Effect of intrinsic and extrinsic factors on the interaction of plant pectin methylesterase and its proteinaceous inhibitor from kiwi fruit

A proteinaceous pectin methylesterase inhibitor (PMEI) was isolated from kiwi fruit (Actinidia chinensiscv. Hayward) and purified by affinity chromatography on a cyanogen bromide (CNBr) Sepharose 4B-orange PME column. The optimal pH of banana PME activity was 7.0, whereas that for carrot and strawberry PME activity was 9.0. The optimal pH for the binding between kiwi fruit PMEI and these PMEs was 7.0. The kiwi fruit PMEI has a different affinity for PME depending on the plant source. The inhibition kinetics of kiwi fruit PMEI to banana and strawberry PME followed a noncompetitive type, whereas that to carrot PME followed a competitive type. The kiwi fruit PMEI was mixed with banana, carrot, and strawberry PME to obtain PMEI-PME complexes, which were then subjected to thermal (40-80 degrees C, atmospheric pressure) or high-pressure (10 degrees C, 100-600 MPa) treatment. Experimental data showed that the PMEI-PME complexes were easily dissociated by both thermal and high-pressure treatments.     

猕猴桃汁维生素C降解动力学的研究

以猕猴桃原汁为试材,采用控制温度、调节pH值以促使维生素C氧化降解的方法,探讨猕猴桃汁中维生素C的降解动力学,为控制猕猴桃汁加工中维生素C损失提供理论依据。结果表明：猕猴桃汁中的维生素C降解十分复杂,存在多种降解途径,但主要是有氧降解;其还原型维生素C不经氧化型维生素C生成其他产物的不可逆途径可能存在,但不占主导地位。猕猴桃汁中维生素C无氧降解速度比有氧降解速度慢,温度升高,维生素C有氧降解速度加快,无氧降解速度变化不明显。     

Aromatic characterization of pot distilled kiwi spirits

This study contributes fundamental knowledge that will help to develop a distillate of kiwi wine, made from kiwis of the Hayward variety grown in the southwest of Galicia (Spain). Two yeast strains, L1 (Saccharomyces cerevisiae ALB-6 from the EVEGA yeast collection) and L2 (S. cerevisiae Uvaferm BDX from Lallemand) were assessed to obtain a highly aromatic distillate. The kiwi spirits obtained were compared with other fruit spirits, in terms of higher alcohols, minor alcohols, monoterpenols, and other minor compounds, which are relevant in determining the quality and taste of the kiwi spirits. It was found that the kiwi juice fermented with yeast L1 produced a more aromatic distillate. In addition, kiwi distillates produced with both yeasts had the same ratio of trans-3-hexen-1-ol and cis-3-hexen-1-ol, which is lower than that found in other fruit distillates.     

Aroma active components in aqueous kiwi fruit essence and kiwi fruit puree by GC-MS and multidimensional GC/GC-O

Gas chromatography-mass spectrometry (GC-MS) and multidimensional gas chromatography olfactometry (GC/GC-O) were utilized to study the aroma profile and the aroma active components of commercial kiwi essence and the initial fresh fruit puree. Totals of 29 and 33 components were identified and quantified in the essence and the puree, respectively. Ten components were quantified for the first time as constituents of the kiwi fruit including 3-penten-2-ol, 3-hydroxy-2-butanone, 3-methyl-2-butenal, 2-hexanol, nonanal, 3-methyl-1-butanol, 2-methyl-1-butanol, 3-methyl-2-butanone, 3-methyl 3-buten-2-one, and octane. Analysis of these samples by multidimensional gas chromatography-olfactometry (GC-O) allowed for the identification of >80% of the aroma active components present at level traces in this fruit. A total of 35 components appear to contribute to the aroma of kiwi fresh puree and its aqueous essence. Components described for the first time as constituents of the aroma profile in this fruit were 2-ethylfuran, 3-methyl-1-butanol, 2-cyclohexen-1-one, (E,E)-2,6-nonadienal, diethyl succinate, and hexyl hexanoate.     

Kiwellin, a modular protein from green and gold kiwi fruits: evidence of in vivo and in vitro processing and IgE binding

Kiwellin, an allergenic protein formerly isolated from green kiwi fruit, has been identified as the most abundant component of the gold kiwi species. A protein named KiTH, showing a 20 kDa band on reducing SDS-PAGE and 100% identity with the C-terminal region of kiwellin, has been identified in the extract of the ripe green species. In vitro treatment of purified kiwellin with the protease actinidin from green kiwi fruit originated KiTH and kissper, a recently described pore-forming peptide. Primary structure analysis and experimental evidence suggest that kiwellin is a modular protein with two domains. It may undergo in vivo proteolytic processing by actinidin, thus producing KiTH and kissper. When probed with sera recognizing kiwellin from green kiwi fruit, KiTH showed IgE binding, with reactivity levels sometimes different from those of kiwellin. The IgE-binding capacity of kiwellin from gold kiwi fruit appears to be similar to that of the green species.     

微波提取法对樱桃、猕猴桃和枸杞多糖特性的影响

微波提取是近年来发展迅速的天然产物提取技术之一。为考察微波提取对多糖得率、结构及生物学功能的影响,论文采用微波功率1 000 W和提取温度140℃,以不同提取时间5,10 和20 min分别对樱桃、猕猴桃和枸杞中多糖进行提取。结果表明,微波提取多糖的得率、相对分子质量和抗氧化性低于传统提取方法,随着微波提取时间延长,多糖相对分子质量逐步降低,但多糖得率与抗氧化性无显著变化;微波提取得到的多糖在单糖组成上与传统提取方法有一定的差异。该研究结果对应用微波提取水果多糖条件的选择及多糖品质控制有一定的参考价值。     

Application of wood ash compared with fertigation for improving the nutritional status and fruit production of kiwi vines

Application of wood ash can potentially improve the fertility of acid soils and the nutritional status of crops. However, there is limited information about the effectiveness of this practice with fruit trees. The application of wood ash as a fertilizer in a kiwifruit plantation—both with and without fertigation/irrigation—was compared with that of a conventional fertigation program in a completely randomized field experiment on an acid soil in northwest (NW) Spain. The effects on plant nutritional status and on fruit yield, as well as environmental effects, were evaluated over a period of 2 y. The application of wood ash led to modest increases in soil pH and extractable nutrients (phosphorus, P; calcium, Ca; magnesium, Mg; potassium, K; boron, B). However, no consistent effects in foliar nutrient concentrations were found. Ash application led to an increase of up to 45% in the number of fruits produced, which was mainly attributed to the inputs of Ca and Mg. Although moderate increases in soil available manganese (Mn) and nickel (Ni) after ash application were recorded, there were no changes in heavy‐metal concentrations in leaves or fruits. From the results of the study it can be concluded that wood ash can be used to improve the growth conditions of kiwi vines on acidic soils. Wood ash should be applied at rates adapted to the liming needs of the soil, while also taking into account the chemical composition of the ash.     

Characterization of the bound volatile extract from baby kiwi (Actinidia arguta)

The glycosidically bound volatile fraction of baby kiwi ( Actinidia arguta ) was studied. Glycosidic precursors were isolated from juice by adsorption onto an Amberlite XAD-2 column. After enzymatic hydrolysis with Rapidase AR2000, the released aglycones were analyzed by GC-MS. Alcohols, terpenoids, and benzenoids were the most abundant compound classes. Aromatic compounds and norisoprenoids showed the highest concentrations. Major compounds were 2,5-dimethyl-4-hydroxy-3(2H)-furanone (Furaneol), benzyl alcohol, 3-hydroxy-β-damascone, hexanal, and (Z)-3-hexen-1-ol. Precursors of aroma compounds including benzoic acid, cinnamic acid, and coniferyl alcohol were also found. Eugenol, raspberry ketone, and 4-vinylguaiacol were identified for the first time in the fruit of an Actinidia species. The high concentration of 2,5-dimethyl-4-hydroxy-3(2H)-furanone in bound form (95.36 μg/kg) is particularly interesting and justifies further investigation.     

Antimutagenic effect of fruit and vegetable ethanolic extracts against N-nitrosamines evaluated by the Ames test.

The inhibitory effect of nine fruit and vegetable ethanolic extracts against the mutagenicity of N-nitrosodimethylamine (NDMA), N-nitrosopyrrolidine (NPYR), N-nitrosodibutylamine (NDBA), and N-nitrosopiperidine (NPIP) was evaluated by means of the Ames test. Licorice ethanolic extract was the only one that showed an inhibitory effect (ranging from moderate to strong) against mutagenicity of all N-nitrosamines tested. This ethanolic extract showed the greatest inhibition effect against NPIP (72%), NDMA (45%), and NPYR (39%). The greatest inhibition effect (51%) of the mutagenicity of NDBA was shown by kiwi ethanolic extract. Vegetable and fruit ethanolic extracts that exhibited an antimutagenic effect (at the range 50-2000 microg/plate), in decreasing order, against NDMA and NPYR were as follows: licorice > kiwi > carrot and licorice > broccoli > pineapple > kiwi, respectively. Decreasing orders against NDBA and NPIP were, respectively, kiwi > onion > licorice = garlic > green pepper > carrot and licorice > garlic > pineapple > carrot.     

Isolation, characterization, and pectin-modifying properties of a thermally tolerant pectin methylesterase from Citrus sinensis var. Valencia

The thermally tolerant pectin methylesterase (TT-PME) was isolated as a monocomponent enzyme from sweet orange fruit (Citrus sinensis var. Valencia). It was also isolated from flower and vegetative tissue. The apparent molecular weight of fruit TT-PME was 40800 by SDS-PAGE and the isoelectric point estimated as pI 9.31 by IEF-PAGE. MALDI-TOF MS identified no tryptic-peptide ions from TT-PME characteristic of previously described citrus PMEs. TT-PME did not absolutely require supplemented salt for activity, but salt activation and pH-dependent activity patterns were intermediate to those of thermolabile PMEs. Treatment of non-calcium-sensitive pectin with TT-PME (reducing the degree of methylesterification by 6%) increased the calcium-sensitive pectin ratio from 0.01 to 0.90, indicating a blockwise mode of action. TT-PME produced a significantly lower end-point degree of methylesterification at pH 7.5 than at pH 4.5. Extensive de-esterification with TT-PME did not reduce the pectin molecular weight or z-average radius of gyration, as determined by HPSEC.     

Call count responses of North Island brown kiwi to different levels of predator control in Northland, New Zealand

North Island brown kiwi have been monitored by a standard call count index at 23 stations in Northland, New Zealand annually since 1995. An attempt is made here to determine whether there are any trends in call counts that relate to predator control and advocacy in and around each listening station. Both predator and advocacy effort were rated on a 0-3 scale at each of 23 listening stations, and kiwi calling rates analysed on a generalised linear model. An analysis of variance (ANOVA) indicated a positive response of call counts to predator control. There was a weaker relationship with advocacy, but because predator control and advocacy are correlated, it was not possible to separate their effects. Ground moisture levels had a significant positive effect on calls, while wind and rain had negative effects. Overall, the analysis suggested that this kind of management is benefitting kiwi in Northland.     

Critical values of alternative organic amendments on kiwi seedling growth

The present research was conducted to explore the impact of alternative organic amendments on physicochemical properties of soil and their critical value for kiwi seedlings growth. The experiment was laid out in completely randomized design with 3 replications and 20 seedlings per replication. The highest pH (8.0) was recorded in T3 (compost + silt) and the highest electrical conductivity (EC) of 5.91 dS/m, organic matter (24.11%), organic carbon (13.99%), organic nitrogen (1.51%) were recorded in T8 (compost + peat moss (PM) + leaf manure) and the highest soil porosity (SP) (45.95%) and lowest bulk density (0.67) were recorded in T4 (PM + garden soil (GS)). EC higher than 1.99 and SP (45.95%) were found critical and adversely affected seedling survival. The highest plant height (103 and 93 cm), number of leaves (23 and 20), number of first-order lateral roots (NFLRM) (9 and 8.5), whole seedling fresh mass (75 and 72 g), fresh root mass (FRM) (35 and 35 g), shoot fresh mass (46 and 40 g), root collar diameter (RCD) (13 and 12 mm), and shoot diameter (10 and 9 mm) were recorded in T5 (compost + GS) and T7 (compost + PM + GS). Strong and positive correlations were observed among NFLRP, RFM, and RCD (R² = 0.89 and 0.96) and all the above-ground seedling traits (R² = 0.86–0.98). It is concluded that compost is a good soil amendment for producing good quality graftable kiwi seedlings and evaluation of seedling root system architecture could be used to predict the potential and success of seedlings in the field.     

Beta-glucosidase from the grape native yeast Debaryomyces vanrijiae: purification,characterization, and its effect on monoterpene content of a Muscat grape juice

Six hundred ten yeast colonies isolated from various vineyards in Chile were screened for the presence of a beta-glucosidase activity as well as the resistance to glucose and ethanol inhibition. Among them, Debaryomyces vanrijiae was found to produce high levels of an extracelular beta-glucosidase which was tolerant to glucose (K(i) = 439 mM) and ethanol inhibitions. The enzyme (designated DV-BG) was purified to apparent homogeneity, respectively, by gel filtration, ion-exchange, and chromatofocusing techniques. Its molecular weight was 100 000, and its pI 3.0, optimum pH, and temperature activities were 5.0 and 40 degrees C, respectively, and had a V(max) of 47.6 micromol min(-)(1) mg(-)(1) and a K(m) of 1.07 mM. The enzyme was active against different beta-d-glucosides including glucosidic flavor precursors. The disaccharidic flavor precursors were not substrates for the enzyme. When added to a Muscat grape juice, the concentration of several monoterpenes increased as the consequence of its hydrolytic activity.     

基于改进YOLOv5s的自然环境下猕猴桃花朵检测方法

为实现对猕猴桃花朵的快速准确检测,该研究提出了一种基于改进YOLOv5s的猕猴桃花朵检测模型YOLOv5s＿S＿N＿CB＿CA,并通过对比试验进行了精度验证。在YOLOv5s基础上引入C3HB模块和交叉注意力(criss-cross atte ntion,CCA)模块增强特征提取能力,结合样本切分和加入负样本处理方法进一步提升模型精度。改进模型的检测精确率为85.21%,召回率为90%,模型大小为14.6 MB,交并比(intersection over union,IoU)为0.5下的均值平均精度(mAP_0.5)为92.45%,比仅进行样本缩放处理的原始YOLOv5s提高了31.91个百分点,检测速度为35.47帧/s,比原始YOLOv5s提高了34.15%。使用改进模型对自然环境下不同天气、晴天不同时段光照强度下的猕猴桃花朵进行检测,结果表明模型检测晴天、阴天下猕猴桃花朵的mAP_0.5分别为91.96%、91.15%,比原始YOLOv5s分别高出2.55、2.25个百分点;检测晴天9:00-11:00、15:00-17:00光强下猕猴桃花...     

β-葡聚糖酶的热稳定性改造及酶学性质分析

为了获得热稳定性有所提高的β-葡聚糖酶，本研究利用定向进化技术对β-葡聚糖酶基因进行改造，并对野生酶和突变酶的酶学性质进行研究。结果获得了两株热稳定性有所提高的突变体，分别命名为EGs1和EGs2；酶学性质分析结果显示：与野生酶相比，EGs1和EGs2突变酶的Tm值分别提高了3℃和5℃，达到了65.5℃和67.5℃；突变酶的最适反应温度也提高了5℃，达到了60℃；两个突变酶对地衣多糖的水解能力分别被提高28％和降低21.6％；对地衣多糖的亲和力未见改变；野生酶和EGs1突变酶的最适pH值为6.5，而EGs2突变酶的最适pH值为7.0；野生型和突变型-葡聚糖酶都有较宽的pH稳定范围，在pH 6.0-8.5的范围内放置48小时，仍保持80％以上的酶活力；遗传稳定性检验结果表明突变株的遗传稳定性良好。这一结果说明了定向进化在改善酶性质方面是比较有效的策略。     

基于Adaboost算法的田间猕猴桃识别方法

实现猕猴桃自动化采摘的关键是自然环境下果实的准确识别。为提高田间猕猴桃果实的识别效果,基于Adaboost算法,利用RGB、HSI、La*b*3个颜色空间中的1个或多个通道构建6个不同的弱分类器,用采集的猕猴桃果实和背景共300个样本点进行训练生成1个强分类器。然后选择655个测试样本点进行验证,强分类器分类精度为94.20%,高于任意弱分类器。对80幅图像中215个猕猴桃进行试验,结果表明：Adaboost算法可有效抑制天空、地表等复杂背景的影响,适合于自然场景下的猕猴桃图像识别,识别率高达96.7%。该技术大大提高了猕猴桃采摘机器人的作业效率。     

氯吡脲对贵长猕猴桃果实氨基酸和香气成分的影响

为探讨未浸果和浸果处理(氯吡脲10 mg·L^-1)对贵长猕猴桃果实产量品质、氨基酸和香气成分的影响,本试验测定了采收期果实产量参数和可食期营养品质,并采用高效液相色谱法(HPLC)和固相微萃取-色谱-质谱联用法(SPME-GC-MS)分别分析了可食期果实氨基酸和香气成分。结果表明,10 mg·L^-1氯吡脲浸果处理可有效促进贵长猕猴桃果实产量形成及干物质和可溶性固形物含量增加,显著降低果实果形指数和硬度以及维生素C、可滴定酸和可溶性蛋白含量(P<0.05),影响果实的外观和耐贮性。此外,10 mg·L^-1氯吡脲浸果极显著增加果实脯氨酸和甜味氨基酸含量(P<0.01),而果实总氨基酸、鲜味氨基酸、苦味氨基酸和芳香族氨基酸含量显著降低(P<0.05)。 贵长猕猴桃果实香气成分主要为酯类、醇类、醛类、烯烃和烷烃,共70种成分。其中,未浸果果实香气成分主要为丁酸乙酯、丁酸甲酯、苯甲酸甲酯和苯甲酸乙酯等,浸果果实主要为丁酸乙酯、苯甲酸甲酯、丁酸甲酯和(E)-2-己烯醛等。10 mg·L^-1氯吡脲浸果增加了醛类、醇类、烯烃和烷烃在果实香气成分中的占比,提高了香气的多样性。本研究为贵长猕猴桃的深加工提供了理论依据。     

Separation and characterization of a salt-dependent pectin methylesterase from Citrus sinensis var. Valencia fruit tissue

A pectin methylesterase (PME) from sweet orange fruit rag tissue, which does not destabilize citrus juice cloud, has been characterized. It is a salt-dependent PME (type II) and exhibits optimal activity between 0.1 and 0.2 M NaCl at pH 7.5. The pH optimum shifted to a more alkaline range as the salt molarity decreased (pH 8.5-9.5 at 50 mM NaCl). It has an apparent molecular mass of 32.4 kDa as determined by gel filtration chromatography, an apparent molecular mass of 33.5 kDa as determined by denaturing electrophoresis, and a pI of 10.1 and exhibits a single activity band after isoelectric focusing (IEF). It has a K(m) of 0.0487 mg/mL and a V(max) of 4.2378 nkat/mg of protein on 59% DE citrus pectin. Deblocking the N-terminus revealed a partial peptide composed of SVTPNV. De-esterification of non-calcium-sensitive pectin by 6.5% increased the calcium-sensitive pectin ratio (CSPR) from 0.045 +/- 0.011 to 0.829 +/- 0.033 but had little, if any, effect on pectin molecular weight. These properties indicate this enzyme will be useful for studying the PME mode of action as it relates to juice cloud destabilization.     

Activity, cloning, and expression of an isoamylase-type starch-debranching enzyme from banana fruit

Unripe bananas have a high content of starch (almost 20%) that is metabolized during fruit ripening with a concomitant synthesis of soluble sugars. Since starch granules are composed of amylose and amylopectin, several enzymes have to be involved in its mobilization during banana ripening, with a necessary participation of one starch-debranching enzyme (DBE) to hydrolyze the alpha-1,6-branches of amylopectin. Banana DBE seems to be an isoamylase-type enzyme, as indicated by substrate specificity and the cloning of a 1575 bp cDNA, similar to the isoamylase sequences from potato, Arabdopsis, and maize. The assays for DBE indicated only minor changes in activity during ripening, and the results of the northern and western blots with antiserum against the recombinant banana isoamylase were in agreement with the steady-state level of activity, since no significant changes in gene expression were observed. The high activity on beta-limit dextrin and the similarity to the potato isoform 3 suggest that during banana ripening the hydrolysis of alpha-1,6-linkage of amylopectin results from the activity of a pre-existing isoamylase-type debranching enzyme in coordination with other amylolitic enzymes. To the best of our knowledge, this is the first evaluation of activity and expression of a DBE from a fruit.     

Assessment of the volatile composition of juices of apricot, peach, and pear according to two pectolytic treatments

The behavior of volatile compounds according to two enzymatic treatments applied during the manufacture of fruit juice is described. More than 80 compounds were detected of a wide range of chemical families (alcohols, aldehydes, ketones, terpenoids, esters, norisoprenoids, ...). Theaspirane and alpha-isophoron were tentatively identified for the first time in apricot and peach fruits. The enzymes used, for extraction or clarification of fruit juices, modified the polysaccharides separated by molecular weight and the content of soluble polysaccharides. This could indicate differences in the fruit juice matrix, which could be related to observed changes in the volatile profile. In apricot, the enzymes enhanced the juice in terpenes and norisoprenoids as varietal compounds. In peach and pear, the enzymes used did not favor the amount of lactones and decadienoate esters, the character impact compounds, respectively.