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细菌血红蛋白 VHb 的异源过表达提高了酵母解脂耶氏酵母的赤藓糖醇生物合成。

Heterologous overexpression of bacterial hemoglobin VHb improves erythritol biosynthesis by yeast Yarrowia lipolytica.

机构信息

Department of Biotechnology and Food Microbiology, Wroclaw University of Environmental and Life Sciences, Chełmońskiego 37, 51-630, Wrocław, Poland.

出版信息

Microb Cell Fact. 2019 Oct 15;18(1):176. doi: 10.1186/s12934-019-1231-9.

Abstract

BACKGROUND

Yarrowia lipolytica is an unconventional yeast with a huge industrial potential. Despite many advantages for biotechnological applications, it possesses enormous demand for oxygen, which is a bottleneck in large scale production. In this study a codon optimized bacterial hemoglobin from Vitreoscilla stercoraria (VHb) was overexpressed in Y. lipolytica for efficient growth and erythritol synthesis from glycerol in low-oxygen conditions. Erythritol is a natural sweetener produced by Y. lipolytica under high osmotic pressure and at low pH, and this process requires high oxygen demand.

RESULTS

Under these conditions the VHb overexpressing strain showed mostly yeast-type cells resulting in 83% higher erythritol titer in shake-flask experiments. During a bioreactor study the engineered strain showed higher erythritol productivity (Q = 0.38 g/l h) and yield (Y = 0.37 g/g) in comparison to the control strain (Q = 0.30 g/l h, Y = 0.29 g/g). Moreover, low stirring during the fermentation process resulted in modest foam formation.

CONCLUSIONS

This study showed that overexpression of VHb in Y. lipolytica allows for dynamic growth and efficient production of a value-added product from a low-value substrate.

摘要

背景

解脂耶氏酵母是一种具有巨大工业潜力的非常规酵母。尽管在生物技术应用方面有许多优势,但它对氧气的需求巨大,这是大规模生产的一个瓶颈。在这项研究中,来自威氏气单胞菌的密码子优化细菌血红蛋白(VHb)在解脂耶氏酵母中过表达,以在低氧条件下从甘油高效生长和合成赤藓糖醇。赤藓糖醇是解脂耶氏酵母在高渗透压和低 pH 值下产生的天然甜味剂,这个过程需要高氧气需求。

结果

在这些条件下,过表达 VHb 的菌株主要表现为酵母型细胞,导致摇瓶实验中赤藓糖醇产量提高 83%。在生物反应器研究中,与对照菌株(Q=0.30 g/l/h,Y=0.29 g/g)相比,工程菌株表现出更高的赤藓糖醇生产力(Q=0.38 g/l/h)和产率(Y=0.37 g/g)。此外,发酵过程中低搅拌导致适度的泡沫形成。

结论

本研究表明,在解脂耶氏酵母中过表达 VHb 可以实现动态生长,并从低价值底物高效生产有价值的产品。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b561/6794898/4300c7289c4e/12934_2019_1231_Fig1_HTML.jpg

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