Alhudhud M, Humphreys P, Laws A
Department of Chemical and Biological Sciences, University of Huddersfield, Queensgate, Huddersfield HD1 3DH, UK.
J Microbiol Methods. 2014 May;100:93-8. doi: 10.1016/j.mimet.2014.02.021. Epub 2014 Mar 13.
Exopolysaccharides (EPSs) produced by Bifidobacteria have received considerable attention due to their ability to modify the rheological and physicochemical properties of dairy products. However, the quantification and characterisation of Bifidobacterial EPS are hampered by the presence of EPS-equivalent (EPS-E) substances in complex media such as Reinforced Clostridial Medium (RCM). This study has developed a medium based on RCM which both supports the growth of Bifidobacterium animalis ssp. lactis AD011 and does not interfere with the quantification and characterisation of the EPS generated. Medium development involved the identification of EPE-E containing components via NMR analysis followed by their removal, substitution or pre-treatment. Both beef extract and casein acid hydrolysate required chemical pre-treatment to remove polysaccharide components before the medium was free of EPS-E materials. Once EPS-E free components had been identified, lactose, glucose and galactose were evaluated as potential carbon sources. Glucose was found to be the optimum carbon source. The final medium composition supported growth to the same extent as RCM providing significant EPS yields and no interferences during analysis.
由于双歧杆菌产生的胞外多糖(EPSs)能够改变乳制品的流变学和物理化学性质,因此受到了广泛关注。然而,在诸如强化梭菌培养基(RCM)等复杂培养基中存在EPS等效物(EPS-E)物质,这阻碍了双歧杆菌EPS的定量和表征。本研究开发了一种基于RCM的培养基,该培养基既能支持动物双歧杆菌亚种乳酸双歧杆菌AD011的生长,又不会干扰所产生EPS的定量和表征。培养基的开发包括通过NMR分析鉴定含EPE-E的成分,然后将其去除、替换或预处理。牛肉提取物和酪蛋白酸水解物在培养基不含EPS-E物质之前都需要进行化学预处理以去除多糖成分。一旦确定了不含EPS-E的成分,就对乳糖、葡萄糖和半乳糖作为潜在碳源进行了评估。发现葡萄糖是最佳碳源。最终培养基组成支持的生长程度与RCM相同,提供了显著的EPS产量,并且在分析过程中没有干扰。