Activity of two lignin biosynthesis enzymes during development of a maize internode

To ascertain the temporal relationships of phcnylpropanoid and lignin pathway enzymes, phenylalanine ammonia-lyase (PAL) and cinnamyl alcohol- NADPH dehydrogenase(CAD), with lignin concentration, PAL and CAD activities and lignin concentrations were assessed during progressive development of the 10th internode in maize (Zea mays L). Enzyme activities were quantified and lignin concentrations were determined by the detergent system of fiber fractiona- tion from lower, middle, and upper sections of the elongating internode harvested 3, 5, 7, 9, 11, 13, and 15 days after the internode had reached 10 mm in length. Over 15 days, a coordinated, sequential, and basipetal pattern of enzyme activities and lignin accumulation evolved through the internode, spanning the stages of rapid elongation, cellular differentiation, and secondary cell wall formation. PAL activity was initiated first, rising to peak activity in elongating tissue, and falling basipetally in tissues as they matured. CAD activity rose in fully elongated, maturing internode tissue where PAL activity was waning following its peak activity. Lignin accumulated in tissues with high CAD activity. CAD activity and lignin deposition patterns were similar: simultaneously increasing in activity and deposition over time while activity and deposition also shifting basipetally through the internode, keeping pace with secondary cell wall formation. Lignin concentration correlated significantly with CAD, but not with PAL.     

用富集培养技术分离具有PAL活性的酵母菌

为获取具有ＰＡＬ活性的菌株，设计了从天然源分离酵母菌的定集培养技术，并从中国西南地区采集２０个样品中得到菌株１４０余株，结果表明本富集培养技术用于从天然源分离具有苯丙氨酸解氨酶活性的菌株是有效、可行的。     

苯丙氨酸及苯丙酮酸对Lactobacillus sp. SK007合成苯乳酸的影响

从自然发酵泡菜中分离筛选到一株乳杆菌(Lactobacillus sp.)SK007,研究了Lactobacillus sp. SK007利用苯丙氨酸合成苯乳酸的过程,结果发现,在MRS培养基中最高可产生0.55 mmol/L苯乳酸,苯丙氨酸剩余94%,但检测不到中间产物苯丙酮酸,这表明苯丙氨酸的转氨反应是苯乳酸合成的限速步骤. 用苯丙酮酸代替苯丙氨酸作为底物可有效突破这一瓶颈,进一步优化了Lactobacillus sp. SK007利用苯丙酮酸合成苯乳酸的发酵条件. 当苯丙酮酸为18.3 mmol/L, 30℃静置培养24 h,苯乳酸产量可达10.25 mmol/L.     

外电场对苯丙氨酸在双水相体系中分配的影响

为减小电极反应和电极距离过大对于两相电泳的影响，设计了微小型多通道两相电泳设备，并以苯丙氨酸为对象开展了双水相电泳的研究工作. 较系统地研究了时间、电流密度、料液初始浓度等因素对葡聚糖–聚乙二醇–水体系中的苯丙氨酸上下相浓度比、萃取率、总传质量以及电流效率的影响. 基于分析，给出了计算最大总传质量的半经验式.     

Phenylalanine Ammonia‐Lyase‐Catalyzed Deamination of an Acyclic Amino Acid: Enzyme Mechanistic Studies Aided by a Novel Microreactor Filled with Magnetic Nanoparticles

Phenylalanine ammonia‐lyase (PAL), found in many organisms, catalyzes the deamination of l ‐phenylalanine (Phe) to (E)‐cinnamate by the aid of its MIO prosthetic group. By using PAL immobilized on magnetic nanoparticles and fixed in a microfluidic reactor with an in‐line UV detector, we demonstrated that PAL can catalyze ammonia elimination from the acyclic propargylglycine (PG) to yield (E)‐pent‐2‐ene‐4‐ynoate. This highlights new opportunities to extend MIO enzymes towards acyclic substrates. As PG is acyclic, its deamination cannot involve a Friedel–Crafts‐type attack at an aromatic ring. The reversibility of the PAL reaction, demonstrated by the ammonia addition to (E)‐pent‐2‐ene‐4‐ynoate yielding enantiopure l ‐PG, contradicts the proposed highly exothermic single‐step mechanism. Computations with the QM/MM models of the N‐MIO intermediates from l ‐PG and l ‐Phe in PAL show similar arrangements within the active site, thus supporting a mechanism via the N‐MIO intermediate.     

Reassessment of treatments to retard browning of fresh‐cut Russet potato with emphasis on controlled atmospheres and low concentrations of bisulphite

The cultivar Pacific Russet with high browning susceptibility was used for most testing. Controlled atmospheres (0.3%, 3% and 21% O₂ in combination with 0%, 6% or 12% CO₂) and anti‐browning chemicals were studied in relation to quality retention and wound‐induced phenolic metabolism of fresh‐cut slices for up to 16 days at 5 °C. The 3% O₂+ 12% CO₂ atmosphere was most effective among those tested, and retarded increases in phenolics and phenylalanine ammonia lyase activity, but had only slight benefit on visual quality. A 1.25% ascorbic acid +1.25% citric acid treatment was ineffective, but when combined with 3% O₂+ 12% CO₂, it was comparable with 0.025% sodium bisulphite. Bisulphite concentrations from 0.05% to 0.25% provided similar effective control of discolouration. Bisulphite as low as 0.025% with 3% O₂+ 12% CO₂ resulted in a visual quality score at the limit of marketability after 8 days at 5 °C. Chemical treatments did not retard increases in phenolic concentrations or phenolic enzyme activities.     

PAL法生产苯丙氨酸的中试研究

报道了苯丙氨酸的扩大试验，在200L种子罐／2500L发醇罐规模上，寻找最佳发醇条件，通过六批培养，转化试验，L－苯丙氨酸平均累积量为30.7g/L.     

Ionizing radiation and marketing simulation on bioactive compounds and quality of grapefruit (Citrus paradisi c.v. Rio Red)

Bioactive compounds in citrus fruits have been shown to be protective against chronic diseases such as cancer and heart disease, but their levels may be affected by postharvest treatments such as storage and irradiation. In this study, grapefruits were exposed to gamma irradiation at 0, 150 and 300 Gy and then stored at 10 °C for 36 d, followed by an additional 20 d at 20 °C. Flavonoid content, terpenoid content, quality (acidity and total soluble solids) and phenylalanine ammonia-lyase (PAL) activity were evaluated at regular intervals during storage. Irradiation and storage affected (P ? 0.05) the levels of bioactive compounds in grapefruit; however, the effect of storage was prominent. Irradiation differentially affected the flavonoid content of pulp and peel. Fruits exposed to 300 Gy had higher (P ? 0.01) narirutin content in peel compared to control fruits at 12 and 56 d after storage. While storage increased the d-limonene and myrcene content in all treatments, control fruit had higher terpenoid content at the end of the storage. PAL activity was found to be in traces in the peel. In general, irradiation or storage had no considerable effect on total soluble solids; however, acidity decreased (P ? 0.05) with storage.     

苯乙烯大孔吸附树脂对苯丙氨酸吸附热力学的研究

测定了D4020苯乙烯大孔吸附树脂对苯丙氨酸的吸附等温线，结果表明Freundlich方程能很好地拟合吸附等温线，吸附过程中随阿斯巴甜母液中NH4Cl含量的增加，D4020树脂对苯丙氨酸的吸附量迅速增加，当NH4Cl浓度大于1．0mol／L时，NH4Cl浓度对吸附量影响很小。当pH〈5．0时，pH对吸附量的影响很大。导出了在pH为5．0时吸附量Q关于温度和NH4Cl离子强度的表达式。     

Fabrication of capped gold nanoparticles by using various amino acids

The production of gold nanoparticles (GNPs) by amino acid is one of the most attractive and interesting subjects in nanobiotechnology. In this study, amino acids have been utilised as a reducing agent and also an agent for capping GNPs. The GNPs were prepared using a reduction solution containing gold cations with optimum concentration of gold salt (5?mM), and also functionalised by glutamic acid, phenylalanine and tryptophan with optimum concentration of amino acids (25?mM). The optimum condition of gold solution and amino acids were achieved by ultraviolet–visible spectroscopy. The size of nanoparticles was obtained 5–20, 10–20 and 20–30?nm, respectively, by transmission electron microscopy and dynamic light scattering techniques. The results obtained from experimental and quantum calculations confirm that amino acids have strong bond while they have anion binding. Moreover, the free carboxylic groups of capped GNPs are one of the suitable and capable beads for binding biological agents. As a result, the medical applications of amino acids and proteins can be used as a practical method due to the strong interaction of peripheral amine groups with nanoparticles.