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漆酶是一种用于酵母中合成基因电路的多功能蛋白质。

Laccase is a multitasking protein for synthetic gene circuits in the yeast .

作者信息

Yu Lifang, Asemoloye Michael Dare, Marchisio Mario Andrea

机构信息

School of Pharmaceutical Science and Technology, Tianjin University, 300072, Tianjin, China.

出版信息

Synth Syst Biotechnol. 2024 May 14;9(4):638-646. doi: 10.1016/j.synbio.2024.05.007. eCollection 2024 Dec.

Abstract

Laccase is a multicopper oxidase enzyme that oxidizes a variety of substrates, including polyphenols and polycyclic aromatic hydrocarbons (PAHs). It catalyzes the four-electron reduction of molecular oxygen that results in the production of water as a by-product. Thus, laccase can play an important role in environmental care. Previously, we have successfully expressed laccase (TtLcc1) in the yeast . In this work, we have expressed in yeast another laccase, LacA from sp. AH28-2, and tested its function on PAHs. Yeast cells engineered to produce the two laccases performed efficient PAH degradation. Both TtLcc1 and LacA led to the construction of spatiotemporal fluorescence-pulse generators when combined with a benzoate/salicylate yeast biosensor in a two-population system. Moreover, laccases returned a visual output signal in yeast synthetic circuits-upon reacting with ABTS (2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid)). Thus, in , laccases are a powerful alternative to fluorescent reporter proteins.

摘要

漆酶是一种多铜氧化酶,可氧化多种底物,包括多酚和多环芳烃(PAHs)。它催化分子氧的四电子还原反应,副产物是水。因此,漆酶在环境保护中可发挥重要作用。此前,我们已在酵母中成功表达了漆酶(TtLcc1)。在这项工作中,我们在酵母中表达了另一种来自AH28 - 2菌株的漆酶LacA,并测试了其对多环芳烃的功能。经基因工程改造以产生这两种漆酶的酵母细胞能高效降解多环芳烃。在双群体系统中,当TtLcc1和LacA与苯甲酸/水杨酸酵母生物传感器结合时,二者都能构建时空荧光脉冲发生器。此外,漆酶与ABTS(2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸))反应后,能在酵母合成电路中产生视觉输出信号。因此,在[具体语境未明确]中,漆酶是荧光报告蛋白的有力替代品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7762/11112005/0bf614b618af/gr1.jpg

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