PbPH5,an H^(+)P-ATPase on the tonoplast,is related to malic acid accumulation in pear fruit

Organic acid content is one of the most important factors influencing fruit flavors.The predominant organic acid in most pear cultivars is malic acid,but the mechanism controlling its accumulation remains unclear.In this study,by comparing gene expression levels and organic acid contents,we found that the expression of PbPH5,which encodes a P_(3A)-ATPase,is highly correlated with malic acid accumulation in four different pear species,with correlation coefficients of 0.932**,0.656*,0.900**,and 0.518*(*,P<0.05;**,P<0.01)for Pyrus bretschneideri Rehd.,P.communis Linn.,P.pyrifolia Nakai.,and P.ussuriensis Maxim.,respectively.Moreover,the overexpression of PbPH5 in pear significantly increased the malic acid content.In contrast,silencing PbPH5 via RNA interference significantly decreased both its transcript level and the pear fruit malic acid content.A subcellular localization analysis indicated that PbPH5 is located in the tonoplast.Additionally,a phylogenetic analysis indicated that PbPH5 is a PH5 homolog gene that is clustered with the Petunia hybrida,Malus domestica,and Citrus reticulata genes.Considered together,these findings suggest that PbPH5 is a functionally conserved gene.Furthermore,the accumulation of malic acid in pear fruit is at least partly related to changes in the PbPH5 transcription levels.

Trace determination of cobalt ion by using malic acid-malonic acid double substrate oscillating chemical system

A novel kinetic method for determination of trace amounts of cobalt ion was proposed and validated. The method is based on adding malic acid into classical Belousov-Zhabotinskii （B-Z） oscillating chemical system to form a double substrate one. The results showed that when the concentration of cobalt ion was in the range of 5.27× 10^-8 to 5.37 × 10^-12 mol L^-1, the change of the oscillating period was directly proportional to the negative logarithm of cobalt ion concentration. The sensitivity and precision of the developed method were quite satisfactory. The limit of detection was down to 5.20 × 10^-13 mol L^-1 which was a highest sensitivity found for determination of metal ions using oscillating chemical reaction so far. Some factors influencing the determination were also examined. The method has been successfully used to determine cobalt ion in vitamin B12 injection.

A convenient synthesis of methyl 2-hydroxy-4-oxo-4-(substituted phenyl) butanoates from malic acid

A synthesis of methyl 2-hydroxy-4-oxo-4-（substituted phenyl）butanoates has been accomplished using Friedel-Crafts acylation of substituted benzenes with 2-acetoxybutanoyl chloride. The key feature of this strategy is that various 4-arylbutanoates can be obtained conveniently, which are important intermediates for the synthesis of biologically active compounds including ACE inhibitors.？2009 Yong Sup Lee. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

Complexation of Molybdenum (Ⅵ) with R-and S-malic Acid, The Crystal Structure of (NH_4)_4[(MoO_2)_4O_3(R-mal)_2]·6H_2O

Ammonium tetramolybdate ( NH4 )4[(MoO2)4O3 (R-mal)2]·6H2O(mal= malic acid)and (NH4)4[(MoO2)4O3(S-mal)2)·6H2O have been prepared by the reaction of R and S-malic acid with ammonium heptamolybdate, respectively. The two complexes are enantiomeric and isomorphous. The complex of Risomer crystallizes in monoclinic space group C2 with unit cell parameters:a = 14. 591 (2 ), b = 10. 108(2), c=11. 464(2), β=121. 48(1)°, V=1441. 9(4),Z=2, Fω=1002. 13, Dc=2. 308, Dm=2. 3 g/cm3. F(000) = 988, MoKα radiation (λ=0. 71073 ), μ (MoKα) = 17. 7 cm-1. Final R=0. 027 for 1491 reflections with I>2. 5 σ(I). Analysis of molecular structure reveals that the complex anion contains a (Mo2O5)O(Mo2O5)core with the bridging oxo-group passing through two-fold axis. Each molybdenum is hexacoordinated , and the malate ligand is quadridentated to two molybdenum atoms by the deprotonated hydroxy-, α- and β-carboxylate groups. Principal dimensions are:[Mo=O(t)]av, 1. 705(6); [Mo-O(b)]av, 1. 920(6); Mo-O(hydroxy). 1. 925(6);Mo-O(α-carboxy), 2. 226 (6), and [Mo-O(β-carboxy)av, 2. 296 (5).Lattice water is distributed throughout the cell and together with various types of the anion oxygens, making H-bonding.

Preliminary Study on Lipase-catalyzed Synthesis of Polyesters Containing L-Malic Acid Units

Terpolymer of 1, 8-octanediol, adipic acid, and L-malic acid was synthesized via a lipase-catalyzed direct polycondensation. The products were characterized by GPC and 1H NMR. The results indicated that the molecular weight of the prepared polymers decreased with increasing L-malic acid content in the monomer feed ratio, and that change in the L-malic acid content from 0 to 20 mol % did not remarkably influenced on the molecular weight distribution Mw /Ma of the prepared samples. The 1H NMR spectra of the obtained copolymer samples showed that hydroxyl groups of L-malic acid did not take part in the polymerization reaction.

PREPARATION AND CELL COMPATIBILITY OF FUNCTIONALIZED BIODEGRADABLE POLY(DL-LACTIDE-co-RS-β-MALIC ACID)

In order to create a functionalized biodegradable polymer for vascular tissue engineering application,poly(DL- lactide-co-RS-β-malic acid)(PDLLMAc)was synthesized.PDLLMAc was obtained after hydrogenolysis of poly(DL- lactide-co-RS-β-benzyl malolactonate)(PDLLMA),which was from the ring-opening polymerization of DL-lactide(DLLA) and RS-β-benzyl malolactonate(MA)using stannous octoate as catalyst.The copolymers were characterized by ~1H-NMR, FTIR,GPC and DSC.The tensile strength and water uptake of the copoly...

Effect of Phenolic Compounds on the Growth and L-Malic Acid Metabolism of Oenococcus oeni

The effect of some phenolic compounds recurrent in wines on technological features of Oenococcus oeni was studied in order to individuate those strains to be utilized as starter in the deacidification of aged red wines. For this purpose, the growth and the L-malic acid metabolism of 100O. oeni strains, previously isolated from different wines, was assayed in a synthetic medium added with ethanol, malic acid and phenol carboxylic （gallic, caffeic, p-coumaric and ferulic） acids or flavonoids （catechin and quercetin） at different concentrations. Results evidenced a different sensitivity of strains to each assayed compound. All the compounds restrained or stimulated the growth of 57 and 11 strains respectively, while no effect was detected on 6 strains. The remaining 26 strains showed a different behaviour： all were restrained by ferulic acid and stimulated by gallic acid and catechin. As for caffeic acid, 17 out of 26 strains were restrained, while 9 strains were stimulated. The main result obtained in this study was the establishment of a relationship between the effect of phenolic compounds on the O. oeni growth and the behaviour of the malolactic fermentation. This study may enrich the selection criteria of strains for the deacidification of aged red wines.

Highly selective/enantioselective Pt-ReOx/C catalyst for hydrogenation of L-malic acid at mild conditions

The catalyst preparation strategy was based on a strict introduction sequence of rhenium and platinum precursors and their strong interaction with carbon support resulted in the formation of 0.5 nm Pt-Re Ox species of atomic dispersion, where platinum is metallic, while monolayer rhenium is partially oxidized（Re2＋）. The reaction kinetics was studied taking into account the process of L-malic acid association leading to the formation of inactive cyclic oligomeric species. High TOFs（ca. 50 h-1）, selectivities（ca. 99%）and stability of Pt-Re Ox/C catalysts in aqueous-phase hydrogenation of L-malic acid, which are close to those of the homogeneous pincer type complexes, were revealed at mild conditions（T = 90–130 ℃）. Taking into account that（i） hydrogenation reaction occurred 2–3 orders of magnitude faster than its racemization and（ii） association of L-malic acid dominates at low temperatures and in a concentrated solution,special reaction conditions that allow obtaining chemically and optically（ee 〉 99%） pure（S）-3-hydroxy-γ-butyrolactone and（S）-1,2,4-butanetriol were found. Basing on HAADF-STEM, EDX, XPS, and kinetic studies, the structure of active species and basic reaction pathways are proposed.

Equilibrium of Extraction of Succinic, Malic, Maleic and Fumaric Acids with Trioctylamine

Extraction equilibrium features of succinic acid, malic acid,maleic acid and fumaric acid were investi- gated systematically withtrioctylamine (TOA) in chloroform, 4-methyl-2-pentanone (MIBK) and1-octanol. Fourier transform-infrared (FTIR) spectroscopic analysisof organic samples loaded with the acid shows that amine forms 1:1complex of ion-pair association with succinic acid, malic acid andmaleic acid, and 1:1, 2:1 complex of ion-pair association withfumaric acid. It is proposed that the complex forms depend on thesecond dissociation constant of the dibasic acid, pK_a2.

Separation and Structure of Chiral S-Malic Acid Hydrate

Crystals of the chiral malic acid hydrate (C_4H_6O_5·H_2O) were unexpectedlyobtained from an ethanol/water solution containing racemic D, L-malic acid and L-asparagine. Thecrystal belongs to orthorhombic space group P2_12_12_1 with cell dimensions of a = 0.5576(1), b =0.9818(2), c = 1.1793(2) nm, V = 0.6455(2) nm^3, Z = 4, μ(Mo K_a) =0.152 mm^(-1), F(000) = 320, D_c= 1.565 g·cm^(-3), R=0.051, wR = 0.136 for 657 observed reflections [ I > 2σ(I) ]. Thesignificant difference in bond distances for carboxyl groups suggests that the crystal consist ofmalate anion and protonized crystalline water. This is a report for direct separation of racemicmixture, i.e., without the formation of a molecular complex of raceme with a chiral separatingreagent.

低分子量有机酸(苹果酸)对方解石-氟的吸附/沉淀反应影响

低分子量有机酸常对矿物的表面反应(吸附/沉淀)产生影响,从而影响矿物的溶解、矿化等过程,进而影响环境地球化学进程中元素的迁移稳定性。苹果酸作为一种广泛存在于自然界中的有机酸,是植物通过代谢过程分泌的副产品。通过批量平衡法开展了苹果酸对方解石-氟的吸附/沉淀反应的影响研究,旨在深入理解有机酸在地球化学过程中的作用。结果表明:①初始pH值为7.7条件下,对于低浓度氟(≤5 mg·L^(-1)),随着苹果酸浓度的升高,其对氟去除的抑制作用呈增强趋势;对于中、高浓度氟(25或60 mg·L^(-1)),氟去除主导机制为CaF 2沉淀反应,苹果酸的抑制作用对其影响不大,但其表面吸附反应导致pH值升高和钙浓度下降。②初始pH值为8.3条件下,对于低浓度氟(≤20 mg·L^(-1)),苹果酸对氟去除仍有抑制作用;苹果酸与Ca 2+的络合反应促使pH值和钙浓度上升;对于中浓度氟(60 mg·L^(-1)),氟去除主导机制为CaF 2沉淀反应,20 mg·L^(-1)苹果酸已对其产生抑制作用,随着苹果酸浓度继续升高,pH值先降后升,钙浓度持续上升,彰显了苹果酸络合反应的效应;对于高浓度氟(100 mg·L^(-1)),100 mg·L^(-1)苹果酸能极大抑制CaF 2沉淀反应,对应的pH值未超过无苹果酸时,对应的钙浓度仍低于其空白背景值,暗示了CaF 2沉淀反应的主导性。③初始pH值为8.7条件下,对于低浓度氟(≤5 mg·L^(-1)),苹果酸对氟去除的抑制作用有所减弱,但其络合反应产生的效应十分显著,导致pH值和钙浓度上升;对于高浓度氟(240 mg·L^(-1)),氟去除主导机制为CaF 2沉淀反应,该反应随着苹果酸浓度的升高而受到抑制,对应的钙浓度不断上升彰显苹果酸络合反应的效应,而pH值的下降也表明了CaF 2沉淀反应的主导性。本研究深入探讨了氟元素在方解石矿物界面上的吸附、迁移和转化过程,为理解氟的迁移稳定性提供了新的视角和理论基础,同时对富含有机酸环境中方解石矿物的稳定性评估具有重要意义,也为氟在其他矿物上的迁移转化研究提供借鉴。

外源有机酸对复垦土壤中重金属Cr的活化效果研究

为了探究低分子有机酸(柠檬酸和苹果酸)对Cr污染土壤中Cr形态分布的影响,以淮南市潘一矿煤矸石山周边的复垦土壤为研究对象,通过模拟不同浓度Cr污染土壤,分析柠檬酸(CA)和苹果酸(MA)对土壤中Cr赋存形态的影响机制。结果表明:(1)随着外源Cr浓度的增加,土壤中Cr的活性有效提高。(2)柠檬酸和苹果酸的加入均可增加土壤中可交换态Cr的占比,即Cr的有效性得到提高,且随着柠檬酸和苹果酸浓度的升高,对土壤中Cr活化效果持续上升,柠檬酸的活化效果优于苹果酸。(3)随着老化时间的延长,有机酸对Cr的活化效果会减弱,弱结合态Cr将逐渐向强结合态Cr转化。因此,土壤重金属Cr污染修复中,可以考虑选用活化效果较好的柠檬酸作为修复辅助剂,来提升土壤Cr修复效率。研究结果可为矿区复垦土壤重金属Cr污染修复提供指导。

一株产L-苹果酸黑曲霉菌株的诱变筛选及发酵培养基优化

为筛选一株产L-苹果酸能力强的黑曲霉菌株,通过紫外诱变与高浓度放线菌酮迭代诱变的方法,将野生型菌株诱变为一株产L-苹果酸能力强的黑曲霉菌株,并对其种子或发酵培养基成分进行优化。结果表明,诱变所得一株产L-苹果酸能力强的黑曲霉CGMCC NO.40550菌株,其摇瓶发酵时种子培养基最适葡萄糖浓度为60 g/L、最适氮源为(NH_4)_2SO_4 4.95 g/L,黑曲霉菌球形成最优转速220 r/min;发酵培养基最适葡萄糖和最适(NH_4)_2SO_4浓度分别为180 g/L和4.95 g/L,Cu SO_4·5H_2O为其生产苹果酸最佳微量元素,最适添加量为0.065 g/L。通过单因素实验的优化,优化后的培养基更有利于L-苹果酸的合成,发酵96 h时L-苹果酸产酸量达到18.15 g/L,显著提高了245%,L-苹果酸占总酸的百分比达到71.69%。本研究为L-苹果酸的工业化生产奠定基础。

苹果酸大米淀粉酯的制备及其对面条品质特性的改善

为探究不同取代度(Degree of Substitution,DS)对苹果酸大米淀粉酯理化特性及对面条品质的影响,制备不同取代度(DS=0~0.603)苹果酸大米淀粉酯,考察了苹果酸大米淀粉酯溶解度、溶胀度、糊化特性和流变特性,以及对面条蒸煮特性、质构特性和感官特性的改善作用。结果表明,苹果酸淀粉酯的红外光谱分析显示在光谱波数约1720 cm-1处出现一个新的吸收峰。淀粉发生酯化反应后溶解度显著升高(P<0.05),但溶胀度有所下降。糊化和流变实验结果表明,低酯化淀粉(DS=0.141~0.442)的加入显著降低(P<0.05)了淀粉的峰值黏度,降低了苹果酸淀粉酯的储能模量G′和损耗模量G″,高酯化淀粉(DS=0.603)的添加不利于改善苹果酸淀粉酯的糊化和流变特性。利用相关性分析对苹果酸淀粉酯部分理化指标(溶解度、溶胀度和糊化特性)和面条的品质(蒸煮特性、质构特性和感官特性)进行了综合分析。苹果酸大米淀粉酯可以显著提升面条的整体品质,在本实验中,取代度为0.442时,小麦粉制备的面条品质较佳。

大豆铝激活苹果酸转运家族基因GmALMT8的鉴定与功能验证

鲜食风味是影响菜用大豆食味品质的关键因素,其形成与有机酸有着密切的关联,研究有机酸合成机制对于菜用大豆的品质改良具有重要的实际意义。本研究利用大豆毛状根系统,探究与苹果酸含量显著相关的候选基因GmALMT8、GmIF7GT5和GmAP在调控苹果酸含量方面的功能,结果表明:在GmALMT8-OE毛状根中,GmALMT8基因表达量与苹果酸的含量均显著高于空载对照毛状根,而GmIF7GT5和GmAP毛状根中苹果酸的含量无显著变化。鉴于已经报道的ALMT家族基因的苹果酸转运功能,推测大豆中GmALMT8基因可能具有相似的功能,在调控苹果酸含量方面发挥重要作用。为验证GmALMT8-OE毛状根中苹果酸含量的变化是否由GmALMT8基因表达的改变所引起,本研究采用蘸花法在拟南芥中过表达GmALMT8基因。与阳性毛状根中苹果酸测定的结果相类似,过表达GmALMT8显著提高了T2代转基因拟南芥株系种子中的苹果酸含量,进一步证明GmALMT8的稳定表达能够提高苹果酸含量,明确GmALMT8基因在大豆中具有调控苹果酸含量的生物学功能,丰富了大豆有机酸的理论研究,对菜用大豆优质育种具有参考价值。

日粮添加生物法苹果酸在中试条件下对肉鸡生产的影响

本试验旨在评估生物法苹果酸产品在肉鸡大群试验中的促生长和肉质改善效果,为生物法苹果酸在肉鸡饲粮中的推广应用提供数据支撑。选用1000只1日龄的爱拔益加肉仔鸡公雏,随机分为2个处理,每个处理10个重复,每个重复50只鸡。对照组饲喂玉米-豆粕型基础日粮,处理组在基础日粮中添加0.8%生物法苹果酸。试验持续42 d。结果表明:日粮添加0.8%生物法苹果酸对肉鸡采食量、死淘率无显著影响(P> 0.05),但能通过增加试验后期(22~42 d)平均日增重(7.53%)(P=0.001)和降低料重比(4.47%)(P=0.002)有效增加肉鸡42 d出栏重(4.93%)(P <0.001);在屠宰性能和肉品质方面,日粮添加苹果酸对全净膛率、胸肌率、腿肌率、腹脂率、胸肌肉色和pH均无显著影响(P> 0.05),但能显著降低胸肌肉的滴水损失(11.31%)(P <0.001)。日粮添加0.8%苹果酸可以在中试条件下有效提高肉鸡的生长性能,增强鸡肉保水性,改善肉品质。因此,本研究推荐生物法苹果酸作为饲料添加剂,在集约化养殖的背景下应用于肉鸡饲粮。

重组大肠杆菌全细胞转化苹果酸合成丙酮酸

拟建立一条以苹果酸为原料,通过大肠杆菌全细胞转化获得丙酮酸的合成途径。以大肠杆菌BL21(DE3)为宿主,在pET28a上共表达内源的苹果酸酶(ME)和来自博伊丁假丝酵母(Candida boidinii)的醛糖还原酶(AR),并对细胞浓度、底物浓度、温度和pH值等催化关键因素进行研究。共表达后大肠杆菌苹果酸酶和博伊丁假丝酵母醛糖还原酶酶活分别为(2.8±0.21),(3.1±0.34)U/mL。适宜的催化条件为:细胞浓度OD600nm值为30,底物苹果酸质量浓度为30 g/L,木糖和苹果酸物质的量比为1.0,温度为40℃,反应体系pH值为7.8,丙酮酸产量最高可达23.16 g/L。本研究为生物法合成丙酮酸提供了一种新的方法。

产聚苹果酸出芽短梗霉的筛选、鉴定及发酵培养基优化

从自然土壤样本中筛选得到一株产聚苹果酸(PMLA)菌株,该菌株经形态学观察和分子进化分析鉴定为出芽短梗霉(Aureobasidium pullulans)。筛选菌株同时具有聚苹果酸、普鲁兰多糖和liamocin胞外聚合物的合成能力。通过单因素和响应面试验对该菌种产PMLA的发酵培养基组分进行优化,获得较优培养基组分为葡萄糖130 g/L、硝酸钠5 g/L、玉米浆1.22 g/L、氯化钾0.5 g/L、磷酸二氢钾0.1 g/L、七水硫酸镁0.2 g/L、碳酸钙30 g/L,采用该培养基获得聚苹果酸的最大产量为25.82 g/L,比优化前提高了70.72%。

葡萄籽原花青素和苹果酸对面包抗老化的影响及机制分析

面包在贮藏过程中因老化会出现硬度变大、水分流失、粗糙掉屑等问题,该研究以葡萄籽原花青素(grape seed proanthocyanidins,GSP)和苹果酸(malic acid,MA)为主要原料,研究二者协同对面包抗老化的影响。采用快速黏度仪(rapid visco analyzer,RVA)测定了面粉的糊化特性,对贮藏期间面包水分含量、硬度、水分分布的变化进行了测定,利用RVA、差示扫描量热仪(differential scanning calorimeter,DSC)和X射线衍射仪(X-ray diffractometer,XRD)对贮藏期间淀粉的回生进行了研究。结果表明,GSP和MA降低了贮藏期间面包水分含量的下降速率,在0.3%(质量分数,下同)GSP中复配0.1%MA、0.3%MA和0.5%MA的面包硬度相对于0.3%GSP面包分别降低了9.34%、10.49%、8.04%。面包贮藏5 d的DSC和XRD数据表明GSP和MA的添加显著降低了淀粉的回生焓值和结晶度,在0.3%GSP中复配0.1%MA、0.3%MA、0.5%MA制作的面包其结晶度相较于0.3%GSP分别下降了33.54%、36.08%和34.18%。综上所述,GSP和MA对延缓面包的老化起到协同作用。

野酿2号毛葡萄果实发育进程中苹果酸代谢规律及相关基因表达分析

【目的】分析野酿2号毛葡萄果实发育进程中苹果酸代谢规律及相关基因表达情况,为提高野酿2号毛葡萄的酿造品质及优质葡萄酒生产提供参考依据。【方法】以5年生野酿2号毛葡萄为试验材料,于盛花后5~17周采集不同生长发育阶段果实,测定果实单粒重、pH、可溶性固形物和可滴定酸含量,采用高效液相色谱法测定果实中苹果酸、柠檬酸和酒石酸含量;使用酶标仪测定苹果酸代谢关键酶[苹果酸脱氢酶(NAD-MDH)、NADP-苹果酸酶(NADP-ME)和磷酸烯醇式丙酮酸羧化酶(PEPC)]活性;采用实时荧光定量PCR检测苹果酸代谢相关基因PEPC、MDH和ME的相对表达量,并分析3个基因与苹果酸含量的相关性。【结果】野酿2号毛葡萄果实单粒重在转色期前增长较快,转色期的果实pH、可溶性固形物、苹果酸含量分别是盛花后5周的1.03、1.36和2.38倍,盛花后11周可滴定酸含量达峰值,为35.68 mg/g。盛花后5~17周,野酿2号毛葡萄果实中酒石酸和柠檬酸含量变化不明显,而苹果酸含量变化幅度较大。盛花后8周,NAD-MDH和NADP-ME活性最高,分别为736.06和453.50 nmol/(g·min),幼果期PEPC活性较高。转色期果实中MDH基因的相对表达量是盛花后5周的1.62倍,而PEPC和ME基因的相对表达量均低于盛花后5周。相关分析结果表明,苹果酸含量与MDH基因呈极显著正相关(P<0.01),与PEPC基因呈显著正相关(P<0.05,下同),与ME基因呈显著负相关。【结论】野酿2号毛葡萄苹果酸含量变化是影响总酸变化趋势的主要因素,转色期是苹果酸含量由积累到降低的转折期,苹果酸含量受相关基因MDH、PEPC和ME表达调控。