π–π Stacking mediated drug–drug interactions in human CYP2E1

Because of having many low molecular mass substrates, CYP2E1 is of particular interests to the pharmaceutical industry. Many evidences showed that this enzyme can adopt multiple substrates to significantly reduce the oxidation rate of the substrates. The detailed mechanism for this observation is still unclear. In the current study, we employed GPU‐accelerated molecular dynamics simulations to study the multiple‐binding mode of human CYP2E1, with an aim of offering a mechanistic explanation for the unexplained multiple‐substrate binding. Our results showed that Thr303 and Phe478 were key factors for the substrate recognition and multiple‐substrate binding. The former can form a significant hydrogen bond to recognize and position the substrate in the productive binding orientation in the active site. The latter acted as a mediator for the substrate communications via π–π stacking interactions. In the multiple‐binding mode, the aforementioned π–π stacking interactions formed by the aromatic rings of both substrates and Phe478 drove the first substrate far away from the catalytic center, orienting in an additional binding position and going against the substrate metabolism. All these findings could give atomic insights into the detailed mechanism for the multiple‐substrate binding in human CYP2E1, providing useful information for the drug metabolism mechanism and personalized use of clinical drugs. Proteins 2013; © 2012 Wiley Periodicals, Inc.     

The reproductive phenology of nine species of Rhodophyta in the subtidal region of the Isle of Man

Monthly samples of about 40 separate plants of each species were collected from 1 to 3 m below lowest astronomical tide on Port Erin breakwater, Isle of Man, Irish Sea. In three species growing on rock, Plocamium cartilagineum, Cryptopleura ramosa and Callophyllis laciniata, about 90% of the plants were fertile in late summer but less than 10% in spring although some fertile plants were always present. Delesseria sanguinea and Odonthalia dentata, also epilithic, had a winter sporing season, Odonthalia extending into late spring, and all plants were sterile in summer. Three species growing epiphytically, Palmaria palmata, Membranoptera alata and Phycodrys rubens, reproduced maximally in the first half of the year at the time when the stipes of the host species, Laminaria hyperborea, grow fastest. Only Palmaria had a sterile season, late summer. The encrusting Cruoria pellita showed little seasonality. The first three species, which reproduce mainly when the sea temperature is above average, are in the northern part of their geographical range. The remaining species (apart from Cruoria) reproduce mainly at low temperatures and are in the southern half of their ranges. Male plants appear to be in a minority in all species, presumably because they were manifest for a shorter period than carposporic plants. They appeared first after sterile periods and were absent as sporing declined. Plocamium and to a lesser extent Cryptopleura show an extremely high preponderance of tetrasporophytes in the population. This is attributed to perennation and some factor disallowing the survival of most of the tetraspores.     

Effects of light and hydrogen peroxide on gene expression of newly identified antioxidant enzymes in the harmful algal bloom species Chattonella marina

ABSTRACT

Antioxidant enzymes are essential proteins that maintain cell proliferation potential by protecting against oxidative stress. They are present in many organisms including harmful algal bloom (HAB) species. We previously identified the antioxidant enzyme 2-Cys peroxiredoxin (PRX) in the raphidophyte Chattonella marina. This enzyme specifically decomposes a hydrogen peroxide (H₂O₂). PRX is the only antioxidant enzyme so far identified in C. marina. This study used mRNA-seq, using Trinity assemble and blastx for annotation, to identify a further five antioxidant enzymes from C. marina: Cu Zn superoxide dismutase (Cu/Zn-SOD), glutathione peroxidase (GPX), catalase (CAT), ascorbate peroxidase (APX) and thioredoxin (TRX). In the gene expression analysis of six enzymes (Cu/Zn-SOD, GPX, CAT, APX, TRX and PRX) using light-acclimated (100 μmol photons m^?2 s^?1) C. marina cells, only PRX gene expression levels were significantly increased by strong light irradiation (1000 μmol photons m^?2 s^?1). H₂O₂ concentration and scavenging activity were also increased and significantly positively correlated with PRX gene expression levels. In dark-acclimated cells, expression levels of all antioxidant enzymes except APX were significantly increased by light irradiation (100 μmol photons m^?2 s^?1). Expression decreased the following day, with the exception of PRX expression. With the exception of CAT, gene expression of antioxidant enzymes was not significantly induced by artificial H₂O₂ treatment, although average gene expression levels were slightly increased in some enzymes. Thus, we suggest that light is the main trigger of gene expression, but the resultant oxidative stress is also a possible factor affecting the gene expression of antioxidant enzymes in C. marina.     

High production of cold-tolerant chitinases on shrimp wastes in bench-top bioreactor by the Antarctic fungus Lecanicillium muscarium CCFEE 5003: Bioprocess optimization and characterization of two main enzymes

The Antarctic fungus Lecanicillium muscarium CCFEE-5003 was preliminary cultivated in shaken flasks to check its chitinase production on rough shrimp and crab wastes. Production on shrimp shells was much higher than that on crab shells (104.6 ± 9.3 and 48.6 ± 3.1 U/L, respectively). For possible industrial applications, bioprocess optimization was studied on shrimp shells in bioreactor using RSM to state best conditions of pH and substrate concentration. Optimization improved the production by 137% (243.6 ± 17.3). Two chitinolytic enzymes (CHI1 and CHI2) were purified and characterized. CHI1 (MW ca. 61 kDa) showed optima at pH 5.5 and 45 °C while CHI2 (MW ca. 25 kDa) optima were at pH 4.5 and 40 °C. Both enzymes maintained high activity levels at 5 °C and were inhibited by Fe⁺⁺, Hg⁺⁺ and Cu⁺⁺. CHI2 was strongly allosamidin-sensitive. Both proteins were N-acetyl-hexosaminidases (E.C. 3.2.1.52) but showed different roles in chitin hydrolysis: CHI1 could be defined as “chitobiase” while CHI2 revealed a main “eso-chitinase” activity.     

Solid-State Fermentation Systems—An Overview

Abstract

Starting with a brief history of solid-state fermentation (SSF), major aspects of SSF are reviewed, which include factors affecting SSF, biomass, fermentors, modeling, industrial microbial enzymes, organic acids, secondary metabolites, and bioremediation. Physico-chemical and environmental factors such as inoculum type, moisture and water activity, pH, temperature, substrate, particle size, aeration and agitation, nutritional factors, and oxygen and carbon dioxide affecting SSF are reviewed. The advantages of SSF over Submerged Fermentation (SmF) are indicated, and the different types of fermentors used in SSF described. The economic feasibilities of adopting SSF technology in the commercial production of industrial enzymes such as amylases, cellulases, xylanase, proteases, phytases, lipases, etc., organic acids such as citric acid and lactic acid, and secondary metabolites such as gibberellic acid, ergot alkaloids, and antibiotics such as penicillin, cyclosporin, cephamycin and tetracyclines are highlighted. The relevance of applying SSF technology in the production of mycotoxins, biofuels, and biocontrol agents is discussed, and the need for adopting SSF technology in bioremediation of toxic compounds, biological detoxication of agro-industrial residues, and biotransformation of agro-products and residues is emphasized.     

Cinnamic acid induced changes in reactive oxygen species scavenging enzymes and protein profile in maize (Zea mays L.) plants grown under salt stress

Plant growth and development are greatly affected due to changes in environmental conditions and become a serious challenge to scientific people. Therefore, present study was conducted to determine the role of secondary metabolites on the growth and development of maize under abiotic stress conditions. Cinnamic acid (CA) is one of the basic phenylpropanoid with antioxidant activity, produced by plants in response to stressful conditions. Response of maize seeds to the presoaking treatment with 0.5 mM CA was studied under different concentrations of NaCl stress. Exogenous CA increased growth characteristics in saline and non-saline conditions, while effects of CA were more significant under saline conditions in comparison to non-saline conditions in maize plants. CA also reduced oxidative damage through the induction of ROS scavenging enzymes such as supperoxide dismutase (SOD) (EC 1.15.1.1), peroxidase (POD) (EC 1.11.1.7), while the activity of enzyme catalase (CAT) (EC 1.11.1.6) was decreased. The content of malondialdehyde (MDA) was reduced significantly in maize leaf under CA treatment. Changes in protein banding patterns in the maize leaves showed a wide variation in response to NaCl-stress, while in the presence of CA salt-induced expression of polypeptides was reduced significantly. Present study clearly reports the alleviative effects of CA in response to salinity stress on growth, metabolic activity and changes in protein profile of 21 days old maize plants.     

不同品种万寿菊对镉胁迫的生长和生理响应

以6个万寿菊品种为研究对象,研究了不同浓度的重金属镉（Cd）胁迫（0、200、400、600 mg/L）对万寿菊基径、株高以及百叶重、茎叶重和单花质量等生长指标及其质膜透性（丙二醛、相对电导率）、保护酶活性（SOD、POD、CAT、APX）以及光合特性(叶绿素含量、净光合速率、气孔导度、蒸腾速率、水分利用效率)等生理指标的影响。结果显示:(1)低于400 mg/L的Cd胁迫对6个万寿菊品种的基径、株高无明显影响,而百叶重、茎叶重和单花质量却随着Cd浓度的增加显著降低,尤其是‘大英雄’和‘迪阿哥’2个品种表现的抗性较差。(2)随着Cd胁迫浓度的增加,各品种丙二醛（MDA）含量和相对电导率（REC）显著增加,而其保护酶活性不同程度降低。(3)Cd胁迫降低了万寿菊的叶绿素Chl a+b含量和Chl a/b,且抗性差的品种降低的幅度更大;叶片净光合速率（Pⁿ）明显降低,但蒸腾速率(T^r)和气孔导度(G^s)相对降幅较小,对水分利用效率（WUE）影响较小。研究表明,不同品种万寿菊对Cd毒害响应存在明显的差异,且Cd胁迫下生理响应的差异是品种间耐性差异的重要原因之一;抗性差的品种在高浓度Cd胁迫下,光合作用能力降低,抗氧化系统酶的活性受到显著抑制,膜质过氧化加剧,且品种‘珍妮’和‘金门’的抗性强于其他品种,可选择作为Cd污染区栽植植物。     

棉花幼苗对不同程度低温逆境的生理响应

以‘新陆早33号'(冷敏感)和‘中棉所50号’(高耐寒)棉花品种为材料,采用人工模拟低温方法,研究不同温度和时间处理下棉花幼苗叶片抗氧化酶活性和渗透调节物质含量的响应特征,探讨不同温度胁迫下棉花的抗逆机制.结果显示:(1)随胁迫温度降低、胁迫时间延长,两参试棉花品种幼苗遭受低温伤害的程度逐渐加剧,尤以5℃和0℃处理表现最为明显.(2)经15℃及10℃处理后,植株叶片REC、POD活性、SP和Pro含量总体上随胁迫时间延长而提高,MDA含量、APX活性和SS含量则先升后降,并均于处理24 h时达峰值;而在5℃及0℃处理下,REC和SOD、POD活性以及MDA、SS、SP含量迅速上升,APX活性、Pro含量则呈先升后降的变化趋势.(3)与‘新陆早33号,相比,‘中棉所50号’在各处理水平下均可保持更大的抗氧化酶活性和渗透调节物质含量上升幅度.研究表明,在不同程度低温逆境中,棉花幼苗可通过调节抗氧化酶活性和渗透调节物质含量,启动相应抗逆调节方式来维持代谢平衡,增强抗逆性;高耐寒性品种在低温逆境中具有更灵敏、高效的活性氧清除能力和渗透调节响应机制,从而有效抵御低温逆境伤害.     

傅家边丘陵山地滨梅根围AM真菌与土壤酶活性的关系

为揭示AM真菌对宿主滨梅（Prunus maritima）的作用特点及对根部土壤酶活性的影响,于2009年4月、7月和10月分别从江苏傅家边丘陵山地滨梅根围分0～10、10～20、20～30、30～40、40～50 cm 5个土层采集土壤样品,观察滨梅AM菌根结构,测定了AM真菌侵染率、孢子密度、土壤磷酸酶、脲酶活性及有效磷、碱解氮含量,着重分析了AM真菌与土壤酶活性之间的关系。结果表明,滨梅能与AM真菌形成良好的共生关系,共生体为泡囊-丛枝结构;AM真菌侵染率和孢子密度分别在7月份和10月份最高,均出现在0～20 cm土层,并随土层加深而下降;AM真菌侵染率与土壤酸性磷酸酶、中性磷酸酶、碱磷酸酶活性显著正相关,而与脲酶活性无相关性;AM真菌孢子密度与碱性磷酸酶、脲酶活性呈极显著正相关关系;孢子密度与土壤有效磷、土壤碱解氮含量显著正相关,但AM真菌侵染率仅与土壤有效磷含量显著正相关;孢子密度与菌根侵染率之间无相关性。可见,滨梅AM真菌侵染率与孢子密度有明显的时空分布并与土壤因子尤其是某些土壤酶活性密切相关,且AM菌根的形成是滨梅适应丘陵山地干旱贫瘠环境的有效对策之一。     

水生生物体内抗氧化酶及其影响因素研究进展

生物体在新陈代谢过程中,细胞内会产生少量的自由基,参与了机体内多种生理过程,具有重要的生理功能。但是在一些应激因子的作用下,体内产生的活性氧大量积累,将会对细胞产生损伤,抗氧化酶类在清除体内活性氧的过程中具有重要作用。在综述了抗氧化酶的分类、结构特点及相互间作用的基础上,阐述了水生生物体内抗氧化酶及其影响因素的研究进展。