Developing a broad-range promoter set for metabolic engineering in the thermotolerant yeast Kluyveromyces marxianus
| Autor: | Nancy A. Da Silva, Mengwan Li, Pamela B. Besada-Lombana, Ian Wheeldon, Xuye Lang |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
0106 biological sciences
Endocrinology Diabetes and Metabolism lcsh:Biotechnology Saccharomyces cerevisiae Biomedical Engineering Xylose 01 natural sciences Metabolic engineering 03 medical and health sciences chemistry.chemical_compound Kluyveromyces marxianus Special issue on Non-conventional microbes edited by Ian Wheeldon and Aindrila Mukhopadhyay Thermo-tolerance 010608 biotechnology lcsh:TP248.13-248.65 K. marxianus lcsh:QH301-705.5 030304 developmental biology Triacetic acid lactone 0303 health sciences biology Promoter biology.organism_classification Yeast Citric acid cycle chemistry Biochemistry lcsh:Biology (General) Non-conventional microbe Chemical production |
| Zdroj: | Metabolic Engineering Communications Metabolic Engineering Communications, Vol 11, Iss, Pp e00145-(2020) |
| ISSN: | 2214-0301 |
| Popis: | Kluyveromyces marxianus is an emerging host for metabolic engineering. This thermotolerant yeast is the fastest growing eukaryote, has high flux through the TCA cycle, and can metabolize a broad range of C5, C6, and C12 carbon sources. In comparison to the common host Saccharomyces cerevisiae, this non-conventional yeast suffers from a lack of metabolic engineering tools to control gene expression over a wide transcriptional range. To address this issue, we designed a library of 25 native-derived promoters from K. marxanius CBS6556 that spans 87-fold transcriptional strength under glucose metabolism. Six promoters from the library were further characterized in both glucose and xylose as well as across various temperatures from 30 to 45 °C. The temperature study revealed that in most cases EGFP expression decreased with elevating temperature; however, two promoters, PSSA3 and PADH1, increased expression above 40 °C in both xylose and glucose. The six-promoter set was also validated in xylose for triacetic acid lactone (TAL) production. By controlling the expression level of heterologous 2-pyrone synthase (2-PS), the specific TAL titer increased over 8-fold at 37 °C. Cultures at 41 °C exhibited a similar TAL biosynthesis capability, while at 30 °C TAL levels were lower. Taken together, these results advance the metabolic engineering tool set in K. marxianus and further develop this new host for chemical biosynthesis. Highlights • Library of 25 K. marxianus promoters spans nearly 2 order magnitude of transcriptional strength under glucose metabolism. • Promoters show similar strength trends with both glucose and xylose metabolism. • KmNC1 promoter shows highest strength among reported K. marxianus native promoters. • KmNC1 promoter increased specific triacetic acid lactone (TAL) titer by 80% compared with previous studies. |
| Databáze: | OpenAIRE |
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