DyP-Type Peroxidases: Recent Advances and Perspectives

In this review, we chart the major milestones in the research progress on the DyP-type peroxidase family over the past decade. Though mainly distributed among bacteria and fungi, this family actually exhibits more widespread diversity. Advanced tertiary structural analyses have revealed common and different features among members of this family. Notably, the catalytic cycle for the peroxidase activity of DyP-type peroxidases appears to be different from that of other ubiquitous heme peroxidases. DyP-type peroxidases have also been reported to possess activities in addition to peroxidase function, including hydrolase or oxidase activity. They also show various cellular distributions, functioning not only inside cells but also outside of cells. Some are also cargo proteins of encapsulin. Unique, noteworthy functions include a key role in life-cycle switching in Streptomyces and the operation of an iron transport system in Staphylococcus aureus, Bacillus subtilis and Escherichia coli. We also present several probable physiological roles of DyP-type peroxidases that reflect the widespread distribution and function of these enzymes. Lignin degradation is the most common function attributed to DyP-type peroxidases, but their activity is not high compared with that of standard lignin-degrading enzymes. From an environmental standpoint, degradation of natural antifungal anthraquinone compounds is a specific focus of DyP-type peroxidase research. Considered in its totality, the DyP-type peroxidase family offers a rich source of diverse and attractive materials for research scientists.     

DNA barcoding to confirm morphological traits and determine relative abundance of burrowing mayfly species in western Lake Erie

Burrowing mayfly species of the genus Hexagenia are well known indicators of environmental health in lakes and rivers. Two species, H. limbata and H. rigida, are indistinguishable as nymphs and as adult females. Our objectives were to develop a genetic technique to distinguish between the two species and identify morphological features that separate cryptic nymphs and adult females. Fifty nymphs were collected before emergence from 10 sites throughout the western basin of Lake Erie in 2004 and 2005. Using known specimens of adult aerial male H. limbata and H. rigida, we used the mitochondrial cytochrome c oxidase subunit 1 (COI) gene to identify a 16 base pair (bp) difference between species. DNA sequencing confirmed correct species identification based on differences in abdominal pigmentation patterns on adult female imagos in 19 of 20 cases; the lone exception was a female with very faint pigmentation. Pigmentation patterns between species were consistent on nymphs, subimagos and imagos of both sexes. Populations of both species are panmictic across the western basin of Lake Erie, but H. limbata is the numerically dominant species, representing 70 to 100% of nymphs at sites in both years. A separate lineage of H limbata was discovered in the samples. The ability to distinguish nymphs of the two species will aid in developing more sensitive ecosystem indicators.     

Lutein Prevents High Fat Diet‐Induced Atherosclerosis in ApoE‐Deficient Mice by Inhibiting NADPH Oxidase and Increasing PPAR Expression

Epidemiological and experimental studies provide supportive evidence that lutein, a major carotenoid, may act as a chemopreventive agent against atherosclerosis, although the underlying molecular mechanisms are not well understood. The main aim of this study was to investigate the effects of lutein on the alleviation of atherosclerosis and its molecular mechanisms involved in oxidative stress and lipid metabolism. Male apolipoprotein E knockout mice (n = 55) were fed either a normal chow diet or a high fat diet (HFD) supplemented with or without lutein for 24 weeks. The results showed that a HFD induced atherosclerosis formation, lipid metabolism disorders and oxidative stress, but noticeable improvements were observed in the lutein treated group. Additionally, lutein supplementation reversed the decreased protein expression of aortic heme oxygenase‐1 and increased the mRNA and protein expressions of aortic nicotinamide‐adenine dinucleotide phosphate oxidase stimulated by a HFD. Furthermore, the decreased mRNA and protein expression levels of hepatic peroxisome proliferator‐activated receptor‐α, carnitine palmitoyltransferase 1A, acyl CoA oxidase 1, low density lipoprotein receptors and scavenger receptor class B type I observed in mice with atherosclerosis were markedly enhanced after treatment with lutein. Taken together, these data add new evidence supporting the anti‐atherogenic properties of lutein and describing its mechanisms of action in atherosclerosis prevention, including oxidative stress and lipid metabolism improvements.     

Indanones As High‐Potency Reversible Inhibitors of Monoamine Oxidase

Recent reports document that α‐tetralone (3,4‐dihydro‐2H‐naphthalen‐1‐one) is an appropriate scaffold for the design of high‐potency monoamine oxidase (MAO) inhibitors. Based on the structural similarity between α‐tetralone and 1‐indanone, the present study involved synthesis of 34 1‐indanone and related indane derivatives as potential inhibitors of recombinant human MAO‐A and MAO‐B. The results show that C6‐substituted indanones are particularly potent and selective MAO‐B inhibitors, with IC₅₀ values ranging from 0.001 to 0.030 μM . C5‐Substituted indanone and indane derivatives are comparatively weaker MAO‐B inhibitors. Although the 1‐indanone and indane derivatives are selective inhibitors of the MAO‐B isoform, a number of homologues are also potent MAO‐A inhibitors, with three homologues possessing IC₅₀ values <0.1 μM . Dialysis of enzyme–inhibitor mixtures further established a selected 1‐indanone as a reversible MAO inhibitor with a competitive mode of inhibition. It may be concluded that 1‐indanones are promising leads for the design of therapies for neurodegenerative and neuropsychiatric disorders such as Parkinson’s disease and depression.     

P450 BM3 Monooxygenase as an Efficient NAD(P)H‐Oxidase for Regeneration of Nicotinamide Cofactors in ADH‐Catalysed Preparative Scale Biotransformations

Enzymatic oxidations of primary and secondary alcohols catalysed by nicotinamide dependent alcohol dehydrogenases on the preparative scale require cofactor regeneration systems. Of critical value from an economic and ecological perspective is the application of NAD(P)H‐oxidases, which utilise molecular oxygen as a cost‐effective, atom‐efficient and environmentally benign oxidant to regenerate the cofactor NAD(P)⁺. Herein, the P450 BM3 monooxygenase from Bacillus megaterium is presented as an NAD(P)H‐oxidase for the successful regeneration of both NADP⁺ and NAD⁺ on the preparative scale. This enzyme was exemplarily applied for ADH‐catalysed oxidative kinetic resolutions of racemic secondary alcohols and the desymmetrisation of a meso‐diol leading to enantiomerically enriched secondary alcohols in both cases. Furthermore, the ADH‐catalysed oxidation of a primary alcohol targeting the corresponding aldehyde was performed. The obtained results significantly broaden the scope of feasible oxidative biotransformations, thereby increasing the number of synthetic reactions complying with key challenges of a modern and sustainable chemistry such as mild reaction conditions, environmentally benign solvents, and biodegradable non‐toxic catalysts.

     

Fe-chlorophyllin promotes the growth of wheat roots associated with nitric oxide generation

Effects of Fe-chlorophyllin on the growth of wheat root were investigated in this study. We found that Fe-chlorophyllin can promote root growth. The production of nitric oxide in wheat root was detected using DAF-2DA fluorescent emission. The intensity of fluorescent in the presence of 0.1 mg/L Fe-chlorophyllin was near to that observed with the positive control of sodium nitroprusside (SNP), the nitric oxide donor. IAA oxidase activity decreased with all treatments of Fe-chlorophyllin from 0.01 to 10 mg/L. At the relatively lower Fe-chlorophyllin concentration of 0.1 mg/L, the activity of IAA oxidase displayed a remarkable decrease, being 40.1% lower than the control. Meanwhile, Fe-chlorophyllin treatment could increase the activities of reactive oxygen scavenging enzymes, such as superoxide dismutase (SOD) and peroxidase (POD), as determined using non-denaturing polyacrylamide gel electrophoresis. These results indicate that Fe-chlorophyllin contributes to the growth of wheat root associated with nitric oxide generation.     

冷藏对草鱼肉超微结构及代谢物的影响

为解析冷藏草鱼肉品质变化机制,本实验分别采用气相色谱-飞行时间质谱法和透射扫描电子显微镜分析了草鱼肉4 ℃冷藏0、24、72 h后小分子代谢物组成和超微结构变化。结果发现,随着冷藏时间延长,草鱼鱼肉肌纤维间隙变大、排列开始散乱,部分肌节发生断裂。代谢组学共鉴定到211 种代谢物,其中42 种代谢物的含量变化倍数超过1.5 倍（P＜0.05）。乳酸、磷酸累积是鱼肉pH值下降的主要原因。本研究发现三羧酸循环途径代谢物中只有琥珀酸含量显著降低（P＜0.05）,转化成4-羟基丁酸甲酯和γ-氨基丁酸,这3 种化合物水平可能是冷藏草鱼肉新鲜度潜在评价指标。多不饱和脂肪酸（如亚麻酸、花生四烯酸）含量显著下降（P＜0.05）,但壬二酸含量显著增加（P＜0.05）。冷藏过程中,缬氨酸、L-半胱氨酸含量显著增加（P＜0.05）。草鱼肉冷藏过程中,脂肪氧化先于蛋白质降解。草鱼肉中含有的植物次级代谢物（胡椒碱、柚皮素、茉莉酸甲酯、马钱子苷、α-蛔蒿素、枞酸、3,4-二羟基扁桃酸）可通过抑制黄嘌呤氧化酶影响鱼肉核苷酸代谢及鱼肉滋味。     

Peroxidase and polyphenol oxidase thermal inactivation by microwaves in green coconut water simulated solutions

Enzymes from coconut water such as peroxidase (POD) and polyphenol oxidase (PPO) when in contact with oxygen begin reactions causing nutritional and color losses. Solutions simulating the chemical constituents of coconut water were submitted to a batch process in a microwave oven. PPO and POD inactivation data could be characterized by: PPO/water D_93 °C=16.5 s (z=35.5 °C); PPO/sugars D_91 °C=18 s (z=33°C); POD/water D_91.5 °C=44 s (z=24 °C) and POD/sugars D_92 °C=20.5 s (z=19.5 °C). The contact between salts and enzymes promoted a drastic reduction of the initial activity. After the incidence of microwave energy at temperatures above 90 °C, enzymes activity was not detected. These results can indicate an adequate choice of temperature conditions to inactivate coconut water enzymes. The knowledge of how green coconut water constituents influence POD and PPO activity will supply useful information about microwave processing of coconut water.     

邻苯二酚诱导的蓝莓花色苷偶合氧化降解

为了明确蓝莓花色苷在贮藏及加工过程中的降解途径,本文系统地研究了花色苷-多酚氧化酶（Polyphenol Oxidase,PPO）-邻苯二酚的偶合氧化反应机制,探讨了三者之间的量效反应关系。结果表明,邻苯二酚的酶促氧化生成醌类物质和花色苷与醌类物质的氧化反应是偶合氧化体系中主要的两类反应;花色苷的偶合氧化降解符合一级反应动力学模型;当体系中花色苷浓度为54 mg/L时,虽然花色苷的降解速率会随着邻苯二酚浓度的增加而增加,但当邻苯二酚浓度高于1.33 mmol/L时,邻苯二酚浓度的变化对花色苷降解速率影响逐渐变小;花色苷初始浓度越高,邻苯二酚氧化的速率越低,花色苷的降解速率也越低;而酶浓度的增加会导致花色苷的降解速率线性增加。本文结果可推断,在蓝莓贮藏及加工过程中,其组织中的PPO会与其酚类底物结合,产生相应的醌类物质,而这些物质再通过耦合氧化反应导致蓝莓花色苷降解,从而最终导致果实或者相关产品的褐变。且其褐变速率与组织中酶的活性、酚类底物以及花色苷含量都有密切的关系。     

不同杂交后代苹果与富士苹果的多酚氧化酶特性比较

不同杂交后代苹果54号、45号及富士苹果多酚氧化酶特性比较结果表明：不同苹果的最适pH值不同，45号和富士为5．5，54号为6．0；45号、54号和富士最适温度分别为30，35，40℃；富士热稳定性最强，其次是45号，最次为54号；不同苹果PPO的最适底物依次为绿原酸、咖啡酸、儿茶酚；Na2S203对PPO活性抑制效果最佳，L-半胱氨酸与抗坏血酸的抑制效果较好，EDTA-2Na的抑制效果较差；不同苹果PPO同工酶酶带不同，染色深浅不同。富士有四条酶带，45号和54号只有一条共有酶带，富士酶带数最多，染色最深。