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Using gel microdroplets to develop a simple high-throughput screening platform for oleaginous microorganisms.

Yuma Tanaka1, Akihiro Nakamura2, Yoshiyuki Suzuki2

  • 1Department of Bioengineering, Nagaoka University of Technology, 1603-1 Kamitomioka, Nagaoka, Niigata 940-2188, Japan.

Journal of Biotechnology
|August 30, 2022
PubMed
Summary

Researchers developed a high-throughput method using gel microdroplets (GMD) to screen for slow-growing mutants of the oleaginous yeast Lipomyces starkeyi, identifying a mutant with significantly delayed growth for further genetic study.

Keywords:
Gel microdropletsHigh-throughput screeningOleaginous yeast

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Area of Science:

  • Biotechnology
  • Microbiology
  • Synthetic Biology

Background:

  • Oleaginous yeast, like Lipomyces starkeyi, are promising sources for lipid production, accumulating over 85% lipid per dry cell weight.
  • Current screening methods for high-lipid mutants are laborious and hinder the evaluation of individual mutant growth.
  • Efficient screening for slow-growing mutants is crucial for understanding yeast growth regulation and improving lipid yields.

Purpose of the Study:

  • To develop a high-throughput screening method for evaluating both growth and lipid production in individual yeast mutants.
  • To identify slow-growing mutants of Lipomyces starkeyi with potential for novel genetic insights.
  • To enable simultaneous assessment of growth and lipid accumulation in yeast.

Main Methods:

  • Development of a high-throughput screening platform utilizing gel microdroplets (GMD).
  • Encapsulation of individual Lipomyces starkeyi mutants within GMD for simultaneous growth and lipid analysis.
  • Evaluation of microcolony growth by scattered light measurement within GMD.

Main Results:

  • Demonstrated successful growth of Lipomyces starkeyi within GMD, with growth assessed by scattered light.
  • Successfully identified a mutant exhibiting a 10-fold reduction in growth rate compared to the wild-type.
  • Established a method for simultaneous evaluation of yeast growth and lipid production.

Conclusions:

  • Gel microdroplets (GMD) provide an effective high-throughput platform for screening oleaginous yeast mutants.
  • The identified slow-growing mutant offers a valuable resource for investigating genes critical to yeast growth.
  • This method facilitates the discovery of mutants with altered growth characteristics, aiding in the optimization of lipid production in oleaginous yeasts.