Enzymatic synthesis of structured phenolic lipids by incorporation of selected phenolic acids into triolein

The synthesis of structured phenolic lipids by lipase-catalyzed transesterification of selected phenolic acids, including p-hydroxyphenyl acetic, p-coumaric, sinapic, ferulic and 3,4-dihydroxybenzoic acids, with triolein was investigated. The highest enzymatic activity (248?nmol esterified phenolic acid/g solid enzyme/min) and bioconversion (62%) was obtained for the transesterification of p-hydroxyphenyl acetic acid with triolein. In addition, the transesterification of p-coumaric with triolein resulted in a higher enzymatic activity (87?nmol esterified phenolic acid/g solid enzyme/min) and bioconversion (46%) than those obtained for the transesterfication of ferulic and sinapic acids. The results also showed that using p-hydroxyphenyl acetic, p-coumaric and ferulic acids as substrate, the maximum bioconversion of phenolic monoacylglycerols was close to that of phenolic diacylglycerols. Although p-coumaric acid had very low radical scavenging activity (2%) compared to that of ferulic acid (62%), the p-coumaroylated lipids demonstrated a higher scavenging potency (16%) than that of the feruloylated one (10%).     

Enzymatic synthesis of hydrophilic and hydrophobic derivatives of natural phenolic acids in organic media

The enzymatic esterification of natural phenolic antioxidants such as cinnamic acid and benzoic acid derivatives, with aliphatic alcohols, monosaccharides as well as alkylglucosides, using various lipases and esterases in non-aqueous media, was investigated. Reaction rate and esterification yield seems to be linked to the structural characteristics of the substrates (aromatic acids and alcohols or sugars) used.     

Lipase-catalyzed synthesis of structured phenolic lipids in solvent-free system using flaxseed oil and selected phenolic acids as substrates

Structured phenolic lipids (PLs) were obtained by lipase-catalyzed transesterification of flaxseed oil, in a solvent-free system (SFS), with selected phenolic acids, including hydroxylated and/or methoxylated derivatives of cinnamic, phenyl acetic and benzoic acids. A bioconversion yield of 65% was obtained for the transesterification of flaxseed oil with 3,4-dihydroxyphenyl acetic acid (DHPA). However, the effect of the chemical structure of phenolic acids on the transesterification of flaxseed oil in SFS was of less magnitude as compared to that in organic solvent system (OSS). Using DHPA, the APCI-MS analysis confirmed the synthesis of monolinolenyl, dilinolenyl, linoleyl linolenyl and oleyl linolenyl dihydroxyphenyl acetates as phenolic lipids. A significant increase in the enzymatic activity from 200 to 270 nmol of PLs/g solid enzyme/min was obtained upon the addition of the non-ionic surfactant Span 65. However, upon the addition of the anionic surfactant, sodium bis-2-ethylhexyl sulfosuccinate (AOT), and the cationic one, hexadecyltrimethylammonium bromide (CTAB), the enzymatic activity was decreased slightly from 200 to 192 and 190 nmol of PLs/g solid enzyme/min, respectively. The results also showed that the increase in DHPA concentration from 20 to 60 mM resulted in a significant increase in the volumetric productivity (P(V)) from 1.61 to 4.74 mg PLs per mL reaction mixture per day.     

Enzymatic synthesis of aldonic acids

Several aldonic acids (D-mannonic, D-galactonic, D-xylonic, 2-deoxy-D-arabinohexonic (2-deoxy-D-gluconic)) were prepared on a scale of several grams by a simple oxidation catalyzed by glucose oxidase in pure water.     

Quenching of fluorescence of phenolic compounds and modified humic acids by cadmium ions

The interaction of a number of phenolic compounds, being ‘model fragments’ of humic acids, with cadmium ions was investigated. The fluorescence quenching method was used to determine the complexation constants of these compounds with cadmium ions. It was established that bonding of phenolic compounds by cadmium ions at рН 7 is weak and reaches a maximum value of 15% for interaction with resorcinol. It was demonstrated that modification of humic acids by the mechanoactivation method increases by three times bonding of cadmium ions, which is caused by strengthening the acid properties of carboxyl and hydroxyl groups at the aromatic ring. Copyright © 2016 John Wiley & Sons, Ltd.     

酚酸类物质的化感作用研究进展

酚酸类物质是普遍存在于高等植物组织并与植物生长密切相关的次级代谢产物。几十年来,人们对酚酸类化合物的认识逐渐加深,但关于其在生物学、生态学以及农业上的作用机制仍不是很清楚。因此,进一步了解这些生物分子将有助于生态系统的维持与保护。重点介绍了酚酸类物质的来源及化感作用,微生物对酚酸类物质的降解机理,代谢途径及相应分子水平的研究,指出了酚酸类物质研究中存在的问题,同时展望了酚酸类物质的研究方向与前景。     

Enzymatic synthesis of amino acid-sugar alcohol conjugates in organic media

Amino acid-sugar alcohol conjugates were synthesized by a commercial serine protease, Optimase M-440, in organic media. Optimase M-440 showed broad substrate specificity towards N-t-Boc-protected l-amino acids as acyl donors and sugar alcohols as nucleophiles. Among various solvents tested Optimase M-440 showed the highest activity in pyridine. The regioselective acylation of the primary –OH groups of sugar alcohols gave the amino acid conjugates in good yields without byproducts.     

Enzymatic hydrolysis of hydrophobic triolein by lipase in a mono-phase reaction system containing cyclodextrin; Reaction characteristics

A hydrophobic substrate triolein was hydrolyzed by lipase in a mono-phase reaction system containing cyclodextrin(CD) as emulsifier. The triolein was transformed to an emulsion-like state in the CD containing reaction system in contrast to the oil-droplet like state without CD due to the formation of an inclusion complex between the lipids and CDs. The hydrolysis reaction increased substantially in the CD containing reaction system, and the optimum reaction conditions including the amount of lipase, β-CD concentration, and mixing ratio of triolein and β-CD, were determined. The performance of the enzyme reaction in a mono-phase reaction system was compared with that of a two-phase reaction system which used water immiscible hexane as the organic solvent. The role of a CD in the mono-phase reaction system was elucidated by comparing the degree of the inclusion complex formation with triolein and oleic acid, K_m and V_max values, and product inhibition by oleic acid in aqueous and CD containing reaction systems. The resulting enhanced reaction seems to be caused by two phenomena; the increased accessibility of lipase to triolein and reduced product inhibition by oleic acid through the formation of an inclusion complex.     

Kinetics modeling of the acidolysis with immobilized Rhizomucor miehei lipases for production of structured lipids from sunflower oil

Sunflower oil modification for production of semisolid fats was carried out via acidolysis using palmitic and stearic acids (P + St), hexane and a developed biocatalyst from Rhizomucor miehei lipases. Its kinetic behavior was studied by employing three mathematical models proposed in the literature. Furthermore, a new model was proposed to describe not only the variation of triacylglycerols (TAG), diacylglycerols (DAG), and free fatty acids groups but also the acyl migration reaction occurrence. The effect of the reaction temperature on the kinetic and equilibrium parameters, as well as TAG and reaction intermediates profiles was analyzed. Increasing reaction temperature generated major changes in the overall composition of acylglycerols and gave rise to the highest composition of P + St in the obtained structured lipids (58%, 70 h, 60 °C). P + St incorporation was successfully adjusted by an empirical model (Model I) and a lumped parameter model (Model II) for all the studied reaction times, while the model based on a Ping Pong Bi Bi mechanism (Model III) was only able to describe the kinetics behavior (through the variation of reactant saturated fatty acids concentration) until 24 h. Experimental data were fit satisfactorily by the proposed model (Model IV), showing that the increment in the disaturated TAG formation achieved by the increment in temperature was principally related to the favored DAG formation from triunsaturated TAG.     

Lipase-catalyzed synthesis of phenolic lipids from fish liver oil and dihydrocaffeic acid

The enzymatic synthesis of phenolic lipids by lipase-catalyzed transesterification of dihydrocaffeic acid (DHCA) with fish liver oil was investigated in a selected organic solvent medium. These synthesized phenolic lipids have potential use as nutraceutical products. Using a molar ratio of 1:8 DHCA to fish liver oil in hexane:2-butanone mixtures of 75:25 and 85:15 (v/v), the lipase-catalyzed reaction resulted in maximum conversion of 55.8 and 65.4%, respectively. The maximum conversion of phenolic monoacylglycerols in hexane:2-butanone mixture of 75:25 and 85:15 (v/v) was 40.3 and 37.7%, respectively; using the same solvent mixtures, the conversions of the phenolic diacylglycerol were 15.8 and 36.8%, respectively. Hexane:2-butanone mixture of 75:25 (v/v) was, therefore, the best organic solvent mixture for the production of phenolic monoacylglycerols, while that of 85:15 (v/v) was best for the production of phenolic diacylglycerols. The phenolic lipids produced from the fish liver oil and DHCA demonstrated antioxidant property as indicated by its free radical scavenging capacity.     

Lipase-catalyzed synthesis of phenolic lipids from fish liver oil and dihydrocaffeic acid

The enzymatic synthesis of phenolic lipids by lipase-catalyzed transesterification of dihydrocaffeic acid (DHCA) with fish liver oil was investigated in a selected organic solvent medium. These synthesized phenolic lipids have potential use as nutraceutical products. Using a molar ratio of 1:8 DHCA to fish liver oil in hexane:2-butanone mixtures of 75:25 and 85:15 (v/v), the lipase-catalyzed reaction resulted in maximum conversion of 55.8 and 65.4%, respectively. The maximum conversion of phenolic monoacylglycerols in hexane:2-butanone mixture of 75:25 and 85:15 (v/v) was 40.3 and 37.7%, respectively; using the same solvent mixtures, the conversions of the phenolic diacylglycerol were 15.8 and 36.8%, respectively. Hexane:2-butanone mixture of 75:25 (v/v) was, therefore, the best organic solvent mixture for the production of phenolic monoacylglycerols, while that of 85:15 (v/v) was best for the production of phenolic diacylglycerols. The phenolic lipids produced from the fish liver oil and DHCA demonstrated antioxidant property as indicated by its free radical scavenging capacity.     

Differential incorporation of dietary conjugated linolenic and linoleic acids into milk lipids and liver phospholipids in lactating and suckling rats

Interest in health benefits of conjugated fatty acids is growing. The present study compared the incorporation pattern of dietary conjugated linolenic acids (CLnA) into milk with that of conjugated linoleic acids (CLA). Lactating Sprague-Dawley rats (Day 1) were divided into five groups fed the control diet (n=4) or one of four experimental diets supplemented with 1–2% CLA or CLnA mixture (n=8 each). Supplementation of 1% and 2% CLA led to enrichment of 4.17% and 8.57% CLA, respectively, while supplementation of 1% and 2% CLnA resulted in enrichment of only 0.98% and 1.71% CLnA in the milk lipids, demonstrating the transfer of CLnA from maternal diet to milk was discriminated. When the lactating rats were given a diet containing a CLnA mixture of 9t,11t,13t-, 9c,11t,13t- and 9c,11t,13c-CLnA isomers, two CLA isomers, namely, 9t,11t (0.59–0.90%) and 9c,11t (1.21–1.96%), were found in the milk, suggesting that three CLnA isomers were Δ-13 saturated. Dietary CLnA at 1–2% had no effect on liver phospholipid (PL) fatty acid composition of both maternal and suckling rats, whereas dietary CLA increased docosahexaenoic acid (4c,7c,10c,13c,16c,19c-22:6) and palmitic acid (16:0) proportionally in the PL of maternal rats, but it suppressed 16:0 in the PL of suckling rats. It is concluded that maternal rats incorporate CLnA isomers into milk differently from that of CLA isomers. Most interesting is that maternal rats can metabolically convert CLnA to CLA.     

微生物降解植物化感物质酚酸的研究进展

近年来,农作物连作障碍问题日益严重,给农业生产和可持续发展带来十分不利的影响。连作障碍成因复杂,其中植物化感物质酚酸的毒性作用是重要因素,而消除酚酸最经济、环保、有效的方法是生物防治法。本文综合阐述了利用微生物降解植物化感物质酚酸的菌株资源、酚酸底物范围和降解效果,进而分析论述了微生物降解主要酚酸阿魏酸、香草酸、丁香酸、原儿茶酸等的代谢途径、关键酶及分子机制,推进了对酚酸降解机制的系统认识。在此基础上,阐述了酚酸降解菌在优化作物根际微环境、缓解作物连作障碍及农业资源利用等方面的应用,提出了存在的问题和展望,为深入研究和开发应用微生物资源解决连作障碍等问题、促进现代农业可持续发展提供理论参考。     

Enhancement and inhibition of luminol chemiluminescence by phenolic acids

We explored the behaviour of a series of phenolic acids used as enhancers or inhibitors of luminol chemiluminescence by three different methods to determine if behaviour was associated with phenolic acid structure and redox character. All the phenolic acids inhibited chemiluminescence when hexacyanoferrate(III) was reacted with the phenolic acids before adding luminol. The redox character of these compounds was clearly related to structure. When hexacyanoferrate(III)-luminol-O₂ chemiluminescence was initiated by phenolic acid-luminol mixtures some phenolic acids behaved as enhancers of chemiluminescence, and others as inhibitors. We propose a mechanism to explain these findings. We found direct relationships between the redox character of the phenolic acids and the enhancement or inhibition of the chemiluminescence of the luminol–H₂O₂–peroxidase system and we propose mechanism to explain these phenomena.     

Disbalance of the natural ratio of cations in water as a factor affecting synthesis of lipids and fatty acids in fish eggs

The effects on the lipid status of developing embryos of a disturbed natural ratio of cations in water as a result of the pollution of water bodies by waste with a high potassium content (130–140 mg/l) were studied in the laboratory. The results obtained confirm the indication of reduced lipid synthesis and altered formation of phospholipids in embryos developing in a medium with a disturbed natural ratio of cations. In addition, the lysophospholipid fraction increased in these embryos, which indicates activation of phospholipid hydrolysis. It was also found that changes in the salt regime lead to a decreased content of cholesterol, the main membrane thickener. It was proposed that the changes discovered lead to disturbed stability and permeability of the membranes of fish eggs, with the subsequent death of embryos.     

采煤沉陷区不同造林树种恢复土壤酚酸物质对土壤微生物的影响

酚酸物质是影响微生物群落和结构的最重要因子之一,研究酚酸物质在不同造林树种土壤中的变化规律及其与微生物群落结构的关系,有助于更好地了解和揭示采煤沉陷区恢复造林条件下微生物群落变化的机制.本研究在双鸭山宝山采煤沉陷区的撂荒地基础上营造三针一阔(红松、落叶松、樟子松和杨树)人工林,测定这4种造林地土壤酚类物质、11种酚酸物质和微生物群落结构.结果表明: 复合态酚含量总体表现为人工林显著高于撂荒地,其中,落叶松人工林和杨树人工林的复合态酚含量较高,落叶松人工林和红松人工林的总酚含量显著高于撂荒地,红松人工林的水溶性酚含量显著高于撂荒地;在11种酚酸物质中,阿魏酸、松香酸、β-谷甾醇、齐墩果酸、莽草酸、亚油酸和硬脂酸的含量在人工林土壤中较高.土壤酚类物质与土壤微生物生物量不存在显著的相关关系,个别的酚酸物质与土壤微生物的相关关系显著,其中,阿魏酸、松香酸和β-谷甾醇对土壤微生物生物量有明显的促进作用,与真菌和真菌/细菌存在显著的正相关关系.杨树人工林的酚酸物质含量较高,说明营造杨树人工林对采煤沉陷区的土壤恢复有益.     

Exudation of peroxidase from roots of Festuca rubra and its effects on exuded phenolic acids

Peroxidase is exuded from roots of Festuca rubra under axenic conditions. No phenolase was detected. Peroxidase can use phenolic acids. probably differentially, as hydrogen donors for the H₂O₂ substrate and could thus have an effect on the qualitative and quantitative determinations of phenolic acids also exuded by plants.     

一氧化氮信号在脱落酸诱导丹参酚酸类成分积累中的作用

为探讨一氧化氮(Nitric oxide,NO)信号在脱落酸(Abscisic acid,ABA)诱导丹参酚酸类成分积累中的作用,采用不同浓度一氧化氮外源供体硝普钠(Sodium nitroprusside,SNP)处理丹参毛状根,6 d后采收,测定酚酸类成分含量;ABA联合一氧化氮清除剂(2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide,c-PTIO)或一氧化氮合酶抑制剂(N~G-nitro-L-arginine methyl ester,L-NAME)对丹参毛状根进行处理,测定酚酸类成分含量和关键基因表达量。结果表明,100μmol/L SNP对丹参毛状根中迷迭香酸与丹酚酸B积累的诱导效果最显著,迷迭香酸和丹酚酸B含量分别增加了3倍和4倍。ABA处理能显著促进PAL(Phenylalanine ammonia lyase)、TAT(Tyrosine aminotransferase)和RAS(Rosmarinic acid synthase)基因的表达,促进丹参毛状根中酚酸类成分的积累,而联合c-PTIO或L-NAME共同处理后,3种关键基因表达下调,并显著抑制了酚酸类成分的积累。研究证明NO和ABA均能够促进丹酚酸类成分的积累,NO信号可能介导了ABA对丹酚酸生物合成的诱导作用。