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碳源和氮源如何影响β-(1,3/1,6)-葡聚糖的合成、其结构以及产朊假丝酵母细胞对β-(1,3)-葡聚糖单克隆抗体免疫标记的敏感性?

How do the carbon and nitrogen sources affect the synthesis of β-(1,3/1,6)-glucan, its structure and the susceptibility of Candida utilis yeast cells to immunolabelling with β-(1,3)-glucan monoclonal antibodies?

机构信息

Department of Food Biotechnology and Microbiology, Institute of Food Sciences, Warsaw University of Life Sciences, Nowoursynowska 159C Street, 02-787, Warsaw, Poland.

Department of Glycobiotechnology, Institute of Chemistry Slovak Academy of Sciences, Dúbravská Cesta 9, 84538, Bratislava, Slovakia.

出版信息

Microb Cell Fact. 2024 Jan 19;23(1):28. doi: 10.1186/s12934-024-02305-4.

Abstract

BACKGROUND

The 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.

RESULTS

The 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.

CONCLUSIONS

Obtained 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.

摘要

背景

由于致病微生物对这些药物的抗药性不断扩散,需要限制抗生素治疗,这促使人们研究新的治疗药物,包括动物疾病的治疗和预防。这是欧洲绿色协议和从农场到餐桌战略的目标之一。具有适当组成和细胞壁多糖暴露的酵母生物质可以在精准动物营养中构成功能性饲料添加剂,自然刺激免疫系统抵抗感染。

结果

本研究进行的研究结果表明,根据生长培养基的不同,产朊假丝酵母 ATCC 9950 酵母生物质的组成也有所不同,特别是β-(1,3/1,6)-葡聚糖、α-葡聚糖和海藻糖的含量。在以去蛋白土豆汁(DPJW)为氮源和甘油为碳源的条件下培养时,观察到β-(1,3/1,6)-葡聚糖的含量最高。从酵母生物质中分离出的多糖证实,在上述培养基中培养后β-(1,3/1,6)-葡聚糖的产量最高。根据培养条件的不同,细胞内定位的β-(1,3)-葡聚糖与特异性β-(1,3)-葡聚糖抗体相互作用的敏感性也不同。有趣的是,在用甘油作为碳源和酵母提取物加蛋白胨作为氮源的培养基中细胞增殖后,添加甘油和半乳糖的 DPJW 中细胞内的聚合物与单克隆抗体的结合强度最高,而对这种相互作用的敏感性较低。

结论

获得的结果证实,考虑到侧链长度和支化频率,β-(1,3/1,6)-葡聚糖聚合物的结构存在差异,以及β-(1,3)-和β-(1,6)-链的数量存在差异,但在分离聚合物的结构特征与其在整个细胞中免疫标记的敏感性之间没有观察到明显的关系。推测,其他外表面成分和分子可以掩盖、屏蔽、保护或隐藏抗体的表位。β-(1,3)-葡聚糖在含有较低海藻糖和糖原的细胞中被单克隆抗体更强烈地识别。这表明需要在适当的条件下培养酵母生物质,以发挥可能的治疗功能。然而,我们的体外发现还需要在使用组织或动物模型的进一步研究中得到证实。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3bb1/10799355/9531b7e58579/12934_2024_2305_Fig1_HTML.jpg

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