Kinetic Model for Invertase‐Induced Sucrose Hydrolysis: Initial Time Lag

The kinetics of sucrose hydrolysis by invertase was studied in order to find a comprehensive model for the reaction pathway and mechanism. First, three common models of Michaelis‐Menten (MM), substrate inhibition (S²), and substrate clusters' inhibition (S³(I)) were investigated. The third model was found to better predict the initial sucrose concentration. Then, the S³(I) model was modified to cover the remaining pathway (S³(II)). Finally, a new comprehensive model (S³(III)) was evaluated, which in addition to what is considered in the two previously mentioned models (S³(I) and S³(II)) also involved the initial time lag. The model predictions showed an excellent agreement with the experimental data. The mean absolute error for the MM model is significantly reduced for the S³(III) model.     

Residual effect of atomised water vapour treatment on carbohydrate metabolism during ripening of cv “Fino de Jete” cherimoya fruit

Atomised hot water vapour at between 46 and 54 °C for 60 min was tested as a method of delaying ripening and extending shelf life in cv “Fino de Jete” cherimoya fruit (Annona cherimola Mill.). Heat treatment at 46 °C stimulated the respiration rate and starch degradation, induced ethylene production, increased the accumulation of soluble sugars and α-amylase activity, and moved the onset of ethylene production (OEP) forward. Treatment at 50 °C, and especially at 52 °C, delayed the climacteric peak and the OEP, decreased sugar and organic acid accumulation, and reduced α-amylase and invertase activity. The results obtained indicate that temperatures higher than 48 °C slowed physiological processes and carbohydrate metabolism and extend the shelf life of the cherimoya fruits, while temperatures higher than 52 °C blocked ripening irreversibly.     

Novel sucrose three-enzyme conductometric biosensor

For the first time a conductometric biosensor for sucrose determination has been developed using a complex three-enzyme (invertase, mutarotase, and glucose oxidase) containing membrane as a sensitive element immobilized on the conductometric interdigitated planar electrodes. The time of measurement of sucrose concentration in the solution was about 1–2 min. The dynamic range of biosensor depends on buffer capacity, being 2 μM–5 mM of sucrose in 5 mM phosphate buffer. The conductometric biosensor developed demonstrates high selectivity, operational stability and reproducibility. The dependence of sensor response on pH and ionic strength of tested solution has been studied in this work. Storage conditions have also been under investigation. The sensor appeared to be eligible towards application in practice.     

Enzyme activity and biochemical changes of three date palm cultivars with different softening pattern during ripening

The physicochemical composition changes during palm ripening were studied. The activities of invertase and cell wall-modifying enzymes, namely pectin methylesterase (PME), β-galactosidase (β-Gal), endo-1,4-β-d-glucanase (EGase), and cellulase were monitored during growth and ripening of Shahani, Piarom and Deiry cultivars with different textures. Also, we estimated the concentrations of six organic acids by HPLC. Reducing sugars, most of organic acid, pH and TSS increased up to the full-ripe stage of all date types. On the contrary, moisture, macro and microelements and proteins decreased during the same period. Despite a considerable rise in invertase activity (200 units) during ripening of Shahani, no significant trend could be discerned in Deiry cultivar (8 units) at different stages. Our results also showed that cell wall enzymes were increased in activity during ripening and these increases were coincident with fruit softening. Furthermore, our results showed that the composition and the variation of the chemical compositions mainly depended on the cultivar and maturity stage.     

Polyacrylamide–gelatine carrier system used for invertase immobilization

Invertase was immobilized into polyacrylamide–gelatin carrier system by chemical cross-linking with chromium (III) acetate, chromium (III) sulphate, and potassium chromium (III) sulphate. The optimum conditions, namely substrate concentration, temperature, and pH were determined. The effect of polyacrylamide–gelatin ratio and cross-linker concentration on immobilized enzyme activity were analysed. Maximum immobilized enzyme activities were obtained with chromium (III) acetate (0.01 mol dm⁻³), chromium (III) sulphate (0.004 mol dm⁻³) and potassium chromium (III) sulphate (0.001 mol dm⁻³) for 0.177 (w/w) polyacrylamide–gelatin carrier ratio as 79%, 72% and 79%, respectively. The K_m values were 86 and 166 mM for free and immobilized enzyme, respectively. All immobilized samples were used 20 times over a period of 2 months without a considerable loss of activity.     

从啤酒酿造酵母中提取蔗糖酶及酶的性质研究

采用SDS抽提法从啤酒酵母中提取蔗糖酶,通过正交实验对SDS浓度、提取时间、pH、提取温度等条件进行优化,确定了最佳提取工艺条件是：SDS浓度0.6mmol/L,温度50℃,时间10h,pH5.0。制得的粗酶经醇沉后,采用Q-琼脂糖凝胶FF阴离子交换层析方法得到纯化的酵母蔗糖酶,纯化倍数为16.8倍,并对蔗糖酶的酶性质进行了研究。     

Glass transition and water effects on sucrose inversion in noncrystalline carbohydrate food systems

The effects of water and glass transition on the hydrolysis of sucrose by invertase in noncrystalline carbohydrate systems were investigated. Maltodextrin/sucrose (2:1) and maltodextrin/lactose/sucrose (1:1:1) were dissolved in distilled water. Invertase (10 mg/17.2 g) was added. Amorphous samples were produced by freeze-drying the solutions. Sorption isotherms were determined gravimetrically at 24 °C over the 0.113–0.763 a_w, and over 0.239–0.764 a_w, the glass transition, T_g was determined using differential scanning calorimetry (DSC). DSC and water sorption results suggested that samples remained noncrystalline. Sucrose inversion was analysed by monitoring glucose content during storage. Sucrose hydrolysis occurred at significant rates at 0.662 and 0.764 a_w. The rate increase was not related to the apparent glass transition of the systems.     

不同氮量与氮源下烤烟淀粉酶和转化酶活性动态变化

研究结果表明,在烟叶生长成熟过程中,烟叶中淀粉酶活性变化呈双峰曲线,峰值分别出现在叶片功能盛期和成熟期。在烟叶成熟期以前淀粉酶活性与施氮量呈正相关,成熟期则呈负相关。转化酶活性在叶片功能盛期后逐渐降低,随着施氮水平的提高,转化酶活性提高。     

Enzymatic activities related to cherimoya fruit softening and sugar metabolism during short-term controlled-atmosphere treatments

 The effects of short controlled-atmosphere (CA) treatments, involving exposure to 10% O₂ combined with 10%, 15% and 20% CO₂, on a selection of biochemical parameters and related enzymatic activities of cherimoya fruit were studied. A marked increase in the activity of soluble and cell-wall invertases was found in cherimoyas stored in air, but this was inhibited by CA treatment, which also reduced the accumulation of soluble sugar and malic acid. CA had no effect on the citric acid content and enhanced the rise in fumaric acid concentration, which was directly related to the CO₂ level. Differences in the softening of fruit stored in air and in CA were found after 9 days of storage, but the role of the cell-wall hydrolases polygalacturonase and carboxymethyl-cellulase in this behaviour was not elucidated. Received: 25 January 1998 / Revised version: 3 March 1998     

便携式血糖仪对耐甲氧西林金黄色葡萄球菌mecA基因的灵敏检测方法研究

本实验利用血糖仪构建了一种基于蔗糖酶修饰的纳米金功能化树状大分子对耐甲氧西林金黄色葡萄球菌mecA基因的灵敏检测新方法。当靶基因存在时,其与捕获核酸探针特异性结合,释放出带蔗糖酶的信号分子复合物,经磁分离后,上清液中的蔗糖酶催化底物蔗糖,产生可检测的葡萄糖信号,从而实现血糖仪对mecA基因的定量检测。该检测方法的线性范围是0.5pM到1nM,检测限为0.26pM,优于一般的基因检测方法。结果表明,该方法在疾病诊断方面具有潜在的应用价值。