Development of Corynebacterium glutamicum as a monoterpene production platform

• DOI: 10.1016/j.ymben.2023.11.009
• Publication Date: 2023-12-04T16:58:24Z
• Authors: Bridget A. Luckie; Meera Kashyap; Allison N. Pearson; Yan Chen; Yuzhong Liu; Luis E. Valencia; Alexander Carrill...; Graham A. Hudson; Xavier B. Tao; Bryan Wu; Christopher J. Pe...; Jay D. Keasling
• Keywords: Corynebacterium glutamicum; Monoterpene; Bioproduction; Terpene

Monoterpenes are commonly known for their role in the flavors and fragrances industry also gaining attention other uses like insect repellant as potential renewable fuels aviation. Corynebacterium glutamicum, a Generally Recognized Safe microbe, has been choice organism annual million ton-scale bioproduction of amino acids more than 50 years; however, efforts to produce monoterpenes C. glutamicum have remained relatively limited. In this study, we report further expansion biosynthetic repertoire through development optimization mevalonate-based monoterpene platform. course our plasmid design iterations, increased flux bypass pathway, measuring isoprenol production proxy precursor abundance demonstrating highest reported titers date at 1504.6 mg/L. Our designs evaluated effects backbone, promoter, GPP synthase homolog origin on product titers. Monoterpene was improved by disrupting competing pathways isoprenoid supply implementing biphasic system prevent volatilization. With platform, achieved 321.1 mg/L geranoids, 723.6 1,8-cineole, 227.8 linalool. Furthermore, determined that first oxidizes geraniol an aldehyde intermediate before it is asymmetrically reduced citronellol. Additionally, demonstrate reductase, AdhC, possesses additional substrate promiscuity acyclic aldehydes.