P450(BM3) on steroids: the Swiss Army knife P450 enzyme just gets better

The catalytic regio- and stereoselective oxidation of aliphatic C-H bonds by using atmospheric dioxygen as the oxidant is a highly desirable reaction. Reetz and co-workers have recently reported the development of P450(BM3) mutants that oxidise testosterone with greatly increased turnover activity, total stereoselectivity and >95?% regioselectivity.     

高纯度牛血血红素制备工艺研究

以牛血为原料,粗分血球蛋白后用酸性丙酮抽提制备血红素,通过正交实验选取最佳提取条件。实验结果表明,离心时间15min,酸性丙酮、水、氯仿的使用量分别为血液体积的4倍、1倍和5倍,效果较好,血红素纯度达99%以上。     

猪血球水解蛋白的功能特性和抗氧化性

该研究以热处理后的猪血球冻干粉为原料,通过碱性蛋白酶酶解制备水解蛋白质,并对所制备的水解蛋白质的功能特性和体外抗氧化活性进行研究。结果表明,该水解蛋白质具有较好的溶解性、吸湿性和保湿性及一定的起泡性,较弱的乳化性和泡沫稳定性,相对分子量主要分布在2000~3000。体外抗氧化研究表明,水解蛋白质能够有效地清除•OH、DPPH•,抑制脂质过氧化,同时具有较强的总抗氧化能力和金属螯合能力。     

Regulatory FeII/III Heme: The Reconstruction of a Molecule's Biography

More than 20 years of research on heme as a temporary effector molecule of proteins have revealed its widespread impact on virtually all primary functions in the human organism. As our understanding of this influence is still growing, a comprehensive overview of compiled data will give fresh impetus for creativity and developing new strategies in heme‐related research. From known data concerning heme‐regulated proteins and their involvement in the development of diseases, we provide concise information of Fe^II/III heme as a regulator and the availability of “regulatory heme”. The latter is dependent on the balance between free and bound Fe^II/III heme, here termed “hemeostasis”. Imbalance of this system can lead to the development of diseases that were not always attributed to this small molecule. Diseases such as cancer or Alzheimer's disease highlight the reawakened interest in heme, whose function was previously believed to be completely understood.     

Effects of different valences of cerium ion on conformation of Horseradish Peroxidase

Our previous studies demonstrated that Ce^4＋could induce reactive oxygen species （ROS） burst as a signal to promote pacilitaxel biosynthesis in suspension cultured Taxus cuspidate cells. To further understand the mechanism of cerium ions inducing ROS burst, circular dichroism （CD）, synchronous fluorescence, and electron paramagnetic resonance （EPR） were used to detect them inducing conformational change of horseradish peroxidase （HRP）. Horseradish peroxidase activity was reduced by 78% by 0.1 mmol/L Ce^4＋, whereas it was only reduced by 28% by 0.1 mmol/L Ce^3＋. Circular dichroism spectra showed that the percentage of transition from helical content and other structure to β strands and flturns was 23.1 when induced by Ce^4＋, whereas it was only 13.2 when induced by Ce^3＋. In synchronous fluorescence spectra, Ce^4＋ led to red shift and intensity-elevation of tryptophan fluorescence emission maximum, whereas in the case of Ce^3＋, the results were a contrast to the above. Furthermore, g factor （gx and gy） in electron paramagnetic resonance （EPR） induced by Ce^4＋ and Ce^3＋ was significantly different. These results indicated that the different valence of cerium ion induced various conformations of HRP, and Ce^4＋ was more effective than Ce^3＋. This suggested that Ce^4＋ affected the burst of ROS through changing the conformation of oxidoreductase.     

Processing and Frozen Storage Effects on the Iron Content of Cod and Mackerel

Processing and subsequent frozen storage affected the iron content of cod (Gadus morhua) and mackerel (Scomber scombrus) muscle tissue. Frame mince was obtained from the bone rack, without the head or viscera remaining, after filleting. Frame mince had significantly higher iron levels than intact fillets with or without skin or fillets that were subsequently minced. Skin-on fillets had more iron than skin-off fillets. Cod frame mince had about 50% heme iron, while mackerel frame mince ranged from 20-64%. Nonheme iron increased during frozen storage due to heme breakdown. Storage above ?14°C was more deleterious to the heme molecule than lower temperatures (?20°C or ?40°C).     

Heme Burden and Ensuing Mechanisms That Protect the Kidney: Insights from Bench and Bedside

With iron at its core, the tetrapyrrole heme ring is a cardinal prosthetic group made up of many proteins that participate in a wide array of cellular functions and metabolism. Once released, due to its pro-oxidant properties, free heme in sufficient amounts can result in injurious effects to the kidney and other organs. Heme oxygenase-1 (HO-1) has evolved to promptly attend to such injurious potential by facilitating degradation of heme into equimolar amounts of carbon monoxide, iron, and biliverdin. HO-1 induction is a beneficial response to tissue injury in diverse animal models of diseases, including those that affect the kidney. These protective attributes are mainly due to: (i) prompt degradation of heme leading to restraining potential hazardous effects of free heme, and (ii) generation of byproducts that along with induction of ferritin have proven beneficial in a number of pathological conditions. This review will focus on describing clinical aspects of some of the conditions with the unifying end-result of increased heme burden and will discuss the molecular mechanisms that ensue to protect the kidneys.     

Heme Oxygenase-1 Induction by Cobalt Protoporphyrin Ameliorates Cholestatic Liver Disease in a Xenobiotic-Induced Murine Model

Cholestatic liver diseases can progress to end-stage liver disease and reduce patients’ quality of life. Although their underlying mechanisms are still incompletely elucidated, oxidative stress is considered to be a key contributor to these diseases. Heme oxygenase-1 (HO-1) is a cytoprotective enzyme that displays antioxidant action. It has been found that this enzyme plays a protective role against various inflammatory diseases. However, the role of HO-1 in cholestatic liver diseases has not yet been investigated. Here, we examined whether pharmacological induction of HO-1 by cobalt protoporphyrin (CoPP) ameliorates cholestatic liver injury. To this end, a murine model of 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet feeding was used. Administration of CoPP ameliorated liver damage and cholestasis with HO-1 upregulation in DDC diet-fed mice. Induction of HO-1 by CoPP suppressed the DDC diet-induced oxidative stress and hepatocyte apoptosis. In addition, CoPP attenuated cytokine production and inflammatory cell infiltration. Furthermore, deposition of the extracellular matrix and expression of fibrosis-related genes after DDC feeding were also decreased by CoPP. HO-1 induction decreased the number of myofibroblasts and inhibited the transforming growth factor-β pathway. Altogether, these data suggest that the pharmacological induction of HO-1 ameliorates cholestatic liver disease by suppressing oxidative stress, hepatocyte apoptosis, and inflammation.     

血红素铁研究进展

缺铁性贫血是我国人群普遍存在的常见营养性缺乏病,传统的补铁剂存在着吸收率低、副作用大等问题。血红素铁是一种生物态铁,是动物血液中的天然色素和生物铁化合物,能够直接被人体肠粘膜吸收,生物利用率高,而且无副作用,可作为营养强化剂广泛使用,应用于化工、医药和食品行业。本文就血红素铁的功能、应用及其制备、纯化方法等作一综述。     

Altering the regioselectivity of cytochrome P450 CYP102A3 of Bacillus subtilis by using a new versatile assay system

A novel monooxygenase (CYP102A3) has been discovered within the Bacillus subtilis genome that reveals a similarity of 76 % to the well-known cytochrome P450 BM-3 of B. megaterium (CYP102A1). Both enzymes are natural fusion proteins consisting of a heme domain and a FAD/FMN-reductase domain. Because of their high turnover rates, these biocatalysts are of special interest for industrial applications, but show only limited regioselectivity. In this work, the regioselectivity of CYP102A3 was changed by directed evolution and protein design to hydroxylate substrates not only in different subterminal, but also to a high extent, in terminal carbon chain positions. To enable a high-throughput screening procedure, a very versatile assay was developed that is capable of discriminating between terminal and subterminal hydroxylation of carbon chains. A double mutant of CYP102A3 was obtained that produces 48 % octan-1-ol as the main product of the reaction.