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提高工业黑曲霉 CGMCC 10142 的柠檬酸产量:用不同启动子鉴定和过表达高亲和力葡萄糖转运蛋白。

Improving citric acid production of an industrial Aspergillus niger CGMCC 10142: identification and overexpression of a high-affinity glucose transporter with different promoters.

机构信息

Key Laboratory of Industrial Microbiology & Engineering Research Center of Food Biotechnology of Ministry of Education, College of Biotechnology, Tianjin University of Science and Technology, Tianjin, 300457, People's Republic of China.

Tianjin Key Laboratory of Industrial Fermentation Microbiology, Tianjin, 300457, People's Republic of China.

出版信息

Microb Cell Fact. 2021 Aug 26;20(1):168. doi: 10.1186/s12934-021-01659-3.

Abstract

BACKGROUND

Glucose transporters play an important role in the fermentation of citric acid. In this study, a high-affinity glucose transporter (HGT1) was identified and overexpressed in the industrial strain A. niger CGMCC 10142. HGT1-overexpressing strains using the PglaA and Paox1 promoters were constructed to verify the glucose transporter functions.

RESULT

As hypothesized, the HGT1-overexpressing strains showed higher citric acid production and lower residual sugar contents. The best-performing strain A. niger 20-15 exhibited a reduction of the total sugar content and residual reducing sugars by 16.5 and 44.7%, while the final citric acid production was significantly increased to 174.1 g/L, representing a 7.3% increase compared to A. niger CGMCC 10142. Measurement of the mRNA expression levels of relevant genes at different time-points during the fermentation indicated that in addition to HGT1, citrate synthase and glucokinase were also expressed at higher levels in the overexpression strains.

CONCLUSION

The results indicate that HGT1 overexpression resolved the metabolic bottleneck caused by insufficient sugar transport and thereby improved the sugar utilization rate. This study demonstrates the usefulness of the high-affinity glucose transporter HGT1 for improving the citric acid fermentation process of Aspergillus niger CGMCC 10142.

摘要

背景

葡萄糖转运蛋白在柠檬酸发酵中起着重要作用。在本研究中,鉴定并在工业菌株黑曲霉 CGMCC 10142 中过表达了一种高亲和力葡萄糖转运蛋白(HGT1)。使用 PglaA 和 Paox1 启动子构建了 HGT1 过表达株,以验证葡萄糖转运蛋白的功能。

结果

正如假设的那样,HGT1 过表达株表现出更高的柠檬酸产量和更低的残留糖含量。表现最好的菌株黑曲霉 20-15 将总糖含量和还原糖残留量分别降低了 16.5%和 44.7%,而最终柠檬酸产量显著增加到 174.1 g/L,比黑曲霉 CGMCC 10142 提高了 7.3%。在发酵过程中不同时间点测量相关基因的 mRNA 表达水平表明,除了 HGT1 外,柠檬酸合酶和葡萄糖激酶在过表达株中的表达水平也更高。

结论

结果表明,HGT1 的过表达解决了由于糖转运不足而导致的代谢瓶颈,从而提高了糖的利用率。本研究证明了高亲和力葡萄糖转运蛋白 HGT1 可用于改善黑曲霉 CGMCC 10142 的柠檬酸发酵过程。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/75c8/8394697/a9562c6aa136/12934_2021_1659_Fig1_HTML.jpg

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