Jiaqi Mao , Huan Fang , Guangqing Du , Dawei Zhang
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引用次数: 0
摘要
生物素对三大营养素的代谢至关重要,对动物的生长和发育也至关重要。在工业上,生物素的生产主要依靠化学合成,这种方法存在工艺复杂、环境风险大和成本高等缺点。因此,研究人员越来越认识到生物合成生物素的好处。原生转录调控和代谢瓶颈限制了生物素的生产。用组成型 P43 启动子取代原生启动子和 bioO,并将原生 birA 换成结核分枝杆菌的非调控变体,可显著提高生物素生物合成基因的表达,有效绕过原生转录调控。绕过低效原生生物素的后续策略包括敲除β-氧化途径中的ydbM--一种潜在的嘧啶酰-CoA降解剂、表达异源生物素基因以及引入替代的嘧啶酰-ACP生物合成途径,这些策略共同将生物素滴度提高到了6.91毫克/升。这项研究展示了一种克服代谢和监管障碍的多方面方法,为提高枯草芽孢杆菌的生物素产量提供了一个模板,对更广泛的微生物生物技术领域具有重要意义。
Enhancing biotin production in Bacillus subtilis: Overcoming native pathway limitations
Biotin is essential for metabolizing the three major nutrients and is vital for animal growth and development. Industrially, biotin production has relied on chemical synthesis, a method fraught with disadvantages including complex processes, environmental risks, and high costs. Consequently, researchers have increasingly recognized the benefits of biosynthesizing biotin. Native transcriptional regulation and metabolic bottlenecks restrict biotin production. Replacing the native promoter and bioO with the constitutive P43 promoter, along with swapping the native birA for a non-regulatory variant from Mycobacterium tuberculosis, significantly upregulated biotin biosynthetic gene expression, effectively bypassing native transcriptional regulation. Subsequent strategies to bypass the inefficient native BioW included knocking out ydbM in the β-oxidation pathway—a potential pimeloyl-CoA degrader, expressing heterologous bioW genes, and introducing alternative pimeloyl-ACP biosynthetic pathways, collectively increasing biotin titers to 6.91 mg/L. This study demonstrates a multi-faceted approach to overcoming metabolic and regulatory barriers, providing a template for enhancing biotin production in B. subtilis, with implications for the broader field of microbial biotechnology.
期刊介绍:
Process Biochemistry is an application-orientated research journal devoted to reporting advances with originality and novelty, in the science and technology of the processes involving bioactive molecules and living organisms. These processes concern the production of useful metabolites or materials, or the removal of toxic compounds using tools and methods of current biology and engineering. Its main areas of interest include novel bioprocesses and enabling technologies (such as nanobiotechnology, tissue engineering, directed evolution, metabolic engineering, systems biology, and synthetic biology) applicable in food (nutraceutical), healthcare (medical, pharmaceutical, cosmetic), energy (biofuels), environmental, and biorefinery industries and their underlying biological and engineering principles.