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  • P-ISSN2233-4203
  • E-ISSN2093-8950
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  • P-ISSN 2233-4203
  • E-ISSN 2093-8950

Relative Quantification of Glycans by Metabolic Isotope Labeling with Isotope Glucose in Aspergillus niger

Relative Quantification of Glycans by Metabolic Isotope Labeling with Isotope Glucose in Aspergillus niger

Mass Spectrometry Letters / Mass Spectrometry Letters, (P)2233-4203; (E)2093-8950
2022, v.13 no.4, pp.139-145
https://doi.org/10.5478/MSL.2022.13.4.139
Soo-Hyun Choi (Changwon National University)
Ye-Eun Cho (Changwon National University)
Do-Hyun Kim (Changwon National University)
Jin-il Kim (Changwon National University)
Jihee Yun (Changwon National University)
Jae-Yoon Jo (Changwon National University)
Jae-Min Lim (Changwon National University)
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초록

Protein glycosylation is a common post-translational modification by non-template-based biosynthesis. In fungal biotechnology, which has great applications in pharmaceuticals and industries, the importance of research on fungal glycopro- teins and glycans is accelerating. In particular, the importance of quantitative analysis of fungal glycans is emerging in research on the production of filamentous fungal proteins by genetic modification. Reliable mass spectrometry-based techniques for quantitative glycomics have evolved into chemical, enzymatic, and metabolic stable isotope labeling methods. In this study, we intend to expand quantitative glycomics by metabolic isotope labeling of glycans in Aspergillus niger, a filamentous fungus model, by the MILPIG method. We demonstrate that incubation of filamentous fungi in a culture medium with carbon-13 labeled glucose (1- 13 C 1 ) efficiently incorporates carbon-13 into N-linked glycans. In addition, for quantitative validation of this method, light and heavy glycans are mixed 1:1 to show the performance of quantitative analysis of various N-linked glycans simultaneously. We have successfully quantified fungal glycans by MILPIG and expect it to be widely applicable to glycan expression levels under various biological conditions in fungi.

keywords
Aspergillus niger, Glycans, Glycosylation, Metabolic labeling, Isotopic glucose, Quantitative glycomics, MILPIG, Mass spectrometry

Abstract

Protein glycosylation is a common post-translational modification by non-template-based biosynthesis. In fungal biotechnology, which has great applications in pharmaceuticals and industries, the importance of research on fungal glycopro- teins and glycans is accelerating. In particular, the importance of quantitative analysis of fungal glycans is emerging in research on the production of filamentous fungal proteins by genetic modification. Reliable mass spectrometry-based techniques for quantitative glycomics have evolved into chemical, enzymatic, and metabolic stable isotope labeling methods. In this study, we intend to expand quantitative glycomics by metabolic isotope labeling of glycans in Aspergillus niger, a filamentous fungus model, by the MILPIG method. We demonstrate that incubation of filamentous fungi in a culture medium with carbon-13 labeled glucose (1- 13 C 1 ) efficiently incorporates carbon-13 into N-linked glycans. In addition, for quantitative validation of this method, light and heavy glycans are mixed 1:1 to show the performance of quantitative analysis of various N-linked glycans simultaneously. We have successfully quantified fungal glycans by MILPIG and expect it to be widely applicable to glycan expression levels under various biological conditions in fungi.

keywords
Aspergillus niger, Glycans, Glycosylation, Metabolic labeling, Isotopic glucose, Quantitative glycomics, MILPIG, Mass spectrometry


투고일Submission Date
2022-11-27
수정일Revised Date
2022-12-12
게재확정일Accepted Date
2022-12-14
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