-1) s(-1), respectively. While the fragrance and flavor compound piperonal also serves as starting material for agrochemical and pharmaceutical building blocks, various positive health effects have been attributed to cuminaldehyde including anticancer, antidiabetic, and neuroprotective effects. PeAAO2 is thus a promising biocatalyst for biotechnological applications.
机译：真菌沉淀包括参与木质纤维素生物质的降解作为碳回收的中央步骤的各种氧化酶。在分泌的酶中，芳基 - 醇氧化酶（AAOS）对生物技术应用感兴趣，包括生产塑料，生物活性化合物和香料和香料的生物基前体。来自FunGUSPUROTUS的芳基 - 醇氧化酶2（PHAOR2）异常表达并在迄今为止迄今为止的最高产量中分泌315 mg / L的含量，使用甲基营养酵母酵母（最近重新分类Askomagatabla phaffii）。糖基化的PeaO 2在pH 3.0和9.0之间的宽pH范围内表现出高稳定性，并且高达55℃的热稳定性高达55℃。用41种化合物的底物筛选显示Peao2氧化典型的AaO底物LiblP-吲哚醇，戊醇，醇，反式-2,4-十六二烯醇，活性高达8倍，比苄醇更高。尚未报道的几种化合物作为AAOS的底物被PeaO 2氧化。其中，分别具有84.1和600.2mm（-1）（-1）的高催化效率，累积醇和哌隆醇和哌啶醇和哌啶，分别具有84.1和600.2mm（-1）的高催化效率。虽然香料和风味复合哌啶也用作农用化学和药物积木的原料，而各种阳性健康效应归因于抗癌醛，包括抗癌，抗糖尿病和神经保护作用。因此，PeaO2是生物技术应用的有前途的生物催化剂。

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