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从木糖母液中逐步代谢工程化热带假丝酵母生产高效木糖醇。

Stepwise metabolic engineering of Candida tropicalis for efficient xylitol production from xylose mother liquor.

机构信息

Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, People's Republic of China.

出版信息

Microb Cell Fact. 2021 May 25;20(1):105. doi: 10.1186/s12934-021-01596-1.

Abstract

BACKGROUND

Commercial xylose purification produces xylose mother liquor (XML) as a major byproduct, which has become an inexpensive and abundant carbon source. A portion of this XML has been used to produce low-value-added products such as caramel but the remainder often ends up as an organic pollutant. This has become an issue of industrial concern. In this study, a uracil-deficient Candida tropicalis strain was engineered to efficiently convert XML to the commercially useful product xylitol.

RESULTS

The xylitol dehydrogenase gene was deleted to block the conversion of xylitol to xylulose. Then, an NADPH regeneration system was added through heterologous expression of the Yarrowia lipolytica genes encoding 6-phosphate-gluconic acid dehydrogenase and 6-phosphate-glucose dehydrogenase. After process optimization, the engineered strain, C. tropicalis XZX-B4ZG, produced 97.10 g L xylitol in 120 h from 300 g L XML in a 5-L fermenter. The xylitol production rate was 0.82 g Lh and the conversion rate was 92.40 %.

CONCLUSIONS

In conclusion, this study performed a combination of metabolic engineering and process optimizing in C. tropicalis to enhance xylitol production from XML. The use of C. tropicalis XZX-B4ZG, therefore, provided a convenient method to transform the industrial by-product XML into the useful material xylitol.

摘要

背景

商业木糖提纯会产生木糖母液(XML)作为主要副产物,这已成为一种廉价且丰富的碳源。一部分 XML 已被用于生产焦糖等低附加值产品,但其余部分通常最终成为有机污染物。这已成为工业关注的问题。在这项研究中,通过工程改造缺乏尿嘧啶的热带假丝酵母菌株,使其能够有效地将 XML 转化为具有商业价值的产品木糖醇。

结果

删除木糖醇脱氢酶基因以阻止木糖醇转化为木酮糖。然后,通过异源表达解脂耶氏酵母的基因编码 6-磷酸葡萄糖酸脱氢酶和 6-磷酸葡萄糖脱氢酶来添加 NADPH 再生系统。经过工艺优化,工程菌株 C. tropicalis XZX-B4ZG 在 5-L 发酵罐中从 300 g/L XML 中生产 97.10 g/L 木糖醇,历时 120 h。木糖醇的生产速率为 0.82 g/L/h,转化率为 92.40%。

结论

总之,本研究在热带假丝酵母中进行了代谢工程和工艺优化的组合,以提高 XML 生产木糖醇的能力。因此,使用 C. tropicalis XZX-B4ZG 为将工业副产物 XML 转化为有用物质木糖醇提供了一种便捷的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/fa49/8147403/80e7dda972bb/12934_2021_1596_Fig1_HTML.jpg

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