利用糙皮侧耳（Agrocybe aegerita）生产非特异性过氧化物酶，对生物乙醇副产物进行增值利用：技术和蛋白质组学视角。

Valorization of bioethanol by-products to produce unspecific peroxygenase with Agrocybe aegerita: Technological and proteomic perspectives.

机构信息

CRETUS, Department of Chemical Engineering, Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Galicia, Spain.

出版信息

N Biotechnol. 2023 Sep 25;76:63-71. doi: 10.1016/j.nbt.2023.05.001. Epub 2023 May 9.

DOI:10.1016/j.nbt.2023.05.001

PMID:37169331

Abstract

Unspecific peroxygenase (UPO) presents a wide range of biotechnological applications. This study targets the use of by-products from bioethanol synthesis to produce UPO by Agrocybe aegerita. Solid-state and submerged fermentations (SSF and SmF) were evaluated, achieving the highest titers of UPO and laccase in SmF using vinasse as nutrients source. Optimized UPO production of 331 U/L was achieved in 50% (v:v) vinasse with an inoculum grown for 14 days. These conditions were scaled-up to a 4 L reactor, achieving a UPO activity of 265 U/L. Fungal proteome expression was analyzed before and after UPO activity appeared by shotgun mass spectrometry proteomics. Laccase, dye-decolorizing peroxidases (DyP), lectins and proteins involved in reactive oxygen species (ROS) production and control were detected (in addition to UPO). Interestingly, the metabolism of complex sugars and nitrogen sources had a different activity at the beginning and end of the submerged fermentation.

摘要

非特异性过氧化物酶 (UPO) 具有广泛的生物技术应用。本研究旨在利用生物乙醇合成的副产物，通过 Agerita 银蕈生产 UPO。评估了固态发酵和浸没发酵 (SSF 和 SmF)，使用酒糟作为营养源，在 SmF 中实现了 UPO 和漆酶的最高效价。在 50%（v:v）酒糟中，用培养 14 天的接种物实现了 331 U/L 的优化 UPO 产量。这些条件被放大到 4 L 反应器中，实现了 265 U/L 的 UPO 活性。通过鸟枪法质谱蛋白质组学分析了 UPO 活性出现前后真菌蛋白质组的表达。检测到漆酶、染料脱色过氧化物酶 (DyP)、凝集素以及与活性氧 (ROS) 产生和控制相关的蛋白质（除 UPO 外）。有趣的是，在浸没发酵的开始和结束时，复杂糖和氮源的代谢具有不同的活性。