Engineering of Yarrowia lipolytica transporters for high-efficient production of biobased succinic acid from glucose

Zhennan Jiang,Ziwei Zhu,Yinghang Liu,Qingsheng Qi,Ya-Jie Tang,Jin Hou,Zhiyong Cui

doi:10.1186/s13068-021-01996-w

Abstract

BackgroundSuccinic acid (SA) is a crucial metabolic intermediate and platform chemical. Development of biobased processes to achieve sustainable SA production has attracted more and more attention in biotechnology industry. Yarrowia lipolytica has a strong tricarboxylic acid cycle and tolerates low pH conditions, thus making it a potential platform for SA production. However, its SA titers in glucose media remain low.ResultsIn this study, we screened mitochondrial carriers and C4-dicarboxylic acid transporters to enhance SA secretion in Y. lipolytica. PGC62-SYF-Mae strain with efficient growth and SA production was constructed by optimizing SA biosynthetic pathways and expressing the transporter SpMae1. In fed-batch fermentation, this strain produced 101.4 g/L SA with a productivity of 0.70 g/L/h and a yield of 0.37 g/g glucose, which is the highest SA titer achieved using yeast, with glucose as the sole carbon resource.ConclusionOur results indicated that transporter engineering is a powerful strategy to achieve the efficient secretion of SA in Y. lipolytica, which will promote the industrial production of bio-based SA.

Highlights

Succinic acid (SA) is a crucial metabolic intermediate and platform chemical
The YlDic1 gene of the PGC62 strain was knocked down using the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) interference (CRISPRi, Additional file 1: Figure S1) method to obtain PGC62-YlDici, and the results were compared with those of PGC62 and PGC62-YlDic
0.57 ± 0.04 0.53 ± 0.01 0.62 ± 0.12 0.52 ± 0.05 0.59 ± 0.01 0.57 ± 0.03 illustrated in Fig. 2, the Cell dry weight (DCW) and SA titer of PGC62YlDici decreased by 58.9% and 26.4% compared with PGC62 strain, reached 4.2 g/L and 13.8 g/L, respectively

Summary

Introduction

Succinic acid (SA) is a crucial metabolic intermediate and platform chemical. Development of biobased processes to achieve sustainable SA production has attracted more and more attention in biotechnology industry. Yarrowia lipolytica has a strong tricarboxylic acid cycle and tolerates low pH conditions, making it a potential platform for SA production. Succinic acid (SA) is a vital C4 platform chemical, which has a wide range of applications in food and chemical engineering [1,2,3]. With the development of metabolic engineering and synthetic biology, this yeast has increasingly been applied to other biotechnological fields [10,11,12,13]. In 2010, Yuzbashev et al [14] inactivated succinate dehydrogenase (SDH) subunits and achieved SA accumulation through the oxidative TCA pathway in Y. lipolytica. Our group previously constructed an engineered Y. lipolytica strain named PGC202 with robust properties by deleting the SDH5 subunit encoding gene YlSdh and the acetyl-CoA hydrolase encoding gene YlAch, and overexpressing phosphoenolpyruvate carboxykinase (ScPck)

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Conclusion