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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.

DOI:10.1016/j.synbio.2024.05.007
PMID:38784196
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11112005/
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/f4917db26ebb/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7762/11112005/0bf614b618af/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7762/11112005/effd4071064e/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7762/11112005/f4917db26ebb/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7762/11112005/0bf614b618af/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7762/11112005/effd4071064e/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7762/11112005/f4917db26ebb/gr4.jpg

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本文引用的文献

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Scaffold RNA engineering in type V CRISPR-Cas systems: a potent way to enhance gene expression in the yeast Saccharomyces cerevisiae.V 型 CRISPR-Cas 系统中的支架 RNA 工程:提高酵母酿酒酵母基因表达的有效方法。
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Logic Circuits Based on 2A Peptide Sequences in the Yeast .
基于酵母中2A肽序列的逻辑电路
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Engineering a two-gene system to operate as a highly sensitive biosensor or a sharp switch upon induction with β-estradiol.利用双基因系统,通过β-雌二醇诱导,实现高灵敏度生物传感器或锐利开关的功能。
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Synthetic metabolic transducers in Saccharomyces cerevisiae as sensors for aromatic permeant acids and bioreporters of hydrocarbon metabolism.酿酒酵母中的合成代谢传感器作为芳香渗透酸的传感器和烃代谢的生物报告器。
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Laccase production by Pleurotus ostreatus using cassava waste and its application in remediation of phenolic and polycyclic aromatic hydrocarbon-contaminated lignocellulosic biorefinery wastewater.利用木薯渣生产糙皮侧耳漆酶及其在修复含酚和多环芳烃的木质纤维素生物炼制废水中的应用。
Environ Pollut. 2022 Sep 15;309:119729. doi: 10.1016/j.envpol.2022.119729. Epub 2022 Jul 6.
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Synthetic Saccharomyces cerevisiae tolerate and degrade highly pollutant complex hydrocarbon mixture.合成酿酒酵母能耐受并降解高污染的复杂碳氢混合物。
Ecotoxicol Environ Saf. 2022 Aug;241:113768. doi: 10.1016/j.ecoenv.2022.113768. Epub 2022 Jun 17.
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Synthetic Transcriptional Networks Harnessing dCas12a and Type V-A anti-CRISPR Proteins.利用 dCas12a 和 V-A 型抗 CRISPR 蛋白构建人工转录网络。
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