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Structural and physiological studies on the storage β-polyglucan of haptophyte Pleurochrysis haptonemofera

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Abstract

The storage β-polyglucan and catabolic enzyme activities of the haptophyte Pleurochrysis haptonemofera were characterized. The storage β-polyglucan was prepared by the dimethylsulfoxide-extraction method. 13C- and 1H-NMR spectroscopy revealed that the polyglucan consists of β-(1→3)- and β-(1→6)-linked glucose polymers, with a β-(1→6)- to β-(1→3)-linkage ratio of 1.5. Gel permeation chromatography showed that the molecular weight of the polyglucan is 1.1–8.4 × 104 Da, with a peak at 3.4 × 104 Da. The degree of polymerization, which was estimated from the amounts of total carbohydrate and reduced ends, was 203, corresponding to 3.3 × 104 Da. A method for measurement of the β-polyglucan in a small amount of liquid culture involving a mixture of β-glucanases, Westase, was established. The β-polyglucan was localized in the soluble fraction of cells. The amount of β-polyglucan per cell increased at the stationary phase under continuous illumination and decreased in the dark, like those of storage α-polyglucans, starch of green algae and glycogen of cyanobacteria. The activities of β-1,3- and β-1,6-glucanases involved in the degradation of the storage β-polyglucan were assayed in vitro, both being optimal at pH 5.0. The β-1,3-glucanase activity, which was detected on active staining after native polyacrylamide gel electrophoresis, was partially purified by ammonium sulfate precipitation and anion exchange chromatography.

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Abbreviations

DMSO:

Dimethylsulfoxide

DP:

Degree of polymerization

G6PDH:

Glucose 6-phosphate dehydrogenase

HK:

Hexokinase

TSP:

Trisodium phosphate dodecahydrate

TTC:

2,3,5-Triphenyltetrazolium chloride

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Acknowledgments

The authors thank Dr. I. Inouye of Tsukuba University, Japan, for providing the P. haptonemofera cells. They are also indebted to Mr. N. J. Halewood for his correction in English of this manuscript. This research was supported by Grant-in-Aid from the Ministry of Education, Science, Sports, and Culture, Japan, the Promotion and Mutual Aid Corporation for Private Schools of Japan.

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Authors and Affiliations

  1. School of Life Sciences, Tokyo University of Pharmacy and Life Sciences, 1432-1, Horinouchi, Hachioji, Tokyo, 192-0392, Japan

    Yasutaka Hirokawa, Shoko Fujiwara, Motoya Suzuki, Tomoka Akiyama, Minako Sakamoto, Saori Kobayashi & Mikio Tsuzuki

  2. Science and Technology, CREST, Kawaguchi, Saitama, 332-0012, Japan

    Shoko Fujiwara & Mikio Tsuzuki

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  1. Yasutaka Hirokawa
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  2. Shoko Fujiwara
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  3. Motoya Suzuki
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  6. Saori Kobayashi
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  7. Mikio Tsuzuki
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Correspondence to Shoko Fujiwara.

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Hirokawa, Y., Fujiwara, S., Suzuki, M. et al. Structural and physiological studies on the storage β-polyglucan of haptophyte Pleurochrysis haptonemofera . Planta 227, 589–599 (2008). https://doi.org/10.1007/s00425-007-0641-9

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