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Kre6 (yeast 1,6-β-transglycosylase) homolog, PhTGS, is essential for β-glucan synthesis in the haptophyte Pleurochrysis haptonemofera.

ABSTRACT: Haptophytes synthesize unique β-glucans containing more β-1,6-linkages than β-1,3 linkages, as a storage polysaccharide. To understand the mechanism of the synthesis, we investigated the roles of Kre6 (yeast 1,6-β-transglycosylase) homologs, PhTGS, in the haptophyte Pleurochrysis haptonemofera. RNAi of PhTGS repressed β-glucan accumulation and simultaneously induced lipid production, suggesting that PhTGS is involved in β-glucan synthesis and that the knockdown leads to the alteration of the carbon metabolic flow. PhTGS was expressed more in light, where β-glucan was actively produced by photosynthesis, than in the dark. The crude extract of E. coli expressing PhKre6 demonstrated its activity to incorporate ¹⁴C-UDP-glucose into β-glucan of P. haptonemofera. These findings suggest that PhTGS functions in storage β-glucan synthesis specifically in light, probably by producing the β-1,6-branch.

SUBMITTER: Inukai M

PROVIDER: S-EPMC10543733 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature

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Kre6 (yeast 1,6-β-transglycosylase) homolog, PhTGS, is essential for β-glucan synthesis in the haptophyte Pleurochrysis haptonemofera.

Inukai Mayuka M Kobayashi Naoya N Endo Hirotoshi H Asakawa Koki K Amano Keisuke K Yasuda Yuki Y Cenci Ugo U Colleoni Christophe C Ball Steven S Fujiwara Shoko S

Frontiers in bioengineering and biotechnology 20230918

Haptophytes synthesize unique β-glucans containing more β-1,6-linkages than β-1,3 linkages, as a storage polysaccharide. To understand the mechanism of the synthesis, we investigated the roles of Kre6 (yeast 1,6-β-transglycosylase) homologs, PhTGS, in the haptophyte Pleurochrysis haptonemofera. RNAi of PhTGS repressed β-glucan accumulation and simultaneously induced lipid production, suggesting that PhTGS is involved in β-glucan synthesis and that the knockdown leads to the alterat ...[more]

PMID: 37790259

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Project description:BackgroundThe need to limit antibiotic therapy due to the spreading resistance of pathogenic microorganisms to these medicinal substances stimulates research on new therapeutic agents, including the treatment and prevention of animal diseases. This is one of the goals of the European Green Deal and the Farm-To-Fork strategy. Yeast biomass with an appropriate composition and exposure of cell wall polysaccharides could constitute a functional feed additive in precision animal nutrition, naturally stimulating the immune system to fight infections.ResultsThe results of the research carried out in this study showed that the composition of Candida utilis ATCC 9950 yeast biomass differed depending on growth medium, considering especially the content of β-(1,3/1,6)-glucan, α-glucan, and trehalose. The highest β-(1,3/1,6)-glucan content was observed after cultivation in deproteinated potato juice water (DPJW) as a nitrogen source and glycerol as a carbon source. Isolation of the polysaccharide from yeast biomass confirmed the highest yield of β-(1,3/1,6)-glucan after cultivation in indicated medium. The differences in the susceptibility of β-(1,3)-glucan localized in cells to interaction with specific β-(1,3)-glucan antibody was noted depending on the culture conditions. The polymer in cells from the DPJW supplemented with glycerol and galactose were labelled with monoclonal antibodies with highest intensity, interestingly being less susceptible to such an interaction after cell multiplication in medium with glycerol as carbon source and yeast extract plus peptone as a nitrogen source.ConclusionsObtained results confirmed differences in the structure of the β-(1,3/1,6)-glucan polymers considering side-chain length and branching frequency, as well as in quantity of β-(1,3)- and β-(1,6)-chains, however, no visible relationship was observed between the structural characteristics of the isolated polymers and its susceptibility to immunolabeling in whole cells. Presumably, other outer surface components and molecules can mask, shield, protect, or hide epitopes from antibodies. β-(1,3)-Glucan was more intensely recognized by monoclonal antibody in cells with lower trehalose and glycogen content. This suggests the need to cultivate yeast biomass under appropriate conditions to fulfil possible therapeutic functions. However, our in vitro findings should be confirmed in further studies using tissue or animal models.

Dataset Information

Kre6 (yeast 1,6-β-transglycosylase) homolog, PhTGS, is essential for β-glucan synthesis in the haptophyte Pleurochrysis haptonemofera.

Publications

Kre6 (yeast 1,6-β-transglycosylase) homolog, PhTGS, is essential for β-glucan synthesis in the haptophyte <i>Pleurochrysis haptonemofera</i>.

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