Abstract
β-Arbutin is a plant-derived glycoside and widely used in cosmetic and pharmaceutical industries because of its safe and effective skin-lightening property as well as anti-oxidant, anti-microbial, and anti-inflammatory activities. In recent years, microbial fermentation has become a highly promising method for the production of β-arbutin. However, this method suffers from low titer and low yield, which has become the bottleneck for its widely industrial application. In this study, we used β-arbutin to demonstrate methods for improving yields for industrial-scale production in Escherichia coli. First, the supply of precursors phosphoenolpyruvate and uridine diphosphate glucose was improved, leading to a 4.6-fold increase in β-arbutin production in shaking flasks. The engineered strain produced 36.12 g/L β-arbutin with a yield of 0.11 g/g glucose in a 3-L bioreactor. Next, based on the substrate and product’s structural similarity, an endogenous O-acetyltransferase was identified as responsible for 6-O-acetylarbutin formation for the first time. Eliminating the formation of byproducts, including 6-O-acetylarbutin, tyrosine, and acetate, resulted in an engineered strain producing 43.79 g/L β-arbutin with a yield of 0.22 g/g glucose in fed-batch fermentation. Thus, the yield increased twofold by eliminating byproducts formation. To the best of our knowledge, this is the highest titer and yield of β-arbutin ever reported, paving the way for the industrial production of β-arbutin. This study demonstrated a systematic strategy to alleviate undesirable byproduct accumulation and improve the titer and yield of target products.
Key points
• A systematic strategy to improve titer and yield was showed
• Genes responsible for 6-O-acetylarbutin formation were firstly identified
• 43.79 g/L β-arbutin was produced in bioreactor, which is the highest titer so far
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Funding
This work was supported by the National Key Research and Development Program of China (2021YFC2100800), the National Natural Science Foundation of China (Grants 22078011 and 22238001) and Tianjin Synthetic Biotechnology Innovation Capacity Improvement Project (TSBICIP-KJGG-009).
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QY, XShen and NA conceived and designed the study. NA conducted the experiments and wrote the original manuscript. SZ and XC assisted to carry out the experiment. QY, XShen, XSun, JW and NA analyzed data. QY and XShen contributed to the review and editing of the manuscript. All authors read and approved the manuscript.
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An, N., Zhou, S., Chen, X. et al. High-yield production of β-arbutin by identifying and eliminating byproducts formation. Appl Microbiol Biotechnol 107, 6193–6204 (2023). https://doi.org/10.1007/s00253-023-12706-x
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DOI: https://doi.org/10.1007/s00253-023-12706-x