ATP的分析方法综述

本文综述了当前的ATP分析方法，比较了各方法的分析原理，优缺点及使用范围。     

蜜环菌菌丝与菌索活性成分含量研究

目的 测定蜜环菌(Armillariamellea)菌丝、菌索两种形态总多糖、总蛋白、总三萜、麦角甾酮、麦角甾醇、腺苷的含量差异。方法 采用紫外-可见分光光度法测定总三萜、总多糖及总蛋白含量。采用高效液相色谱法测定麦角甾酮、麦角甾醇、腺苷成分含量。结果 蜜环菌菌丝与菌索两种形态的大部分活性成分含量差异显著,其中总三萜、总蛋白、麦角甾醇成分显示菌丝含量优于菌索,且具有显著性差异(P<0.05);菌索总多糖含量、麦角甾酮含量优于菌丝,含量差异明显;两者腺苷含量相似。结论 对于蜜环菌的药用以及开发保健食品方面,需要根据具体方案进行合理开发,达到资源的合理利用。研究结果可为今后的综合利用开发奠定理论基础。     

Hydro‐ethanolic extract of cashew tree (Anacardium occidentale) nut and its principal compound,anacardic acid,stimulate glucose uptake in C2C12 muscle cells

Scope: Products of cashew tree (Anacardium occidentale) are used in traditional medicine for various ailments, including diabetes. Methods and results: The anti‐diabetic properties of cashew plant parts were studied using differentiated C2C12 myoblasts (myotubes) and rat liver mitochondria. Hydroethanolic extract of cashew seed (CSE) and its active component, anacardic acid (AA), stimulated glucose transport into C2C12 myotubes in a concentration‐dependent manner. Extracts of other parts (leaves, bark and apple) of cashew plant were inactive. Significant synergistic effect on glucose uptake with insulin was noticed at 100 μg/mL CSE. CSE and AA caused activation of adenosine monophosphate‐activated protein kinase in C2C12 myotubes after 6 h of incubation. No significant effect was noticed on Akt and insulin receptor phosphorylation. Both CSE and AA exerted significant uncoupling of succinate‐stimulated respiration in rat liver mitochondria. Conclusion: Activation of adenosine monophosphate‐activated protein kinase by CSE and AA likely increases plasma membrane glucose transporters, resulting in elevated glucose uptake. In addition, the dysfunction of mitochondrial oxidative phosphorylation may enhance glycolysis and contribute to increased glucose uptake. These results collectively suggest that CSE may be a potential anti‐diabetic nutraceutical.     

Optimisation and significance of ATP analysis for measuring active biomass in granular activated carbon filters used in water treatment

A method for determining the concentration of active microbial biomass in granular activated carbon (GAC) filters used in water treatment was developed to facilitate studies on the interactions between adsorption processes and biological activity in such filters. High-energy sonication at a power input of 40 W was applied to GAC samples for the detachment of biomass which was measured as adenosine triphosphate (ATP). Modelling of biomass removal indicated that a series of six to eight sonication treatments of 2 min each yielded more than 90% of the attached active biomass. The ATP concentrations in 30 different GAC filters at nine treatment plants in The Netherlands ranged from 25 to 5000 ng ATP cm(-3) GAC, with the highest concentrations at long filter run times and pretreatment with ozone. A similar concentration range was observed in nine rapid sand (RS) filters. ATP concentrations correlated significantly (p<0.05) with total direct bacterial cell counts in each of these filter types, but the median value of the ATP content per cell in GAC filters (2.1 x 10(-8) ng ATP/cell) was much lower than in the RS filters (3.6 x 10(-7) ng ATP/cell). Average biofilm concentrations ranging from 500 to 10(5) pg ATP cm(-2) were calculated assuming spherical shapes for the GAC particles but values were about 20 times lower when the surface of pores >1 microm diameter is included in these calculations. The quantitative biomass analysis with ATP enables direct comparisons with biofilm concentrations reported for spiral wound membranes used in water treatment, for distribution system pipes and other aquatic environments.     

Effect of gamma-irradiation on flavour 5′-nucleotides,tyrosine, and phenylalanine in mushrooms (Agaricus bisporus)

The impact of gamma-irradiation on 5′-nucleotides and on the free amino acids tyrosine and phenylalanine in fresh mushrooms was studied. After irradiation the samples were freeze-dried to avoid enzyme induced chemical changes. Three 5′-nucleotides could be detected using HPLC–UV and LC–ESI-MS: adenosine 5′-monophosphate (AMP), guanosine 5′-monophosphate (GMP) and guanosine 5′-diphosphate (GDP). Irradiation significantly reduced (p = 0.05) the GDP concentration (22%). AMP showed a marked reduction (46%) only at 5 kGy. GMP, tyrosine, and phenylalanine were not affected by gamma-irradiation.     

Biomass Recovery Method for Adenosine Triphosphate (ATP) Quantification Following UV Disinfection

A biomass recovery method was developed to monitor UV disinfection efficacy using adenosine triphosphate (ATP). Typically, disinfection monitoring at wastewater treatment facilities (WWTFs) involves quantifying fecal and total coliforms or colony forming units, the results of which take a minimum of 24 h to produce. ATP quantification immediately before and after UV treatment, which takes only minutes, shows little reduction and often an increase in the microbial population since UV irradiation results in cells that are viable (i.e., still producing ATP) but not culturable. To overcome this, our biomass recovery method incorporates an incubation step to encourage life cycling of microbes. Average log reductions in cellular ATP (cATP) were found to be ?0.28 ± 0.19, ?0.011 ± 0.153, ?0.17 ± 0.32, and 0.065 ± 0.074 using direct ATP measurements on UV-treated samples from WWTFs A, B, C, and D, respectively, while those using the recovery method were correspondingly 0.17 ± 0.34, 1.8 ± 0.8, 0.20 ± 0.35, and 0.72 ± 0.26. The response of the biomass recovery-ATP method indicated a significant direct correlation to the microbial population reduction observed in heterotrophic plate count (HPC) and Colilert® methods using both pure Escherichia coli culture and secondary municipal wastewater effluent.     

Enzyme-assisted extraction of cordycepin and adenosine from cultured Cordyceps militaris and purification by macroporous resin column chromatography

Enzyme-assisted extraction (EAE) coupled with macroporous resin column chromatography (MRCC) was successfully used for the extraction and purification of cordycepin and adenosine from cultured Cordyceps militaris. After optimized by the response surface methodology (RSM), the extraction yields of cordycepin and adenosine were 86.45% and 70.06%, respectively. Subsequently, under the optimal separation conditions on NKA-II resins (loading 1 BV of extraction solution with pH 8.0, and eluting with 4 BVs of 70% ethanol at a flow rate of 3 BV/h), cordycepin (purity of 32.5%), and adenosine (purity of 39.9%) were obtained with an overall recovery rate more than 90%.     

生物发光法细菌快速检测仪的研制及应用

基于ATP生物发光法,研制了一种细菌快速检测仪。该检测仪内置高精度A/D转换器,以小型光电倍增管进行光电转换,结合实验室自制的生物传感器,集光路、电路及软件设计于一体,组成了完整的细菌快速检测系统。其用于大肠杆菌标准品溶液检测时,检测结果与培养计数法结果的相关系数(R)达到0.9760,同时获得了仪表检测光强和细菌浓度关系曲线。针对实际样品检测中存在的问题,提出了相应的数据处理改进算法,引入了校准方程修正系数,用于食品及卡介苗样品检测,获得了良好的检测结果。对猴头菇类食品样品检测的结果与培养计数法检测结果的R达到0.993,重复性检测的相对平均偏差(R.A.D.)为7.69%,变异系数(CV)为9.07%。对卡介苗(BCG)样品检测结果与培养计数法检测结果的R达到0.997,R.A.D.为5.72%,CV为7.73%。与传统培养计数法相比,该仪表具有较好的检测速度,准确度和重复性。     

Antimicrobial effects of cinnamon bark oil on microbial composition and quality of grass carp (Ctenopharyngodon idellus) fillets during chilled storage

In this study, 11 essential oils were initially screened for antimicrobial activity against Aeromonas veronii, Acinetobacter johnsonii, Shewanella putrefaciens, and Pseudomonas jessenii. Cinnamon bark oil exhibited the highest antimicrobial activity and was used as a natural preservative in grass carp fillets stored at 4 ± 1 °C. Sensory assessment, total volatile basic nitrogen (TVB-N), color, biogenic amines, adenosine triphosphate (ATP)-related compounds, K-value, and presence of microbiota were analyzed during storage. Cinnamon bark oil treatment was found to be effective in enhancing organoleptic quality, inhibiting microbial growth, and delaying the increase of TVB-N, putrescine, cadaverine and K-value. Based on sensory analysis, shelf-life of grass carp fillets was 8 days for the control and 12 days for the treatment group. According to high-throughput sequencing, Aeromonas and Pseudomonas were the predominant microbiota in fresh grass carp. As storage time progressed, the microbial composition of both control and treatment samples became less diverse. Pseudomonas followed by Aeromonas were dominant in spoiled control samples. However, there were significant differences in microbial composition between control and treatment samples at the end of storage. Cinnamon bark oil treatment inhibited the growth of Aeromonas and Shewanella, and Pseudomonas was the only predominant microbiota found in spoiled treatment samples.