碱蓬籽油脂的超临界CO_2提取工艺优化

碱蓬籽中油脂含量丰富,尤以不饱和脂肪酸甘油酯-亚油酸酯居多,使用超临界CO2提取碱蓬籽油,确定最佳的提取条件。以亚油酸酯得率为指标,考察了提取温度、提取压力、提取时间、夹带剂流量对实验的影响,在单因素的基础上,采用响应面法对工艺进行优化,得到的最佳提取条件为:提取温度46℃,提取压力23.5MPa,提取时间137min,夹带剂流量0.04m L·min-1,在此条件下亚油酸酯得率为15.29%。优化的提取工艺可行,对工业化生产有一定的指导意义。     

Structural modeling and docking studies of ribose 5-phosphate isomerase from Leishmania major and Homo sapiens: A comparative analysis for Leishmaniasis treatment

Leishmaniases are caused by protozoa of the genus Leishmania and are considered the second-highest cause of death worldwide by parasitic infection. The drugs available for treatment in humans are becoming ineffective mainly due to parasite resistance; therefore, it is extremely important to develop a new chemotherapy against these parasites. A crucial aspect of drug design development is the identification and characterization of novel molecular targets. In this work, through an in silico comparative analysis between the genomes of Leishmania major and Homo sapiens, the enzyme ribose 5-phosphate isomerase (R5PI) was indicated as a promising molecular target. R5PI is an important enzyme that acts in the pentose phosphate pathway and catalyzes the interconversion of d-ribose-5-phosphate (R5P) and d-ribulose-5-phosphate (5RP). R5PI activity is found in two analogous groups of enzymes called RpiA (found in H. sapiens) and RpiB (found in L. major). Here, we present the first report of the three-dimensional (3D) structures and active sites of RpiB from L. major (LmRpiB) and RpiA from H. sapiens (HsRpiA). Three-dimensional models were constructed by applying a hybrid methodology that combines comparative and ab initio modeling techniques, and the active site was characterized based on docking studies of the substrates R5P (furanose and ring-opened forms) and 5RP. Our comparative analyses show that these proteins are structural analogs and that distinct residues participate in the interconversion of R5P and 5RP. We propose two distinct reaction mechanisms for the reversible isomerization of R5P to 5RP, which is catalyzed by LmRpiB and HsRpiA. We expect that the present results will be important in guiding future molecular modeling studies to develop new drugs that are specially designed to inhibit the parasitic form of the enzyme without significant effects on the human analog.     

Noncanonical Reactions of Flavoenzymes

Enzymes containing flavin cofactors are predominantly involved in redox reactions in numerous cellular processes where the protein environment modulates the chemical reactivity of the flavin to either transfer one or two electrons. Some flavoenzymes catalyze reactions with no net redox change. In these reactions, the protein environment modulates the reactivity of the flavin to perform novel chemistries. Recent mechanistic and structural data supporting novel flavin functionalities in reactions catalyzed by chorismate synthase, type II isopentenyl diphosphate isomerase, UDP-galactopyranose mutase, and alkyl-dihydroxyacetonephosphate synthase are presented in this review. In these enzymes, the flavin plays either a direct role in acid/base reactions or as a nucleophile or electrophile. In addition, the flavin cofactor is proposed to function as a “molecular scaffold” in the formation of UDP-galactofuranose and alkyl-dihydroxyacetonephosphate by forming a covalent adduct with reaction intermediates.     

The Effects Of Humic Substances On Pinus Callus Are Reversed By 2,4-Dichlorophenoxyacetic Acid

The reversal of humic matter-induced inhibition of callus growth and metabolism by 2,4-dichlorophenoxyacetic acid (2,4-D) was studied in Pinus laricio. Two forest humic fractions (relative molecular mass (M_r) > 3500), derived from soil under Fagus sylvatica (Fs) and Abies alba (Aa) plantation, were used. Pinus laricio callus was grown for a subculture period (4 weeks) on Basal Murashige and Skoog (MS) medium plus forest humic matters (Fs or Aa), at a concentration of 1 mg C/l, and then was transferred, for an additional four weeks, to a MS medium culture without humic matter, but with different hormones: indole-3-acetic acid (IAA, 2 mg/1) or 2,4-dichlorophenoxyacetic acid (2,4-D, 0.5 mg/1) and/or 6-benzylaminopurine (BAP, 0.25 mg/1). Growth of calluse, glucose, fructose, and sucrose contents, and activities of soluble and bound invertases, glucokinase, phosphoglucose isomerase, aldolase, and pyruvate kinase were monitored. The results show a negative effect of humic fractions on callus growth, due to decreased utilization of glucose and fructose, and decreased activities of glycolytic enzymes. The effects are reversible. Substitution of humic fractions with 2,4-D+BAP or 2,4-D is followed by an increase of glycolytic enzyme activities and, consequently, by the utilization of glucose and fructose that induces a restart of growth. In contrast, the inhibitory effects of humic fractions persist when they are substituted with BAP alone, indicating that only the auxin 2,4-D is capable of reversing the negative effects. A possible competitive action on the auxin-binding site between 2,4-D and the chemical structures in the forest humic fractions is suggested.     

Theoretical examination of the mechanism of aldose-ketose isomerization

Two mechanisms for an aldose–ketose isomerization havebeen examined using high level ab initio and semiempirical molecularorbital methods. The proton transfer pathway via an enediolintermediate is shown to be favored in the absence of a metalion, while the hydride transfer pathway becomes favored in thepresence of a metal ion. Our calculations explain why the protontransfer pathway is operative in most aldose–ketose isomerizationreactions. These calculations also provide further support forthe previously proposed metal ion-mediated hydride transfermechanism for xylose isomerase.     

An evaluation of commercial nickel catalysts during hydrogenation of soybean oil

The activities of several commercial nickel catalysts were determined by measuring their activation energies. Among these catalysts, G95E, Resan 22, Nysosel 222 and 325, all with low activation energy, were more active than DM3 and G95H, which had higher activation energy. However, the less active catalysts increased the linoleate selectivity of soybean oil during hydrogenation. The yields of bothtrans isomers and winterized oil were higher for the more selectively hydrogenated oil catalyzed by the less active catalysts. In the sensory evaluation, the fractionated solid fat that contained moretrans isomers was lower in flavor scores than the fractionated liquid oil after hydrogenation and winterization of soybean oil.     

Antioxidant efficacy of amino acids in methyl linoleate at different relative humidities

Antioxidant efficacy of the amino acids methionine, tryptophan, aspartic acid, serine, alanine and arginine in methyl linoleate were compared to a methyl linoleate control at 2,50 or 79% relative humidity (RH) at 37°C. Antioxidant efficacy varied with RH and the individual amino acids. Arginine had the highest antioxidant efficacy at all RH values compared to the control. The efficacy of alanine was equal to that of arginine at RHs of 50 and 79% but was lower at 2% RH. The presence of aliphatic, alkaline amino, hydroxyl or thiol groups in the side chain of the amino acids increased the antioxidant efficacy at high RHs.     

嗜酸乳杆菌亚油酸异构酶的结构与催化机理

对嗜酸乳杆菌亚油酸异构酶基因及其启动子进行了定位,采用多种软件分析了酶蛋白中α螺旋、β折叠等二级结构的组成以及酶蛋白中的糖基化位点、磷酸化位点的分布.证明了该蛋白具有肌凝蛋白交叉反应抗原、链状球菌的67 000肌凝蛋白交叉反应抗原、番茄红素脱氢酶相关蛋白和FAD结合蛋白等4个功能结构域,并首次提出其催化共轭亚油酸合成时可能存在氧化还原反应过程.     

海藻酸钠固定化亚油酸异构酶及其酶学性质初步研究

以海藻酸钠为载体固定化亚油酸异构酶。研究了海藻酸钠浓度、酶用量、CaC12浓度、固定化时间对固定化过程的影响。结果表明,最佳固定化工艺是：以4%的海藻酸钠为载体、应用海藻酸钠溶液用量与酶液量的体积比3：1、3%的CaC12固定化5h。固定化酶的最适pH值为7.0,与游离酶相比,提高了0.5个pH单位;固定化酶和游离酶最适温度分别为35℃和30℃;固定化酶比游离酶具有更好的温度和pH值适应性。     

Effect of processing on the antioxidant activity of millet grains

Millets are generally dehulled and subjected to a hydrothermal treatment before consumption, thus the hulls can be used as a potential source of antioxidants. Several millet grains, namely kodo, finger (Ravi), finger (local), proso, foxtail, little and pearl millet were studied. Antioxidant activities of phenolic extracts obtained from whole grains, as well as their corresponding dehulled and cooked grains and hulls were studied for their total phenolic content (TPC), radical scavenging capacity, and antioxidant activity in a β-carotene/linoleate emulsion. The phenolics present in whole grains were identified and quantified using HPLC and HPLC/MS and results were expressed as total for each of the phenolic groups. The TPC ranged from 2 to 112 μmol ferulic acid equivalents/g defatted meal. All varieties exhibited effective inhibition of 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl, peroxyl and superoxide radicals. Dehulling and cooking affected the TPC and radical scavenging and antioxidant activities of the grains, depending on the variety. In general, the antioxidant activity of phenolic extracts was in the order of hull > whole grain > dehulled grain > cooked dehulled grain With the exception of the two finger millet varieties, hulls of other millet grains had high TPC, thus demonstrating their superior antioxidant activity. Hydroxybenzoic acids, hydroxycinnamic acids and flavonoids in whole grains were identified as contributors to the observed effects. Therefore, dehulling of grain and hydrothermal treatments affect the phenolic content and antioxidant potential of millet grains.