A Combination of Flow Cytometry and Traditional Screening Using Chemicals to Isolate High Glutathione-Producing Yeast Mutants

Traditional screening using chemicals or flow cytometry (FCM) alone is not sufficient to isolate the high glutathione (GSH)-producing yeast strains used in food production. Therefore, to improve screening efficiency, we investigated a combination of both methods. A mutated Saccharomyces cerevisiae s...

Full description

Saved in:
Bibliographic Details
Published in:Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2012, Vol.76 (6), p.1085-1090
Main Authors: NISHIUCHI, Hiroaki, TABIRA, Yukiko, YAMAGISHI, Kazuo
Format: Article
Language:English
Subjects:
5-chloromethylfluorescein diacetate (CMFDA)
Biological and medical sciences
Cerulenin - pharmacology
efficient screening method
Fatty Acid Synthesis Inhibitors - pharmacology
Flow Cytometry
Fluoresceins
Fluorescent Dyes
Food Technology
Fundamental and applied biological sciences. Psychology
glutathione (GSH)-producing yeast
Glutathione - biosynthesis
Glutathione - genetics
High-Throughput Screening Assays
Mutation
Saccharomyces cerevisiae
Saccharomyces cerevisiae - drug effects
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - isolation & purification
Saccharomyces cerevisiae - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Traditional screening using chemicals or flow cytometry (FCM) alone is not sufficient to isolate the high glutathione (GSH)-producing yeast strains used in food production. Therefore, to improve screening efficiency, we investigated a combination of both methods. A mutated Saccharomyces cerevisiae strain was labeled with 5-chloromethylfluorescein diacetate and sorted by FCM according to emitted fluorescence intensity. Moderate GSH (1%-2%)-producing mutants were isolated, whereas high GSH (>2%)-producing mutants were not. Traditional screening using cerulenin resulted in similar findings, but a combination of both methods resulted in a 40% increase in the screening yield of high GSH-producing mutants. An analysis of model strains indicated that the ratio of high GSH-producing cells in a sample affected the FCM results. By combining FCM with traditional screening using chemicals, we succeeded in isolating high GSH-producing mutants from several parental strains.
ISSN:0916-8451
1347-6947
DOI:10.1271/bbb.110883