过氧化氢驱动的细胞色素P450酶研究进展

细胞色素P450酶在自然界中广泛存在,能催化多种类型的氧化反应,在有机合成和生物化工方面具有重要的应用潜力。尽管大多数P450酶通常需要辅酶和复杂的电子传递体系协助活化氧分子,一些P450酶也可以利用过氧化氢作为末端氧化剂,这极大地简化了催化循环,为P450酶的合成应用提供了一条新的简便途径。本文系统地介绍了几类过氧化氢驱动的P450酶催化体系,包括脂肪酸羟化酶P450SPα和P450BSβ、脂肪酸脱羧酶P450OleTJE、人工改造的羟化酶P450BM3和P450cam突变体、以及基于底物误识别策略的P450-H2O2体系。通过分析催化反应机制,本文探讨了P450-H2O2催化体系在目前存在的挑战和可能的解决途径,并对其进一步应用前景进行了展望。     

Optimized protocol for the pilot-scale preparation of fungal cellulase

Summary A 25-l scale protocol is devised for the optimal secretion and recovery of fungal cellulase. Using a selected higher yieldingTrichoderma viride SMC strain, a protocol consisted of: a) an optimized production medium rich in microcrystalline cellulose (MCC), fortified with 1% (w/v) ammonium sulphate, 0.5% (w/v) soybean flour, 0.1% (v/v) Tween-80 and other trace nutrients; b) optimized physical parameters of production, such as an inoculum containing a homogeneous suspension of 6×10⁷ conidia per 1,28±1°C, pH 4.0±0.5, 300±20 rpm, 11000±1000 l/h aeration, and 170–220 h duration; c) optimal recovery through a filter press (450 l/h rate of filtration) followed by precipitation with 2.5–3.0 volumes of acetone (15°C and basket centrifugation (27°C, 1700 rpm)); and d) vacuum drying (35°C, 4–6 h). This afforded 70% recovery of cellulase in the form of white fluffy powder containing 20000±2000 carboxy methyl cellulase and 1000±50 units filter paperase per g activities, with raw material cost of US$ 8–10 per million carboxy methyl cellulase units. During storage for 18 months at 4°C, ambient temperature and 37°C, the cellulase preparation was found to retain 100, 75 and 60% of its initial activity, respectively.     

Der Einfluß von Rapsextraktionsschrot und Methylthiourazil auf die Schilddrüsensekretionsrate von Hühnern nach kurzfristiger Fütterungsdauer

Three experiments were carried out with male broiler chickens reared from day‐ old to 6 weeks of age on semi‐purified diets containing 10% fresh (Expt. 1 and 3) or oxidized (Expt. 2) re‐esterified triglycerides with a fatty acid composition similar to that of soya bean oil containing increasing concentrations of either a mixture of d‐α‐, γ‐, δ‐tocopherylacetate (d‐tocopherols) of natural source or dl‐α‐ tocopheryl acetate (dl‐tocopherol). In Expt. 1 and 2 the mixture of d‐tocopherols consisted of 35.7% d‐α‐, 45.3% d‐γ‐ and 19.0% d‐δ‐, while in Expt. 3 the distribution was 25.3% d‐α‐, 28.1% d‐γ‐and 10.8% d‐γ‐ in 35.8% re‐ esterified triglycerides. The relative biopotency of d‐α‐: γ‐: δ‐tocopherol was anticipated to be 100: 25: 1, whereas that of dl‐a‐tocopherol was 74% relative to d‐ a‐tocopherol. The experiments demonstrate that the results obtained for the biological activity depend on the response parameters chosen. With respect to gain in weight, feed conversion, relative organ weight, packed cell volume (PCV), ELP (erythrocytelipidperoxidation), plasma activities of glutamate‐oxaloacetate‐transaminase (GOT), creatine kinase (CK) and glutathione peroxidase (GSH‐Px) and plasma Na⁺ concentration, the mixture of natural source tocopherols was identical to that of dl‐α‐tocopheryl acetate, although the concentration of a‐ tocopherol was only about one third of that of dl‐α‐tocopherol. Differences between natural source and synthetic tocopherols were expectedly observed with respect to plasma concentrations of α‐, γ‐, δ‐tocopherol. Differences between the two forms as to muscular dystrophy, in vitro haemolysis and potassium concentration in plasma were ambigous. It is suggested that the function of d‐α‐, γ‐, δ‐tocopherol in erythrocyte fragility and skeletal muscle structure should be compared to that of dl‐α‐tocopherol in future investigations.     

近地面激光雷达点云密度对森林冠层结构参数提取准确性的影响

基于激光雷达技术获取冠层结构为森林生态学研究增加了新的维度。搭载于多旋翼无人机的近地面激光雷达相比于固定翼有人机的机载激光雷达,能够更加灵活高效地获取森林群落样地高密度点云。但在实际操作中,往往出现局部低密度点云数据,影响了冠层结构参数提取的准确性。使用4块森林动态样地的近地面激光雷达点云数据;利用航带分解方法分析各样地低密度样方成因;采用点云抽稀模拟算法计算并拟合偏差曲线,对比不同样地、参数和取样尺度间的点云密度对冠层结构参数提取准确性的影响;根据偏差曲线计算各条件下保证参数提取准确性的最低点云密度。结果发现：1)低密度区域主要受地形或(和)近地面遥感设计规划的影响。地形复杂的测区(西双版纳和古田山样地),遥感规划难度大,整体难以获取高密度点云(在30点/m²左右),容易在沟谷和高海拔处出现低密度样方。平坦测区(长白山两块样地)虽可获取高密度点云(均超过150点/m²),但欠佳的遥感规划设计导致长白山1测区北部出现1hm²低密度区域。2)冠层参数提取准确性随点云密度减少而迅速降低,呈负指数幂函数关系。这一变化趋势在不同...     

Detecting Resistance to Organophosphates and Carbamates in the Cattle Tick Boophilus Microplus, with a Propoxur-based Biochemical Test

Rapid and sensitive detection of resistance to insecticides in arthropods is needed. In the cattle tick, Boophilus microplus, resistance to a variety of acaricides is widespread. The most commonly used assay for resistance, the larval packet test, takes at least two, but generally six weeks for a one-host tick like B. microplus to complete and may take up to three months to complete for three-host ticks. Here we describe a test for resistance to organophosphate acaricides that can be used on larvae and adult ticks which takes less than 24 hours. The test measures the difference in acetylcholinesterase (AChE) activity in homogenates of ticks in the presence and absence of propoxur, a carbamate acaricide. We found clear discrimination of organophosphate-susceptible and organophosphate-resistant adults with 100 M propoxur. AChE from susceptible ticks had almost no activity at this concentration of propoxur whereas AChE from resistant ticks had 67% of its potential activity. AChE from heterozygote ticks could also be distinguished from AChE from homozygous-susceptible and homozygous-resistant ticks. This is the first biochemical test for resistance to an acaricide. Rapid, sensitive tests like ours will allow resistance to organophosphates to be detected soon after it develops in the field, thus, the spread of resistance might be slowed and the useful life of acaricides extended.     

Offspring performance and female preference of Pagiophloeus tsushimanus (coleoptera: curculionidae) on three Lauraceae tree species: A potential risk of host shift caused by larval experience

The weevil Pagiophloeus tsushimanus Morimoto (Coleoptera: Curculionidae), native to Eastern Asia, is a wood-boring pest that causes severe damage to camphor trees (Cinnamomum sp.) in Shanghai, China. Other Lauraceae tree species that grew sympatrically with this pest in close proximity could face a potential threat. To assess the potential risks of host shift, we explored the phenotypic associations between preference and performance in P. tsushimanus reared on three Lauraceae tree species. In a no-choice experiment offering branches of each plant as diet material and oviposition sites, we found that individuals reared on Cinnamomum camphora (L.) Presl (Laurales: Lauraceae) exhibited the strongest performance with shorter development time, higher survival and growth rate in the immature stage, longer longevity and greater fecundity in adults. In contrast, those on novel Lauraceae tree species (Cinnamomum chekiangensis Nakai and Phoebe chekiangensis Shang) had difficulty completing their whole life cycle due to significantly lower survival and reproduction. In a multiple-choice experiment, C. camphora was established as the preferred host. However, we found that the larval experiences on the non-preferred host plants contributed to an increased preference for that plant species. These results indicated that both the preference-performance hypothesis and the Hopkins’ host selection principle are applicable in this weevil under experimental conditions. It is possible that although the weevil performed poorly on two novel Lauraceae tree species, under favourable conditions their surviving offspring could evolve into a new host-specific population. Consequently, this weevil pest needs to be monitored on these novel Lauraceae tree species.     

Foliar Application of Phosphorus Enhances Photosynthesis and Biochemical Characteristics of Maize under Drought Stress

Water is essential for the growth period of crops; however, water unavailability badly affects the growth and physiological attributes of crops, which considerably reduced the yield and yield components in crops. Therefore, a pot experiment was conducted to investigate the effect of foliar phosphorus (P) on morphological, gas exchange, biochemical traits, and phosphorus use efficiency (PUE) of maize (Zea mays L.) hybrids grown under normal as well as water deficit situations at the Department of Agronomy, University of Agriculture Faisalabad, Pakistan in 2014. Two different treatments (control and P @ 8 kg ha⁻¹ ) and four hybrids (Hycorn, 31P41, 65625, and 32B33) of maize were tested by using a randomized complete block design (RCBD) with three replications. Results showed that the water stress caused a remarkable decline in total soluble protein (9.7%), photosynthetic rate (9.4%) and transpiration rate (13.4%), stomatal conductance (10.2%), and internal CO₂ rate (20.4%) comparative to well-watered control. An increase of 37.1%, 36.8%, and 24.5% were recorded for proline, total soluble sugar, and total free amino acid, respectively. However, foliar P application minimized the negative impact of drought by improving plant growth, physio-biochemical attributes, and PUE in maize plants under water stress conditions. Among the hybrids tested, the hybrid 6525 performed better both under stress and non-stress conditions. These outcomes confirmed that the exogenous application of P improved drought stress tolerance by modulating growth, physio-biochemical attributes, and PUE of maize hybrids.     

Depolymerised carrageenan enhances physiological activities and menthol production in Mentha arvensis L.

Irradiated carrageenan (IC) could elicit plant growth promoting activities in plants. The effect of foliar spray of five concentrations of IC (20, 40, 60_, 80 and 100 mg L⁻¹) was studied on Mentha arvensis L. in terms of plant growth, physiological attributes, herbage yield and the content and yield of essential oil and its components. Un-irradiated carrageenan and deionized water had no effect on the attributes studied. GPC study revealed formation of low molecular weight fractions in irradiated samples containing less than 20,000 molecular weight oligomers which are responsible for plant growth promotion in this study. 80 mg L⁻¹ of IC was the most effective concentration which resulted in the highest values of growth attributes, herbage yield and the content and yield of essential oil and menthol content of the oil. It also improved the leaf-nutrient contents, photosynthetic rate and other physiological parameters. 100 mg L⁻¹ of IC did not further improve the attributes studied, but it was always better than the control.     

Effects of four constant temperatures on the development of two Bradysia (Diptera: Sciaridae) species

In this study, the effects of temperature on the growth, development, survival, fecundity and other population parameters of two local Bradysia species B. odoriphaga and B. impatiens were studied at four constant temperatures (25, 28, 31 and 34°C). The results show that 25°C is the optimum temperature for the growth and development of B. odoriphaga, while 28°C is more favourable for B. impatiens. The temperature of 31°C restricted the growth and development, while the temperature of 34°C inhibited the eggs hatching in both species, resulting in no egg survival and no subsequent development. High temperatures (>28°C) prolonged the 4th larval stage duration, mean generation time (T) and population doubling time (Dt) of both species. The high temperature of 31°C greatly shortened the female longevity, weakened the oviposition and reduced the survival of both species. Moreover, the life table parameters R₀, r_m and λ were also suppressed by this high temperature. However, the high temperature of 31°C had little impact on the egg survival, pupal weight and male longevity. In addition, at 31°C, the values of R₀, r_m and λ of B. odoriphaga were higher than those of B. impatiens, suggesting that B. odoriphaga is more tolerant to high temperature than B. impatiens. The differences between two Bradydsia species seem determined genetically. Our findings are important for better understanding their biological characteristics at a certain constant temperature and demonstrate the possibility to control and manage those two Bradysia species by increasing ambient temperature.