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Shane Bassett¹, Jonathan C Suganda¹, Nancy A Da Silva¹

¹Department of Chemical & Biomolecular Engineering, University of California, Irvine, CA, 92697-2580, USA.

Metabolic engineering

|November 3, 2024

概括

我们设计了酵母Kluyveromyces marxianus用于过氧体表面显示,增强了工业生物制造. 这种方法通过提高酶同位化和途径效率来促进有价值化合物的产生.

科学领域:

合成生物学合成生物学
生物技术是生物技术.
微生物工程微生物工程

背景情况:

克鲁伊沃罗密塞斯马克西安纳斯 (Kluyveromyces marxianus) 是工业生物制造中的一种有价值的酵母.
基因组编辑和合成生物学工具的进步增加了K. marxianus的工程可访问性.
酶局部化策略增强了代谢途径的流动和非原生化合物的产生.

研究的目的:

为了设计K. marxianus以使用Pex15 anchoring motif进行过氧体表面显示.
为了证明过氧体表面显示器在改善各种非本地产品的生产方面的实用性.
提高K. marxianus.的工业生物制造能力.

主要方法:

识别并利用了本土的Kluyveromyces marxianus Pex15 (KmPex15) anchoring motif. 发现并利用了本土的Kluyveromyces marxianus Pex15 (KmPex15) anchoring motif. 发现并利用了本土的Kluyveromyces marxianus Pex15 (KmPex15) anchoring motif. 发现并利用了本土的Kluyveromyces marxianus Pex15 (KmPex15) anchoring motif.
融合了KmPex15基因与记者蛋白 (例如GFP) 以及氧体表面显示的通路酶.
工程化过氧体形态和寿命,以进一步优化生产.

主要成果:

在K. marxianus.成功地将GFP局部化到过氧体表面.
通过同时显示 Pseudomonas savastanoi IaaM 和 IaaH 酶,实现了 7.9 倍的英多尔-3 酸生产.
通过使用Chromobacterium violaceum VioE和VioD酶,提高了proviolacein与prodeoxyviolacein的选择性2.5倍.
通过同时显示Cat2,Acc1和2-pyrone合成酶在过氧体表面上,增加了三酸乳 (TAL) 位数的2倍.
通过过氧体工程,进一步提高了TAL产量的2.1倍.

结论:

过氧体表面显示是一种有效的酶局部化策略,在K. marxianus.
这种方法显著提高了各种非本地产品的生产.
工程化K. marxianus平台为工业生物制造提供了改进的能力.

关键词:

乙-甲酸是一种酶局部化酶的局部化.克鲁伊维罗米塞斯马克西安纳斯 (Kluyveromyces marxianus) 是一个植物.在PEX15中使用PEX15 过氧体表面显示显示器

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