DNA-based carrier detection and prenatal diagnosis of tyrosinase-negative oculocutaneous albinism (OCA1A)

We describe molecular prenatal diagnosis and carrier detection of tyrosinase-negative oculocutaneous albinism (OCA1A) in two families. In one family, we carried out DNA-based prenatal diagnosis of OCA1A. In the other family, mutation analysis and carrier detection obviated the need for prenatal diagnosis. Molecular analysis is safer and probably more accurate than fetoscopy and fetal scalp biopsy, and should become the method of first choice for prenatal diagnosis of OCA1.     

磁场对酪氨酸酶活性及其催化降解酚类有机物影响的研究

研究了恒磁场对酪氨酸酶(TYR)活性及其催化降解酚类有机物的影响.结果显示,不同磁场强度(10～350 mT)处理下TYR酶活性均有提高,最佳磁场强度150mT下酶活力提高了27.1%.不同磁化时间下TYR酶活都有上升,但磁化60min后酶活上升幅度有所下降.酶经磁场处理后对温度、pH值稳定性增强,在温度为20～35 ℃、pH为5.0～10.0时均能保持较高活性,最佳温度为25℃,最佳pH为7.0;磁化后TYR酶的Michaelis常数Km为3.83 mmol·L-1,未磁化的Michaelis常数Km为2.65 mmol·L-1.磁场作用可促进TYR对酚类有机物邻苯二酚、苯酚、2,4-二氯酚的转化,反应速度依次递减,磁化处理对邻苯二酚反应的促进作用尤其明显;磁化处理后的酶对不同浓度苯酚和2,4-二氯酚的去除率均明显高于未磁化处理的酶,且随着酶用量增加,酚去除率提高.荧光发射光谱分析表明,磁化酪氨酸酶的荧光强度增强,构像发生了变化.     

Renewable Resources and Enzymatic Processes to Create Functional Polymers: Adapting Materials and Reactions from Food Processing

We are exploiting materials and concepts from food science to create functionalized, environmentally friendly derivatives of the biopolymer chitosan, a byproduct of seafood processing. Functional groups are grafted onto chitosan using tyrosinase, the enzyme responsible for food browning. The functionalizing groups studied include low-molecular-weight phenols derived from natural sources and high-molecular-weight proteins. The approach of using low-molecular-weight phenols to functionalize chitosan is illustrated with arbutin, a natural phenol found in pears. Results demonstrate that tyrosinase initiates reactions that lead to the conversion of arbutin–chitosan solutions into gels. These gels can be rapidly broken by treatment with the chitosan-hydrolyzing enzyme chitosanase, demonstrating that the chitosan derivatives remain biodegradable. We briefly review other studies in which low-molecular-weight natural phenols are enzymatically grafted onto chitosan to confer functional properties. The creation of co-polymers is illustrated by results in which tyrosinase is used to couple gelatin onto chitosan. Gelatin is a proteinaceous byproduct of meat production. The tyrosinase-generated gelatin–chitosan conjugates have been observed to offer interesting rheological and thermal properties. These results demonstrate the potential for using renewable resources and enzymatic processing to create environmentally friendly polymers with useful functional properties.     

苯酚在预活化聚酰胺膜酪氨酸酶生物传感器上的响应特性

研究苯酚在预活化聚酰胺膜酪氨酸酶生物传感器上的响应特性,适量儿茶酚的存在可加快传感器到达恒态的速度。被固定酶的量、活性及传感器的有效面积等是影响检测灵敏度的主要因素。首次采用预活化聚酰胺膜固定酪氨酸酶,并与浸蜡石墨电极组合成酪氨酸酶生物传感器,在温度２５℃电位－０．２００ＶｖｓＳＣＥ,儿茶酚浓度５×１０（－７）ｍｏｌ／Ｌ,底液０．１ｍｏｌ／Ｌ磷酸盐缓冲溶液ｐＨ６．５０下,采用电流法测定苯酚及水样中的酚。酶膜的制备、替换和贮存极为方便,酶膜贮存５个月未见活性下降,有良好的重现性和线性范围２×１０（－７）－１．２５×１０（－５）ｍｏｌ／Ｌ。     

Screening the methanolic extracts of some plants for tyrosinase inhibitory activity

Melanin is responsible for pigmentation of skin and hair. It protects the skin from the harmful effects of UV radiation; however, its abnormal production may lead to different diseases such as hyper-pigmentation or vitiligo. Tyrosinase is a key enzyme in melanin production, therefore inhibitors of this enzyme should have broad application. Due to the adverse effects of synthetic inhibitors currently being used, the search for new inhibitors, especially natural ones, is needed. The methanolic extracts of 80 plants were tested for their tyrosinase inhibitory activities by spectrophotometry at 492?nm. The inhibition mode of effective plant extracts was determined using the Lineweaver-Burk plots. Among these plant extracts, Alhagi camelorum (camel thorn), Camelia sinensis (green tea), Cuminum cyminum (Cumin), Lawsonia inermis (henna), Quercus infectoria (masso), and Vaccinium arctostaphylos (Caucasian whortleberry) were identified as tyrosinase inhibitors with inhibitory effect of more than 50% at concentrations of 1.14?g?L^?1. The inhibition mode of four newly found effective plant extracts showed that A. camelorum and V. arctostaphylos inhibit the enzyme in a competitive manner whereas L. inermis and Q. infectoria have mixed inhibitory effect. To recommend these plant extracts for hyper-pigmentation disorders and other applications, further in vitro and in vivo studies are needed.     

麦芽糊精修饰测酚酪氨酸酶膜电极的研制

研究了用麦芽糊精修饰的电流型酪氨酸酶膜电极测水中酚类污染物质的方法。电极的工作条件为 :工作电位- 1 0 0mV ,工作pH值 5 4 0 ,测量时间 3min。在此测量条件下 ,所研制电极对苯酚浓度的检测下限为 1 0 0×1 0 - 6 mol L ,线性区间为 1 0 0× 1 0 - 6 ～ 1 0 0× 1 0 - 4mol L ,精密度和准确度良好 ,分别为 2 5 %和 0 2 0 % ,并具有一定的抗阴阳离子干扰的能力。此电极对其它酚类物质如邻苯二酚、对氯苯酚、间甲酚等都有良好的响应。在使用 72h后 ,电极响应最好 ,1周后电极仍可用于检测     

Tyrosinase-modified carbon felt-based flow-biosensors: The role of ultra-sonication in shortening the enzyme immobilization time and improving the sensitivity for p-chlorophenol

Tyrosinase (TYR) was covalently immobilized onto amino-functionalized carbon felt surface via glutaraldehyde-coupling under ultrasonic treatment for 10 min. The resulting TYR-immobilized carbon felt was used as a working electrode unit of bioelectrocatalytic flow-through detector for TYR substrates (catechol, p-chlorophenol (p-CP), p-cresol, phenol etc.). Cathodic peak currents based on the electroreduction of enzymatically produced o-quinones were detected at –50 mV vs. Ag/AgCl. Compared with previous work in which TYR was immobilized onto amino-functionalized carbon felt for 16 hr without the ultrasonic treatment, we succeeded in (1) shortening the enzyme immobilization time from 16 hr to 10 min, (2) enhancing the sensitivity of p-CP, and (3) improving the operational stability of p-CP. The ultrasonic treatment during the TYR immobilization step would lead to certain changes in the structure of the immobilized TYR and the morphology of the immobilized TYR-layer on the carbon felt surface.     

Rapid method for on-site determination of phenolic contaminants in water using a disposable biosensor

A disposable biosensor was fabricated using single-walled carbon nanotubes, gold nanoparticles and tyrosinase (SWCNTs-AuNPs-Tyr) modified screen-printed electrodes. The prepared biosensor was applied to the rapid determination of phenolic contaminants within 15 minutes. The SWCNTs-AuNPs-Tyr bionanocomposite sensing layer was characterized with scanning electron microscopy, electrochemical impedance spectroscopy and cyclic voltammetry methods. The characterization results revealed that SWCNTs could lead to a high loading of tyrosinase (Tyr) with the large surface area and the porous morphology, while AuNPs could retain the bioactivity of Tyr and enhance the sensitivity. The detection conditions, including working potential, pH of supporting electrolyte and the amount of Tyr were optimumed. As an example, the biosensor for catechol determination displayed a linear range of 8.0 × 10^-8 to 2.0 × 10^-5 mol·L^-1 with a detection limit of 4.5 × 10^-8 mol·L^-1 (S/N = 3). This method has a rapid response time within 10 s, and shows excellent repeatability and stability. Moreover, the resulting biosensor could be disposable, low-cost, reliable and easy to carry. This kind of new Tyr biosensor provides great potential for rapid, on-site and cost-effective analysis of phenolic contaminants in environmental water samples.