Affiliation(s): 1Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310058, China
2Ningbo Innovation Center, Zhejiang University, Ningbo 315100, China
Corresponding email(s):
Mianbin WU, wumb@zju.edu.cn
Lidan YE, yelidan@zju.edu.cn
Abstract: Industrial biomanufacturing of D-arabitol, a high-value sugar alcohol, is currently constrained by the scarcity of robust and high-yielding microbial strains. To address this issue, we isolated a wild-type strain, Wickerhamomyces anomalus K955, and engineered it using a combinatorial mutagenesis strategy that integrated low-energy N+ ion implantation with diethyl sulfate (DES) treatment, coupled with an iodonitrotetrazolium violet (INT)-based high-throughput screening (HTS) method. The resulting mutant, A37, exhibited superior performance, achieving a titer of 108.69 g/L and a yield of 0.31 g/g in shake-flask fermentation. Transcriptomic analysis showed that this hyper-producing phenotype resulted from global metabolic reprogramming: the strain demonstrated enhanced biosynthetic capacity via upregulated ribosome biogenesis, while carbon flux was effectively redirected toward the Pentose Phosphate Pathway (PPP) through synergistic upregulation of key genes, including zwf, gnd, and rpe. In scaled-up fed-batch fermentation, strain A37 produced 220.85 g/L of D-arabitol with a productivity of 1.28 g/(L·h), representing the highest levels reported to date. These findings provide a superior candidate strain for industrial D-arabitol production and offer critical insights into the metabolic regulation mechanisms underlying high-efficiency biosynthesis.
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