GROWTH AND METABOLISM OF ORAL MICROORGANISMS

口腔微生物的生长和代谢

• 批准号: 3775662
• 负责人: S A ROBRISH
• 金额: --
• 依托单位: NATIONAL INSTITUTE OF DENTAL & CRANIOFACIAL RESEARCH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3775662
• 关键词: Fusobacterium; carbohydrate metabolism; carbohydrate transport; enzyme activity; hydro lyase; maltose; mass spectrometry; microorganism culture; microorganism growth; microorganism metabolism; nuclear magnetic resonance spectroscopy; oral bacteria; phosphoenolpyruvate; phosphorylation; protein sequence

Maltose grown cells of Fusobacterium mortiferum ATCC 25557 were able to transport maltose, in contrast to cells from cultures grown on other energy sources. Maltose grown cells were also able to use both maltose and sucrose, as well as a variety of alpha linked glucosyl disaccharides, anaerobically from a buffered cell suspension. Permeabalized cells and sonic extracts of maltose grown F. mortiferum phosphorylate maltose with phosphoenolpyruvate [PEP] exclusively as the phosphate donor. Permeabalized cells from cultures grown on serine, glucose or sucrose were unable to phosphorylate maltose with either adenosine triphosphate [ATP] or PEP as a phosphate donor. PEP stimulated glucose phosphorylation was found in cells grown on all four energy sources and was strongest in maltose grown cells. When a sonic extract of F. mortiferum was resolved into membrane and cytosolic fractions neither component could affect PEP dependent maltose phosphorylation alone but maltose PTS activity was restored by addition of both components. The cystosolic fraction from either glucose or maltose grown cells complimented maltose membranes for maltose PTS activity. A glucose PTS was demonstrated in maltose grown cells. Aerobic conditions allowed PEP dependent maltose phosphorylation but prevented further disaccharide dissimilation suggesting aerobic sensitivity of a putative maltose-6-phosphate hydrolase. Maltose-6-phosphate hydrolase activity could not be demonstrated using cell extracts although evidence with intact anaerobic cells showed conclusively that the activity was present. Maltose-6-phosphate was prepared in milligram amounts using permeabalized maltose grown F. mortiferum, maltose and PEP as described herein. The structure of the compound was proved by chromatographic identification of products following enzymatic and chemical treatments. The proposed structure was confirmed using mass spectral data and nuclear magnetic resonance spectroscopy. Cystathionase [cystine lyase] was detected by activity stain in a gel following separation of sonic extracts of a variety of fusobacteria.

利用麦芽糖培养法研究了Mortiferum梭杆菌ATCC 25557的麦芽糖培养 能够运输麦芽糖，而不是培养的细胞 对其他能源的依赖。培养的麦芽糖细胞也能够利用 麦芽糖和蔗糖，以及各种阿尔法相连 来自缓冲细胞的厌氧葡萄糖二糖 停职。麦芽糖的透膜细胞和超声提取液 栽培桑白僵菌磷酸化麦芽糖与磷酸烯醇式丙酮酸 [PEP]完全作为磷酸盐供体。来自以下位置的渗透化细胞 生长在丝氨酸、葡萄糖或蔗糖上的培养物不能 用三磷酸腺苷[ATP]或 佩普作为磷酸盐捐赠者。PEP促进葡萄糖磷酸化 在所有四种能源上生长的细胞中都发现了 在麦芽糖培养的细胞中最强。当F. 桑寄生被分解为膜和胞质组分 两种成分均不影响PEP依赖的麦芽糖 单独的磷酸化，但麦芽糖的PTS活性通过 这两个组件都相加了。来自任何一种的囊泡部分 葡萄糖或麦芽糖培养的细胞称赞麦芽糖薄膜 麦芽糖PTS活性。在麦芽糖中发现了葡萄糖PTS 长成的细胞。 有氧条件允许PEP依赖的麦芽糖 磷酸化，但防止了进一步的二糖异化 推测麦芽糖-6-磷酸对氧气的敏感性 水解酶。麦芽糖-6-磷酸水解酶活性不能 使用细胞提取液演示，尽管证据完整 厌氧细胞确凿地表明活性是存在的。 以毫克量制备麦芽糖-6-磷酸 以透膜麦芽糖、麦芽糖和PEP为原料 在此描述的。化合物的结构经红外光谱确证。 酶促反应后产物的层析鉴定 化学处理。建议的结构是通过以下方式确认的 质谱学数据和核磁共振光谱学。 活性染色检测半胱氨酸酶[半胱氨酸裂解酶] 在凝胶中分离各种不同种类的 梭杆菌。