Development of a method to quantify sucrose in soybean grains

The sucrose content of soybean seeds affects the final flavor of soy-derived products. The aim of this work was to develop a simple, low-cost, spectrophotometric method for sucrose quantification in soybean seeds. To achieve this goal, we combined the action of invertase, an enzyme that hydrolyses sucrose into fructose and glucose, with glucose oxidase, an enzyme widely used for glucose quantification. This system was adapted to ELISA plates, making large-scale analyses possible at low cost, with potential application in routine analyses. To validate this method, sucrose content was determined in seeds of 14 soybean cultivars by this new method, as well as by HPLC and the enzymatic method of Stitt. The correlation coefficients were high and significant between the results obtained with the new method and the HPLC method (r = 0.9766) and the Stiff method (0.9461).     

Sugar and acid-related quality attributes and enzyme activities in strawberry fruits: Invertase is the main sucrose hydrolysing enzyme

This paper presents sugar and organic acid content and composition, the principal determinants of flavour, of strawberries of the cultivars Festival and Ventana at four consecutive developmental stages. The metabolite profiles of these two cultivars were very similar. Glucose was the predominant sugar at approximately twice the concentration of fructose and sucrose. Total acid concentrations were five times lower than total sugar concentrations in ripe berries, with citrate comprising roughly two thirds of the total acid content. The maximum extractable catalytic activities of key enzymes in the relevant metabolic pathways were also quantified during berry development, in an attempt to elucidate possible correlations. Sucrose synthase activity was very low or undetected, while neutral invertase activity was relatively high, suggesting that sucrose synthase does not make a significant contribution to sink strength in these cultivars. Three enzymes, invertase, pyrophosphate-dependent phosphofructokinase (PFP) and fructose-1,6-bisphosphatase (FBPase), showed distinct activity patterns during development and may affect sugar and acid accumulation and sugar composition of strawberries.     

Immobilization of Invertase on Mesoporous Silicas to Obtain Hyper Active Biocatalysts

Comparative studies of invertase immobilized on two mesoporous cellular foams (MCF) and two supports of SBA-15 type with different pore sizes, using three highly recommended methods of protein bonding showed that activity of aminated MCF-based biocatalysts is significantly higher than that of SBA-15 counterparts, with the same textural properties. Specific activities of invertase covalently attached via GLA spacer was about 73–111% of a native enzyme. A unique microenvironment of functionalized cage-like pores of aminated MCFs is deemed to be responsible for this ultra high activity of invertase.     

施肥对黑土农田土壤全碳、微生物量碳及土壤酶活性的影响

通过对吉林省德惠市中层黑土农田6年定位施肥试验研究表明,施有机肥、秸秆还田 有机肥、秸秆还田 化肥和半量有机肥 化肥处理较单施化肥、未施肥和休闲裸地处理提高了土壤全碳、全氮含量,降低了土壤碳氮比。对玉米播前、抽穗期和收获期土壤微生物量碳测定表明,抽穗期土壤微生物量碳最高,处理间差异显著,且不同采样时期的土壤微生物量碳与土壤全碳含量之间呈高度正相关。对土壤酶活性研究表明,施有机肥较单施化肥、未施肥和休闲裸地处理显著地提高了土壤脲酶、转化酶和过氧化氢酶活性;单施化肥处理对土壤转化酶和过氧化氢酶具有一定的抑制作用;而有机肥、秸秆和化肥之间的配合施用处理的土壤酶活性表现不一致。相关分析显示土壤微生物量碳与土壤酶活性呈显著或极显著正相关。该研究结果表明,长期向中层黑土农田增施有机物,不仅提高了土壤全碳和土壤微生物量碳的含量,而且也提高土壤酶活性,有利于提高土壤养分转化效率,可使黑土质量向健康方向发展。     

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     

Purification and characterisation of Saccharomyces cerevisiae external invertase isoforms

Four external invertase isoforms (EINV1, EINV2, EINV3 and EINV4) from Saccharomyces cerevisiae were highly purified by isoelectric precipitation, ethanol precipitation, ion-exchange on QAE-Sephadex and gel filtration using Sephacryl S-200. Unlike previously published procedures for external invertase purification, a specially designed step elution was applied on QAE-Sephadex which enabled the separation of four isoforms. The isoforms have the same molecular mass and catalytic properties: K_m for sucrose (25.6 mM), pH optimum (3.5–5.0) and temperature optimum (60 °C), but they exhibit significant difference in pI values, thermal stability and chemical reactivity. Deglycosylation studies showed that the observed differences between isoforms arise from posttranslational modifications. Results showed that external invertase is a mixture of at least four isoforms, but in order to improve the efficiency of food industry processes, only the most stable isoform (EINV1) should be purified and utilised. Substantially different chemical reactivity of the isoforms could be used to improve the yield of covalent immobilization procedures.     

Partition coefficient prediction of Baker's yeast invertase in aqueous two phase systems using hybrid group method data handling neural network

A hybrid GMDH neural network model has been developed in order to predict the partition coefficients of invertase from Baker's yeast. ATPS experiments were carried out changing the molar average mass of PEG(1500–6000 Da), p H(4.0–7.0), percentage of PEG(10.0–20.0 w/w), percentage of MgSO_4(8.0–16.0 w/w), percentage of the cell homogenate(10.0–20.0 w/w) and the percentage of MnSO_4(0–5.0 w/w) added as cosolute. The network evaluation was carried out comparing the partition coefficients obtained from the hybrid GMDH neural network with the experimental data using different statistical metrics. The hybrid GMDH neural network model showed better fitting(AARD = 32.752%) as well as good generalization capacity of the partition coefficients of the ATPS than the original GMDH network approach and a BPANN model. Therefore hybrid GMDH neural network model appears as a powerful tool for predicting partition coefficients during downstream processing of biomolecules.     

转化酶水解蔗糖加料烘焙对白肋烟品质的影响

为提高白肋烟的处理效果,进行了转化酶水解蔗糖和柠檬酸水解蔗糖的加料烘焙试验,并进行了加料烘焙前后烟叶还原糖、总糖、烟碱、钾、氯、氨基酸、中性和碱性香味成分分析及卷烟评吸。结果显示:①与加酸转化糖里料相比,采用加酶转化糖里料对烘焙后白肋烟中的常规化学成分影响基本相同,总氨基酸含量显著降低,碱性香味成分增多;②加酶转化糖里料的白肋烟的感官质量总体上比加酸转化糖的略有改善,其中香气质和香气量有较大改善,烟气浓度和凝聚性有一定程度的改善,白肋烟香和刺激性增强,杂气、干燥感、舒适程度等指标差异不大。酶转化糖可以替代酸转化糖进行白肋烟的加料。     

Purification and characterization of soluble invertases from suspension-cultured bamboo (Bambusa edulis) cells

An alkaline invertase (IT I) and an acid invertase (IT II) were purified from the soluble fraction of suspension cultured bamboo cells. Both purified invertases were homogeneous as examined by SDS–polyacrylamide gel electrophoresis (SDS–PAGE) and were identified as β-fructofuranosidases able to attack the β-fructofuranoside from the fructose end. With respect to sucrose hydrolysis, the optimal pHs were 7.0 and 4.5 for IT I and IT II, respectively. The Km’s were 10.9 and 3.7 mM. The IT I and IT II molecular masses were 240 and 68 kDa, respectively, as estimated by gel filtration. The isoelectric points were 4.8 and 7.4. IT I was a homotetrameric enzyme activated by bovine serum albumin (BSA). IT II was a monomeric enzyme activated by BSA, concanavalin A (ConA) and urease. Both isoforms were significantly inhibited by heavy metal ions Ag⁺ (5 mM) and Hg²⁺ (1 mM), and mercaptide forming agent ρ-chloromercuribenzoic acid (PCMB; 0.5 mM).     

辐射聚合丙烯酰胺固定面包酵母

本文研究用~(60)Co辐射聚合丙烯酰胺,包埋具有转化酶活力的酵母细胞的方法。丙烯酰胺作为载体,NN‘-甲撑双丙烯酰胺作为交联剂。探讨了固定化的一些条件,如辐照剂量、聚合温度、酵母和交联剂的用量对酶活力的影响。试验发现,辐照剂量高达2×10~6rad,对酶活力没有影响。由最适条件所制得的固定化细胞既有弹性又有硬度,并且显示较高的活力回收,重复10次反应后酶活力保持不变。