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Title: Engineering Kluyveromyces marxianus as a Robust Synthetic Biology Platform Host

Journal Article · · mBio (Online)
 [1];  [1];  [2];  [3];  [4];  [5];  [6];  [7];  [8];  [1];  [9];  [10];  [9];  [11];
  1. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology
  2. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology; Univ. of California, Berkeley, CA (United States). Innovative Genomics Inst.
  3. Univ. of California, Berkeley, CA (United States). Dept. of Bioengineering; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division
  4. Energy Biosciences Inst., Berkeley, CA (United States)
  5. Univ. of California, Berkeley, CA (United States). Dept. of Bioengineering
  6. Univ. of California, Berkeley, CA (United States). Dept. of Chemistry
  7. Univ. of California, Berkeley, CA (United States). Dept. of Environmental Science, Policy, and Management
  8. Univ. of Campinas (UNICAMP), Sao Paulo (Brazil). Dept. of Genetics, Evolution and Bioagents
  9. Univ. of California, Berkeley, CA (United States). Dept. of Plant and Microbial Biology
  10. Univ. of California, Berkeley, CA (United States). Dept. of Bioengineering; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Biological Systems and Engineering Division; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Environmental Genomics and Systems Biology Division
  11. Univ. of California, Berkeley, CA (United States). Dept. of Molecular and Cell Biology; Univ. of California, Berkeley, CA (United States). Dept. of Chemistry; Lawrence Berkeley National Lab. (LBNL), Berkeley, CA (United States). Molecular Biophysics and Integrated Bioimaging
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Throughout history, the yeast Saccharomyces cerevisiae has played a central role in human society due to its use in food production and more recently as a major industrial and model microorganism, because of the many genetic and genomic tools available to probe its biology. However, S. cerevisiae has proven difficult to engineer to expand the carbon sources it can utilize, the products it can make, and the harsh conditions it can tolerate in industrial applications. Other yeasts that could solve many of these problems remain difficult to manipulate genetically. Here, we engineered the thermotolerant yeast Kluyveromyces marxianus to create a new synthetic biology platform. Using CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats with Cas9)-mediated genome editing, we show that wild isolates of K. marxianus can be made heterothallic for sexual crossing. By breeding two of these mating-type engineered K. marxianus strains, we combined three complex traits—thermotolerance, lipid production, and facile transformation with exogenous DNA—into a single host. The ability to cross K. marxianus strains with relative ease, together with CRISPR-Cas9 genome editing, should enable engineering of K. marxianus isolates with promising lipid production at temperatures far exceeding those of other fungi under development for industrial applications. These results establish K. marxianus as a synthetic biology platform comparable to S. cerevisiae, with naturally more robust traits that hold potential for the industrial production of renewable chemicals.

Research Organization:
Lawrence Berkeley National Laboratory (LBNL), Berkeley, CA (United States)
Sponsoring Organization:
USDOE Office of Science (SC)
Grant/Contract Number:
AC02-05CH11231
OSTI ID:
1626133
Journal Information:
mBio (Online), Vol. 9, Issue 5; ISSN 2150-7511
Publisher:
American Society for Microbiology (ASM)Copyright Statement
Country of Publication:
United States
Language:
English

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Cited By (5)

Stress-tolerant non-conventional microbes enable next-generation chemical biosynthesis journal January 2020
Early-branching gut fungi possess a large, comprehensive array of biomass-degrading enzymes journal February 2016
Blastobotrys adeninivorans and B. raffinosifermentans, two sibling yeast species which accumulate lipids at elevated temperatures and from diverse sugars journal June 2019
YEASTRACT+: a portal for cross-species comparative genomics of transcription regulation in yeasts journal October 2019
Engineering a CRISPR Interference System To Repress a Class 1 Integron in Escherichia coli journal February 2020

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59 BASIC BIOLOGICAL SCIENCES
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