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81.

Background

Cyp147G1 is one of 47 cytochrome P450 encoding genes in Mycobacterium marinum M, a pathogenic bacterium with a high degree of sequence similarity to Mycobacterium tuberculosis and Mycobacterium ulcerans. Cyp147G1 is one of only two of these cyp genes which are closely associated with a complete electron transfer system.

Methods

The substrate range of the enzyme was tested in vitro and the activity of CYP147G1 was reconstituted in vivo by co-producing the P450 with the ferredoxin and ferredoxin reductase.

Results

Substrates of CYP147G1 include fatty acids ranging from octanoic to hexadecanoic acid. CYP147G1 catalysed the selective hydroxylation of linear and ω-2 methyl branched fatty acids at the ω-1 position (≥ 98%). Oxidation of ω-1 methyl branched fatty acids generated the ω and ω-1 hydroxylation products in almost equal proportions, indicating altered position of hydrogen abstraction.

Conclusions

This selectivity of fatty acid hydroxylation inferred that linear species must bind in the active site of the enzyme with the terminal methyl group sequestered so that abstraction at the CH bonds of the ω-1 position is favoured. With branched substrates, one of the methyl groups must be close to the compound I oxygen atom and enable hydroxylation at the terminal methyl group to compete with the reaction at the ω-1CH bond.

General significance

Hydroxy fatty acids are widely used for industrial, food and medical purposes. CYP147G1 demonstrates high regioselectivity for hydroxylation at a sub-terminal position on a broad range of linear fatty acids, not seen in other CYP enzymes.  相似文献   
82.
Despite the advancement of the pharmaceutical industry, medicinal plants are still a reliable source of traditional medicines to cure a number of diseases. Various parts of Dillenia pentagyna are used in traditional medicine in India for treatment of various disorders including cancers, but detailed mechanisms are still unknown. Dried leaves of D. pentagyna were extracted with ethanol and termed as an ethanolic extract of leaves of D. pentagyna (EELDP). Our aim was to elucidate the role of EELDP in in-vitro cell migration and apoptosis in highly metastatic human lung adenocarcinoma A549 cells. We measured cell viability and in-vitro cell migration in three different human cancer cells A549, HeLa and U2OS treated with EELDP (0-0.6 mg/mL). However, A549 cells showed higher sensitivity to EELDP treatment. Hence we studied several key markers of metastasis and apoptosis pathway in A549 cells treated with EELDP. EELDP treatment significantly reduced in-vitro cell migration, wound healing, expression and activity of MMP-2, MMP-9 via reduction of nuclear factor kappa Beta (NF-κβ). EELDP also reduced vimentin, N-cadherin and increased claudin-1. The intrinsic pathway of apoptosis was triggered by EELDP via the NF-κβ pathway through the increase of the Bax to Bcl2 ratio, leading to the fall of mitochondrial membrane potential and subsequently induced release of cytochrome c, activation of caspase-3 followed by nuclear fragmentation in A549 cells. Furthermore, we observed change of a few markers of metastasis and apoptosis in other two cell types HeLa and U2OS treated with EELDP. These data implicate that the effect of EELDP is not cell-specific. Since only 0.1 mg/mL EELDP significantly reduces in-vitro cell migration and increases apoptosis, the active compound(s) present in EELDP is very much potent to control highly metastatic cancer.  相似文献   
83.
The Cape Ohma region of Shimokita Peninsula, the northernmost point of Honshu Island, Japan, is subject to both the warm Tsugaru Current and the cold Kurile Current. As a result, the Laminarialean flora includes both cold temperature species (Laminaria japonica Areschoug, Kjellmaniella crassifolia Miyabe and Costaria costata (C. Agardh) Saunders) and warm temperature species (Undaria peterseniana (Kjellman) Okamura, Ecklonia stolonifera Okamura), as well as Undaria pinnatifida (Yendo) Okamura, which is distributed in both waters. The frequency of occurrence (as a measure of distribution) and the biomass of these species were recorded in June 1976 (at 50 points in depths between 8–30 m), July 1988 (192 points, 2.5–25 m) and July 2001 (78 points, 2.5–25 m). Comparison of these data revealed a decrease in cold temperature species and an increase in warm temperature species from 1976 or 1988 to 2001. Long-term data of seawater temperature measured at 5 m depth near the study site showed that mean temperatures in the middle of winter (late January to February) in 1989–2000 were 0.9–1.1 C higher than those in 1980–1988. Higher seawater temperatures in the last decade appear to have affected the frequency of occurrence and biomass of the Laminarialean species along the coasts of Cape Ohma. This result supported our previous conclusion that 1 C higher mean seawater temperature in late January caused a decrease in the biomass of L. japonica (by ca. 64%) along the same coast.  相似文献   
84.
In this investigation, clastogenic effects of Thymus kotschyanus var. glabrescens Boiss. extract (TE) and anticlastogenic effects of this extract against Mitomycin C (MMC) induced chromosome damage have been evaluated in human peripheral blood lymphocytes in vitro. Two series of experiments were conducted. In the first, only 10−5, 10−4, 10−3 and 10−2 μl ml−1 concentrations of TE were used for 48 h to detect potential clastogenicity. In the second, MMC (0.38 μg ml−1) plus 10−5, 10−4, 10−3 and 10−2 μl ml−1 concentrations of TE were used for 48 h to determine anticlastogenic effects. TE did not increase sister chromatid exchanges (SCEs) (except 10−2 μl ml concentration) and chromosome aberrations (CAs) significantly compared with negative and solvent controls. However, it decreased the frequency of MMC induced chromosome aberrations. Decreasing was significant at 10−4, 10−3 and 10−2 μl ml−1 concentrations. On the other hand, TE significantly increased MMC-induced SCEs for all treatment groups compared with positive control.  相似文献   
85.
Genetic responses of the seaweed Ulva pertusa to pine needle ash have been compared using differential display technique. The tissue viability was assessed to evaluate the stress level with triphenyltetrazolium chloride. Total RNA, from tissues treated in seawater containing ash, was reverse transcribed and amplified by PCR with arbitrary primers. The genetic fragments responding to the stress were selectively isolated from agarose gel and sequenced with a DNA auto sequencer. According to sequence analysis, an ash-inducible gene (342 bp) and an ash-suppressed gene (1690 bp) were identified as hypothetical proteins.  相似文献   
86.
The objective of this study was to investigate the influence of interfacial composition and electrical charge on the in vitro digestion of emulsified fats by pancreatic lipase. An electrostatic layer-by-layer deposition technique was used to prepare corn oil-in-water emulsions (3 wt% oil) that contained droplets coated by (1) lecithin, (2) lecithin–chitosan, or (3) lecithin–chitosan–pectin. Pancreatic lipase (1.6 mg mL−1) and/or bile extract (5.0 mg mL−1) were added to each emulsion, and the particle charge, droplet aggregation, and free fatty acids released were measured. In the presence of bile extract, the amount of fatty acids released per unit amount of emulsion was much lower in the emulsions containing droplets coated by lecithin–chitosan (38 ± 16 μmol mL−1) than those containing droplets coated by lecithin (250 ± 70 μmol mL−1) or lecithin–chitosan–pectin (274 ± 80 μmol mL−1). In addition, there was much more extensive droplet aggregation in the lecithin–chitosan emulsion than in the other two emulsions. We postulated that lipase activity was reduced in the lecithin–chitosan emulsion as a result of the formation of a relatively thick cationic layer around each droplet, as well as the formation of large flocs, which restricted the access of the pancreatic lipase to the lipids within the droplets. Our results also suggest that droplets initially coated by a lecithin–chitosan–pectin layer did not inhibit lipase activity, which may have been because the chitosan–pectin desorbed from the droplet surfaces thereby allowing the enzyme to reach the lipids; however, further work is needed to establish this. This information could be used to create food emulsions with low caloric level, or to optimize diets for individuals with lipid digestion problems.  相似文献   
87.
【背景】传统抑菌剂的大量使用导致细菌产生多重耐药性与抗性,而基于细菌群体感应靶点调控的新型抑菌剂可缓解细菌耐药性与抗性,是未来抑菌剂的发展方向之一。【目的】研究连翘(Forsythiasuspensa)提取物对嗜水气单胞菌(Aeromonashydrophila)群体感应系统的影响及可能的作用机制,为新型抑菌剂的开发提供理论依据。【方法】以紫色杆菌(Chromobacterium violaceum)CV026为报告菌株,以嗜水气单胞菌为供试菌株,采用倍比稀释法测定连翘提取物对2种菌的最小抑菌浓度(minimal inhibitory concentration,MIC),通过微量法测定提取物对嗜水气单胞菌生长、群集运动及蛋白酶活性的影响,利用高效液相色谱串联质谱法分析提取物中的主要成分,采用分子对接模拟探究提取物对嗜水气单胞菌群体感应系统的作用机制。【结果】连翘提取物对紫色杆菌CV026和嗜水气单胞菌的MIC均为16.00mg/mL。在亚抑菌浓度下,连翘提取物处理显著抑制了CV026紫色菌素的产生,最大抑制率高达56.30%。经8.00mg/mL连翘提取物处理后,嗜水气单胞菌的群集运...  相似文献   
88.
89.
为了开展油橄榄果渣中主要抗氧化活性成分的研究,该文通过1,1-二苯基-2-苦肼基自由基(DPPH·)法测定了油橄榄果渣不同提取部位清除DPPH·的能力,并采用硅胶、ODS、Sephadex LH-20等柱色谱方法对抗氧化活性部位的化合物进行分离,同时运用核磁共振波谱等方法鉴定了化合物的结构。结果表明:(1)油橄榄果渣的乙酸乙酯部位具有显著的抗氧化活性,其清除DPPH·的半数抑制浓度(IC_(50))为119.11μg·mL^(-1)。(2)从该活性部位分离得到17个化合物,分别鉴定为没食子酸(1)、羟基酪醇(2)、原儿茶酸(3)、酪醇(4)、儿茶素(5)、香草酸(6)、咖啡酸(7)、香草醛(8)、丁香酸(9)、木犀草苷(10)、橄榄苦苷(11)、丁香酚(12)、槲皮素(13)、木犀草素(14)、芦丁(15)、山楂酸(16)、齐墩果酸(17)。其中,化合物1、6、7、9、15为首次从油橄榄果渣中分离得到。该研究明确了油橄榄果渣的抗氧化物质基础,为油橄榄果渣进一步的高值化利用提供了科学依据。  相似文献   
90.
徐来仙  姚兰  周大寨  郭秋菊  朱江  邓楚  艾鑫  夏煜轩 《广西植物》2022,42(11):1949-1958
为探究水杉凋落物的化感作用是否是影响其天然更新的障碍因素,该文设置了8种水杉新鲜凋落物和自然凋落物水浸提液浓度200、100、50、20、10、5、2、1 g·L-1和CK,分析不同类型不同浓度凋落物水浸提液对水杉种子萌发和生长的化感作用的影响。结果表明:(1)水杉新鲜凋落物水浸提液对水杉种子发芽率和发芽势影响不显著(P>0.05),对发芽指数有显著影响(P<0.05),1、100、200 g·L-1显著抑制种子发芽; 水杉自然凋落物水浸提液对水杉种子萌发3个指标影响均不显著(P>0.05),而浓度为5、10、50、200 g·L-1时则有抑制效应。(2)水杉种子芽长、胚轴和主根长度均在不同浓度的新鲜和自然水杉凋落物水浸提液间差异显著(P<0.05),并随着水浸提液浓度的增加(≥10 g·L-1)而逐渐降低,尤其是50~200 g·L-1范围内抑制作用显著增强。(3)新鲜和自然水杉凋落物水浸提液对自身种子萌发后生长的抑制作用相较于对种子萌发强,并且新鲜凋落物的化感效应抑制强度大于自然凋落物。(4)在种子萌发和生长指标中,主根长度对化感物质反应最敏感。(5)水杉种子的芽长生长动态变化符合“S”型生长曲线(R2≥0.988)。总之,水杉凋落物对自身种子萌发和生长具有一定的化感抑制效应,影响其种群天然更新,建议在水杉种群管理中,适当清理林下凋落物,以促进水杉种群的天然更新。  相似文献   
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