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蜜蜂葡萄糖氧化酶——其在工蜂中的表达以及天然蜂蜜中其含量与过氧化氢介导的抗菌活性的比较分析

Honeybee glucose oxidase--its expression in honeybee workers and comparative analyses of its content and H2O2-mediated antibacterial activity in natural honeys.

作者信息

Bucekova Marcela, Valachova Ivana, Kohutova Lenka, Prochazka Emanuel, Klaudiny Jaroslav, Majtan Juraj

机构信息

Faculty of Natural Sciences, Comenius University, Mlynska Dolina, 842 15, Bratislava, Slovakia.

出版信息

Naturwissenschaften. 2014 Aug;101(8):661-70. doi: 10.1007/s00114-014-1205-z. Epub 2014 Jun 27.

Abstract

Antibacterial properties of honey largely depend on the accumulation of hydrogen peroxide (H2O2), which is generated by glucose oxidase (GOX)-mediated conversion of glucose in diluted honey. However, honeys exhibit considerable variation in their antibacterial activity. Therefore, the aim of the study was to identify the mechanism behind the variation in this activity and in the H2O2 content in honeys associated with the role of GOX in this process. Immunoblots and in situ hybridization analyses demonstrated that gox is solely expressed in the hypopharyngeal glands of worker bees performing various tasks and not in other glands or tissues. Real-time PCR with reference genes selected for worker heads shows that the gox expression progressively increases with ageing of the youngest bees and nurses and reached the highest values in processor bees. Immunoblot analysis of honey samples revealed that GOX is a regular honey component but its content significantly varied among honeys. Neither botanical source nor geographical origin of honeys affected the level of GOX suggesting that some other factors such as honeybee nutrition and/or genetic/epigenetic factors may take part in the observed variation. A strong correlation was found between the content of GOX and the level of generated H2O2 in honeys except honeydew honeys. Total antibacterial activity of most honey samples against Pseudomonas aeruginosa isolate significantly correlated with the H2O2 content. These results demonstrate that the level of GOX can significantly affect the total antibacterial activity of honey. They also support an idea that breeding of novel honeybee lines expressing higher amounts of GOX could help to increase the antibacterial efficacy of the hypopharyngeal gland secretion that could have positive influence on a resistance of colonies against bacterial pathogens.

摘要

蜂蜜的抗菌特性很大程度上取决于过氧化氢(H2O2)的积累,过氧化氢是由葡萄糖氧化酶(GOX)介导稀释蜂蜜中的葡萄糖转化而产生的。然而,不同蜂蜜的抗菌活性存在显著差异。因此,本研究的目的是确定这种活性差异以及蜂蜜中H2O2含量差异背后的机制,这与GOX在此过程中的作用有关。免疫印迹和原位杂交分析表明,gox仅在执行各种任务的工蜂下咽腺中表达,而在其他腺体或组织中不表达。对工蜂头部选择的参考基因进行实时PCR分析表明,gox的表达随着最年轻的蜜蜂和哺育蜂的衰老而逐渐增加,并在采集蜂中达到最高值。对蜂蜜样本的免疫印迹分析表明,GOX是蜂蜜中的一种常见成分,但其含量在不同蜂蜜之间存在显著差异。蜂蜜的植物来源和地理来源均不影响GOX的水平,这表明一些其他因素,如蜜蜂营养和/或遗传/表观遗传因素,可能参与了观察到的差异。除了甘露蜜外,蜂蜜中GOX的含量与产生的H2O2水平之间存在很强的相关性。大多数蜂蜜样本对铜绿假单胞菌分离株的总抗菌活性与H2O2含量显著相关。这些结果表明,GOX的水平可以显著影响蜂蜜的总抗菌活性。它们还支持这样一种观点,即培育表达更高量GOX的新型蜜蜂品系可能有助于提高下咽腺分泌物的抗菌功效,这可能对蜂群抵抗细菌病原体产生积极影响。

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