Optimization of the Culture Medium for Efficient Nervonic Acid Production by Engineered Rhodosporidium toruloides

IF 2.4 4区 农林科学 Q3 CHEMISTRY, APPLIED
Feixiang Liu, Chenan Jiang, Tingting Lu, Xiaoxin Liu, Erzheng Su
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Abstract

Nervonic acid (NA), a very long-chain monounsaturated fatty acid with critical roles in neurological health and brain development, has garnered increasing interest for microbial production via metabolic engineering. Previously, the laboratory constructed a recombinant strain using Rhodosporidium toruloides as the chassis, which is capable of synthesizing NA. However, the lack of systematic medium optimization remains a major bottleneck for enhancing NA titers in the engineered strains. This study aimed to develop an economical and efficient fermentation medium for NA biosynthesis using the engineered strain in shake-flask cultures. Through one-factor-at-a-time optimization, glucose and corn steep liquor (CSL) were identified as the optimal carbon and organic nitrogen sources, respectively. Further analysis revealed that precise concentrations of glucose (120 g/L), CSL (10 g/L), and ammonium sulfate (1.1 g/L) were critical for balancing cell growth and lipogenesis. Notably, inorganic salts (CaCl2, MgSO4·7H2O, KH2PO4) were dispensable in the presence of CSL, which inherently supplied essential trace elements. Maintaining the initial pH at its natural value (4.3) proved optimal, as artificial pH adjustments severely impaired lipid accumulation. Under the optimized conditions, the NA titer reached 7.5 g/L, representing a 29.3% increase over the initial medium (5.8 g/L) and establishing the highest reported shake-flask titer for microbial NA production. This work not only provides a cost-effective medium formulation by eliminating inorganic salts and utilizing low-cost CSL but also offers strategic insights for scaling up NA biosynthesis in bioreactors. The findings underscore the importance of tailored nutrient optimization in aligning metabolic engineering advancements with bioprocess practicality.

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工程红孢子虫高效产神经酸培养基的优化
神经酸(NA)是一种长链单不饱和脂肪酸,在神经系统健康和大脑发育中起着至关重要的作用,通过代谢工程已经引起了微生物生产越来越多的兴趣。此前,实验室以环形Rhodosporidium toruloides为底盘构建了一株能够合成NA的重组菌株。然而,缺乏系统的培养基优化仍然是提高工程菌株NA滴度的主要瓶颈。本研究旨在开发一种经济高效的发酵培养基,利用该工程菌株在摇瓶培养中合成NA。通过单因素优化,确定葡萄糖和玉米浸泡液分别为最佳碳源和有机氮源。进一步分析表明,葡萄糖(120 g/L)、CSL (10 g/L)和硫酸铵(1.1 g/L)的精确浓度对平衡细胞生长和脂肪生成至关重要。值得注意的是,无机盐(CaCl2, MgSO4·7H2O, KH2PO4)在CSL存在下是不可缺少的,它们本身提供必需的微量元素。将初始pH值维持在自然值(4.3)被证明是最佳的,因为人工调节pH值严重损害了脂质积累。在优化条件下,NA滴度达到7.5 g/L,比初始培养基(5.8 g/L)提高了29.3%,建立了报道的微生物NA生产的最高摇瓶滴度。这项工作不仅通过消除无机盐和利用低成本的CSL提供了一种具有成本效益的培养基配方,而且为扩大生物反应器中NA的生物合成提供了战略见解。这些发现强调了量身定制的营养优化在使代谢工程进步与生物过程实用性相结合方面的重要性。
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来源期刊
CiteScore
4.10
自引率
5.00%
发文量
95
审稿时长
2.4 months
期刊介绍: The Journal of the American Oil Chemists’ Society (JAOCS) is an international peer-reviewed journal that publishes significant original scientific research and technological advances on fats, oils, oilseed proteins, and related materials through original research articles, invited reviews, short communications, and letters to the editor. We seek to publish reports that will significantly advance scientific understanding through hypothesis driven research, innovations, and important new information pertaining to analysis, properties, processing, products, and applications of these food and industrial resources. Breakthroughs in food science and technology, biotechnology (including genomics, biomechanisms, biocatalysis and bioprocessing), and industrial products and applications are particularly appropriate. JAOCS also considers reports on the lipid composition of new, unique, and traditional sources of lipids that definitively address a research hypothesis and advances scientific understanding. However, the genus and species of the source must be verified by appropriate means of classification. In addition, the GPS location of the harvested materials and seed or vegetative samples should be deposited in an accredited germplasm repository. Compositional data suitable for Original Research Articles must embody replicated estimate of tissue constituents, such as oil, protein, carbohydrate, fatty acid, phospholipid, tocopherol, sterol, and carotenoid compositions. Other components unique to the specific plant or animal source may be reported. Furthermore, lipid composition papers should incorporate elements of year­to­year, environmental, and/ or cultivar variations through use of appropriate statistical analyses.
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