• Mass Spectrometry Letters
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• v.13 no.4
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• pp.139-145
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• 2022

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 glycoproteins 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 (1-¹³C₁) 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.

• Journal of Nutrition and Health
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• v.22 no.1
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• pp.1-8
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• 1989

The study was attempted to observe effects of voluntary exercise on energy expenditure and on storage of excessive energy into body fat in adult mice. Mice, body weight 25-30g, were divided into two groups, exercise and sedentary group. Voluntary exercize by running wheel was allowed for former group but not for the mice of the latter group which were restricted by limiting space allowance 13.5$\times$11.5$\times$15.0cm per mouse. During a period of 4 weeks of feeding trial, they were fed ad libitum starch-casein based diet added with $^3\textrm{H}$-glucose (D-[1-$^3\textrm{H}$(N)]-glucose) at a level of 20 nCi per g of diet. Measurements were made to study hematology, lipase activity in epididymal adipose tissue, total contents of body waste and fat, and radiactivity of $^3\textrm{H}$-glucose incoporated into body fat. Dietary intake, body weight gain and amount of voluntary physical activities were also measured. The results obtained ard summarized as follows; 1) Amount of metabolizable energy intake, body weight gain and body fat were not statistically different between the two group. However, mice of the exercise group tended to show lower body weight gain body fat contants, but higher energy expenditure than those of the sedentary group. 2) Radioactivity of $^3\textrm{H}$-glucose incoporated into body fat appeared lower for exercise group expressd on a whole body fat basis. The activity was, however, higher for exercise group when expressed based on per g of fat compared to that of sedentary group. 3) Exercise group showed also higher activity of lipase in epididymal adipose tissue than the other group. 4) Mice whose physical activities were restricted appeared to have lower levels of hemoglobin and hematocrit values than of the exercise group. These results seem to support the theory that turn-over rate of body fat is activated by exercise and to suggest that consumed energy is to be converted primarily into body fat before its use as energy source by oxidation even during a period of continuous energy expenditure by exercise.