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Related Experiment Video

Updated: Jul 16, 2026

Techniques for the Evolution of Robust Pentose-fermenting Yeast for Bioconversion of Lignocellulose to Ethanol

Techniques for the Evolution of Robust Pentose-fermenting Yeast for Bioconversion of Lignocellulose to Ethanol

Published on: October 24, 2016

Metabolism-driven and high-efficiency mining of ethanol-tolerant microorganisms from pit mud microbiota using

Teng Xu¹, Qing Sun², Gongchao Jing³

¹Single-Cell Center, Key Laboratory of Photoelectric Conversion and Utilization of Solar Energy, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, Shandong, China; University of Jinan, Jinan, Shandong, China.

Bioresource Technology

|July 14, 2026

Summary

We developed a high-throughput microfluidic optical tweezers-based Raman-activated cell sorting (RACS) system to efficiently find ethanol-tolerant microbes. This method significantly improves the discovery of stress-tolerant microorganisms compared to traditional techniques.

Keywords:

Ethanol tolerance Functional microbe mining Pit mud microbiome Raman-activated cell sorting Single-cell Raman spectroscopy Vitality

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Last Updated: Jul 16, 2026

Techniques for the Evolution of Robust Pentose-fermenting Yeast for Bioconversion of Lignocellulose to Ethanol

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Published on: October 24, 2016

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Published on: January 23, 2019

Area of Science:

Microbiology
Biotechnology
Spectroscopy

Background:

Identifying stress-tolerant microorganisms is crucial for developing robust microbial platforms.
Conventional culture-based methods for isolating these microbes are inefficient and labor-intensive.

Purpose of the Study:

To develop and apply a high-throughput system for efficient enrichment of stress-tolerant microorganisms.
To demonstrate the effectiveness of Raman-activated cell sorting (RACS) coupled with deuterium oxide (D2O)-labelled single-cell Raman spectroscopy (SCRS) for this purpose.

Main Methods:

A microfluidic optical tweezers-based RACS system was developed and integrated with D2O-labelled SCRS.
The system achieved a screening throughput of ~2,400 cells/h with 91.3% sorting accuracy.
Pit mud microbiomes were screened for ethanol-tolerant cells under stress conditions.

Main Results:

The D2O-RACS system successfully enriched 177 metabolically active, ethanol-tolerant cells.
Targeted cultivation yielded 6 isolates, all demonstrating strong tolerance to 8% ethanol, outperforming conventional screening (2/9 success).
Genome sequencing and transcriptomic profiling confirmed the ethanol-tolerant phenotypes of Lactiplantibacillus plantarum F4 and Staphylococcus epidermidis F5.

Conclusions:

The integrated D2O-RACS workflow offers a powerful and versatile platform for efficient mining of stress-tolerant microorganisms.
This screen-before-culture approach significantly improves enrichment (4.5-fold) and assessment efficiency (6.86-fold) over conventional methods.
The system enables rapid identification of high-performance microbial chassis for biotechnological applications.