[논문]Optimized M9 Minimal Salts Medium for Enhanced Growth Rate and Glycogen Accumulation of Escherichia coli DH5α

Wang, Liang; Liu, Qinghua; Du, Yangguang; Tang, Daoquan; Wise, Michael J.

doi:10.4014/mbl.1804.04010

Optimized M9 Minimal Salts Medium for Enhanced Growth Rate and Glycogen Accumulation of Escherichia coli DH5α 원문보기

Microbiology and biotechnology letters = 한국미생물·생명공학회지, v.46 no.3, 2018년, pp.194 - 200

Wang, Liang (Department of Bioinformatics, School of Medical Informatics, Xuzhou Medical University) , Liu, Qinghua (Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, School of Pharmacy, Xuzhou Medical University) , Du, Yangguang (Xuzhou Center for Disease Control and Prevention) , Tang, Daoquan (Jiangsu Key Laboratory of New Drug Research and Clinical Pharmacy, School of Pharmacy, Xuzhou Medical University) , Wise, Michael J. (Computer Science and Software Engineering, University of Western Australia)

Abstract ▼ AI-Helper

Glycogen plays important roles in bacteria. Its structure and storage capability have received more attention recently because of the potential correlations with environmental durability and pathogenicity. However, the low level of intracellular glycogen makes extraction and structure characterization difficult, inhibiting functional studies. Bacteria grown in regular media such as lysogeny broth and tryptic soy broth do no accumulate large amounts of glycogen. Comparative analyses of bacterial media reported in literature for glycogen-related studies revealed that there was no consistency in the recipes reported. Escherichia coli $DH5{\alpha}$ is a convenient model organism for gene manipulation studies with respect to glycogen. Additionally, M9 minimal salts medium is widely used to improve glycogen accumulation, although its composition varies. In this study, we optimized the M9 medium by adjusting the concentrations of itrogen source, tryptone, carbon source, and glucose, in order to achieve a balance between the growth rate and glycogen accumulation. Our result showed that $1{\times}M9$ minimal salts medium containing 0.4% tryptone and 0.8% glucose was a well-balanced nutrient source for enhancing the growth and glycogen storage in bacteria. This result will help future investigations related to bacterial physiology in terms of glycogen function.

주제어

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문제 정의

A variety of media, including LB, Kornberg, MOPS, and M9 are used and the supplements have varied between studies. The initial purpose of our study was to investigate how Nterminus of glycogen branching enzyme (GBE) influences glycogen structure. Thus, we constructed a set of E.

제안 방법

All data obtained from experiments were analysed statistically. Excel and R packages were used for pairwised Student’s t-test and graph illustrations.

대상 데이터

Bacterial strains used in this study include E. coli BL21 (DE3), E. coli DH5α, E. coli RR1 and E. coli IK5.
We then tested the commercial M9 minimal salts (Sigma) for glycogen accumulation based on literature reports [5, 29]. The main ingredients of M9 minimal salts include Na₂HPO₄, KH₂PO₄, NaCl and NH₄Cl. However, E.
coli DH5α cultured in five types of 1 × M9 minimal salt media with different T/G ratio at 22 h. Three replicates were used for each medium. Glucose/Protein ratio (G/P) was used as an equivalent unit of glycogen concentration accumulated in E.

데이터처리

Excel and R packages were used for pairwised Student’s t-test and graph illustrations.

성능/효과

Significant differences were observed based on two-tailed student-t test for T/G = 1:8, T/G = 1:16, and T/G = 1:80 when they were compared with T/G = 1:2, respectively (*, p-value <0.05).
The results showed that optimized 1 × M9 minimal salt medium (T/G = 1:2) is sufficient for all strains to grow quickly and consistently and no significant differences were observed between the two media in terms of growth rates (Fig. 2).

후속연구

The observed bacterial clustering might be caused by bacterial auto-aggregation because of chemotaxis induced by over-nutrition[32]. However, specific mechanisms for the clustering require further investigation and are beyond the scope of this study. No similar phenomena were observed for other media.

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