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1.
《Bioscience, biotechnology, and biochemistry》2013,77(5):1112-1118
During previous work on deriving inosine-producing mutants of Escherichia coli, we observed that an excess of adenine added to the culture medium was quickly converted to hypoxanthine. This phenomenon was still apparent after disruption of the known adenosine deaminase gene (add) on the E. coli chromosome, suggesting that, like Bacillus subtilis, E. coli has an adenine deaminase. As the yicP gene of E. coli shares about 35% identity with the B. subtilis adenine deaminase gene (ade), we cloned yicP from the E. coli genome and developed a strain that overexpressed its product. The enzyme was purified from a cell extract of E. coli harboring a plasmid containing the cloned yicP gene, and had significant adenine deaminase [EC 3.5.4.2] activity. It was deduced to be a homodimer, each subunit having a molecular mass of 60 kDa. The enzyme required manganese ions as a cofactor, and adenine was its only substrate. Its optimum pH was 6.5-7.0 and its optimum temperature was 60°C. The apparent Km for adenine was 0.8 mM. 相似文献
2.
Ahmed H. Moustafa Said A. Said Abd-El Fattah Z. Haikal Rajab Abu-El-Halawa Rimaa T. Abd- El kader 《Nucleosides, nucleotides & nucleic acids》2013,32(3):111-128
Hydroquinoline acyclonucleosides 2, 4, 6a,b, 8a,b, 9a,b, and their corresponding N-alkyl derivatives (10–12) were obtained by the reaction of 1a,b with acetoxybutylbromide, (2-acetoxyethoxy)methyl bromide, 3-chloropropanol, 1,3-dichloro-2-propanol, epichlorohydrin, propargyl/allyl bromides in the presence of K2CO3 in dry dimethylformamide (DMF). In a similar manner, reaction of 1a,b with glycosyl/galactosyl and lactosyl bromide afforded the corresponding N-nucloside derivatives 13a,b, 15a,b, and 17, respectively. Deacetylation of the N-nucleosides derivatives in the presence of Et3N/MeOH and few drops of water gave the deprotected derivatives 3, 5, 7a,b, 14a,b, 16a,b, and 18 in good yields, respectively. All the newly synthesized compounds are elucidated by infrared, 1H, 13C NMR and elemental analyses. Some of these compounds were screened for antimicrobial activities. 相似文献
3.
《Bioscience, biotechnology, and biochemistry》2013,77(10):2322-2325
An 11-residue peptide lactone, termed the gelatinase biosynthesis-activating pheromone (GBAP), triggers the production of the pathogenicity-related extracellular proteases, gelatinase and serine protease, in Enterococcus faecalis. In this study, we synthesized GBAP and its analogs and examined their gelatinase biosynthesis-inducing activity. This study on the structure-activity relationship shows that a lactone ring was indispensable for the activity. 相似文献
4.
Khomutov M. A. Mikhura I. V. Kochetkov S. N. Khomutov A. R. 《Russian Journal of Bioorganic Chemistry》2019,45(6):463-487
Russian Journal of Bioorganic Chemistry - Biogenic polyamines spermine and spermidine are present in eukaryotic cells in micro- and millimolar concentrations, that determines the multiplicity of... 相似文献
5.
RecQ家族解旋酶是DNA解旋酶中高度保守的一个重要家族,在维持染色体的稳定性中起着重要的作用.人类RecQ家族解旋酶突变会导致几种与癌症有关的疾病.本研究旨在诱导大肠杆菌RecQ解旋酶体外表达,并应用生物化学和生物物理学技术研究大肠杆菌RecQ解旋酶的生物学活性. 体外诱导表达获得纯度达90% 以上并具有高活性的大肠杆菌重组RecQ解旋酶,其可溶性好;经生物学活性分析显示具有DNA结合活性、ATP依赖的DNA解链活性、DNA依赖的ATP酶活性. 较之双链DNA(dsDNA),大肠杆菌RecQ解旋酶更容易与单链DNA(ssDNA)结合( P<0.01 ),但与长度不同的dsDNA的结合特性有差异(P<0.01)而与ssDNA没有差异(P>0.05);大肠杆菌RecQ解旋酶对3种dsDNA的解链速率不同(P<0.05);大肠杆菌RecQ解旋酶的ATP酶活性与辅助因子ssDNA长度也呈正相关(P<0.01). 这些研究结果将有助于阐明大肠杆菌RecQ解旋酶的分子作用机制,并为研究RecQ解旋酶家族其它成员的结构与功能提供帮助. 相似文献
6.
Naoshi Nakagawa Karl J. Kramer Kenji Mori 《Bioscience, biotechnology, and biochemistry》2013,77(10):2381-2382
Encapsulated calcium in liposome (L-Ca) produced by using egg phosphatidyl choline in the laboratory was injected into rabbit to evaluate the effect of calcium injection on the ageing of meat. After injecting L-Ca into the blood vessels of rabbit to increase the Ca2+ concentration in the body for 24 h, the fragmentation rate of myofibrils was observed. The fragmentation rates in the loin from the control group and L-Ca injected group were 2.56% and 3.10% in 2 days, 12.27% and 16.18% in 6 days, and 33.56% and 49.60% in 10 days, respectively (p<0.05). SDS–PAGE patterns of connectin and nebulin show that the total degradation of connectin by the control group took longer than 2–3 days, while it was within 1 day for the L-Ca-injected group. The control group took 8–10 days for nebulin, while the L-Ca-injected group took 2–3 days for total degradation. These results indicate that, injecting L-Ca into rabbit was effective for reducing the ageing period of meat without resulting in any physical shock or contamination. 相似文献
7.
Shih Hsi Chu Zhi Hao Chen Elizabeth C. Rowe Fardos N. M. Naguib Hahmoud H. el Kouni Ming Y. Chu 《Nucleosides, nucleotides & nucleic acids》2013,32(3):303-311
Abstract Hydroxymethyl analogs of 5-benzylacyclouridine (BAU) and 5-benzyloxybenzylacyclouridine (BBAU) were synthesized by the condensation of appropriately blocked 2-(chloromethyl)glycerols with substituted 2, 4-dimethoxypyrimidines. The HM derivatives were found to be potent inhibitors of the enzyme uridine phosphorylase and to potentiate significantly the growth-inhibiting action of FdUrd in cell culture. 相似文献
8.
《Bioscience, biotechnology, and biochemistry》2013,77(11):2992-2997
Analogs of cortistatins, a series of anti-angiogenic compounds isolated from the Indonesian marine sponge Cortisium simplex, were synthesized from estrone by using the Suzuki-Miyaura coupling reaction as the key step. The estrone-isoquinoline hybridized compound showed selective inhibitory activity against the proliferation and VEGF-induced migration of HUVEC. 相似文献
9.
Arabidopsis nucleoside hydrolases involved in intracellular and extracellular degradation of purines
Recently, the first plant nucleoside hydrolase, NSH1 (former designation URH1), was identified at the molecular level. This enzyme's highest hydrolysis capacity is for uridine, thereby balancing pyrimidine salvage and catabolism. NSH1 was found to be less efficient in the hydrolysis of further nucleosides. However, it remained unclear whether purine nucleosides are processed by NSH1. Moreover, the biochemical and physiological functions of further NSH isoforms in Arabidopsis has not been analyzed. Here we show that NSH1 is also able to hydrolyze xanthosine with high efficiency, and thus represents the leading activity in purine and pyrimidine breakdown in a cell. A knockout mutant for NSH1 showed symptoms of accelerated senescence, accompanied by marked accumulation of uridine and xanthosine under conditions of prolonged darkness. The closest, so far uncharacterized, homolog of NSH1, NSH2, was found to act during the late phase of senescence and may support inosine breakdown. NSH3, another NSH isoform, surprisingly functions as an extracellular, purine-specific hydrolase that is involved in degradation of extracellular nucleosides and may participate in wound and pathogen responses. 相似文献
10.
Toshio Yanaki Wataru Itoh Kengo Tabata 《Bioscience, biotechnology, and biochemistry》2013,77(9):2415-2416
trans-3-Methylthioacrylamide (3-MTAA-NH2) was isolated as colorless needles from the culture broth of Streptomyces sioyaensis, a siomycin-producer. This substance is considered to be not only a new metabolite from methionine but also a new substance. The isolation and identification of 3-MTAA-NH2, as well as the cultural conditions for production, were investigated. A variety of other Streptomyces also produced 3-MTAA-NH2 from methionine. 相似文献
11.
Julia Schollmeyer Saskia Waldburger Kendra Njo Heba Yehia Anke Kurreck Peter Neubauer Sebastian L. Riedel 《Biotechnology and bioengineering》2023,120(11):3322-3334
Nucleoside phosphorylases are important biocatalysts for the chemo-enzymatic synthesis of nucleosides and their analogs which are, among others, used for the treatment of viral infections or cancer. S-methyl-5′-thioadenosine phosphorylases (MTAP) are a group of nucleoside phosphorylases and the thermostable MTAP of Aeropyrum pernix (ApMTAP) was described to accept a wide range of modified nucleosides as substrates. Therefore, it is an interesting biocatalyst for the synthesis of nucleoside analogs for industrial and therapeutic applications. To date, thermostable nucleoside phosphorylases were produced in shake flask cultivations using complex media. The drawback of this approach is low volumetric protein yields which hamper the wide-spread application of the thermostable nucleoside phosphorylases in large scale. High cell density (HCD) cultivations allow the production of recombinant proteins with high volumetric yields, as final optical densities >100 can be achieved. Therefore, in this study, we developed a suitable protocol for HCD cultivations of ApMTAP. Initially, optimum expression conditions were determined in 24-well plates using a fed-batch medium. Subsequently, HCD cultivations were performed using E. coli BL21-Gold cells, by employing a glucose-limited fed-batch strategy. Comparing different growth rates in stirred-tank bioreactors, cultivations revealed that growth at maximum growth rates until induction resulted in the highest yields of ApMTAP. On a 500-mL scale, final cell dry weights of 87.1–90.1 g L−1 were observed together with an overproduction of ApMTAP in a 1.9%–3.8% ratio of total protein. Compared to initially applied shake flask cultivations with terrific broth (TB) medium the volumetric yield increased by a factor of 136. After the purification of ApMTAP via heat treatment and affinity chromatography, a purity of more than 90% was determined. Activity testing revealed specific activities in the range of 0.21 ± 0.11 (low growth rate) to 3.99 ± 1.02 U mg−1 (growth at maximum growth rate). Hence, growth at maximum growth rate led to both an increased expression of the target protein and an increased specific enzyme activity. This study paves the way towards the application of thermostable nucleoside phosphorylases in industrial applications due to an improved heterologous expression in Escherichia coli. 相似文献
12.
《Bioscience, biotechnology, and biochemistry》2013,77(7):1248-1255
Using an inosine-producing mutant of Escherichia coli, the contributions of the central carbon metabolism for overproducing inosine were investigated. Sodium gluconate instead of glucose was tested as a carbon source to increase the supply of ribose-5-phosphate through the oxidative pentose phosphate pathway. The edd (6-phosphogluconate dehydrase gene)-disrupted mutant accumulated 2.5 g/l of inosine from 48 g/l of sodium gluconate, compared with 1.4 g/l of inosine in the edd wild strain. The rpe (ribulose phosphate 3-epimerase gene)-disrupted mutant resulted in low cell growth and low inosine production on glucose and on gluconate. The disruption of pgi (glucose-6-phosphate isomerase gene) was effective for increasing the accumulation of inosine from glucose but resulted in low cell growth. The pgi-disrupted mutant accumulated 3.7 g/l of inosine from 40 g/l of glucose when 8 g/l of yeast extract was added to the medium. Furthermore, to improve effective utilization of adenine, the yicP (adenine deaminase gene)-disrupted mutant was evaluated. It showed higher inosine accumulation, of 3.7 g/l, than that of 2.8 g/l in the yicP wild strain when 4 g/l of yeast extract was added to the medium. 相似文献
13.
E. V. Tret’yakova E. V. Salimova L. V. Parfenova 《Russian Journal of Bioorganic Chemistry》2018,44(5):547-552
An effective method of a modification of the anhydride ring of the maleopimaric acid methyl ester by means of the cyanoethylation reaction was developed. A primary screening of a cytotoxic activity in vitro demonstrated an ability of the cyanoethyl derivative of the maleopimaric acid methyl ester to induce the death of the PC-3 cells of prostate cancer. 相似文献
14.
New norcantharidin analogs were designed and obtained as compounds with biological activity. As a starting material, exo‐7‐oxabicyclo[2.2.1]heptane‐2,3‐dicarboxylic acid anhydride was used. Three groups of compounds: dicarboximides, triazoles and thiazolidines were obtained in multistep reactions. The 1H‐ and 13C‐NMR spectra were used to confirm the structures of all obtained products and they were in agreement with the proposed structure of substances. All derivatives were screened for their antioxidant activity. The most promising group was dicarboximides ( 1 – 4 , 6 ). Derivatives 2–4 displayed antioxidant activity with EC50=7.75–10.89 μg/ml, which may be comparable to strong antioxidant Trolox (EC50=6.13 μg/ml). Excellent activity with EC50=10.75 μg/ml also presented norcantharidin analog with 1,2,4‐triazole system ( 12 ). 相似文献
15.
目的:构建表达4-羟基苯乙酸-3-羟化酶A(4-hydroxyphenylacetate-3-hydroxylase A,HHA)的重组菌株,进而以酪醇为底物,利用重组菌株转化生产羟基酪醇。方法:选择大肠杆菌BL21(DE3)为模板来扩增HHA基因,经酶切后连接到表达载体p ET-28a中,获得重组表达载体p ET28a-HHA,将重组载体转化到感受态细胞BL21(DE3),在重组菌液中加入适量酪醇,利用胞内的重组酶对酪醇加羟基合成羟基酪醇,细胞离心后,分别采用薄层层析法和气质联用法检测上清液中羟基酪醇的转化结果。结果:IPTG诱导表达后,经SDS-PAGE分析获得分子质量分别为58.8k Da和18.5k Da的两条蛋白质条带。薄层层析法和气质联用法均检测到催化产物羟基酪醇的生成。结论:成功构建了表达HHA的重组菌株,该菌株可有效将酪醇转化为羟基酪醇。 相似文献
16.
Xuandi Zhang Guo‐Ning Zhang Yujia Wang Mei Zhu Juxian Wang Ziqiang Li Donghui Li Shan Cen Yucheng Wang 《化学与生物多样性》2019,16(2)
Influenza A virus (IAV), a highly pathogenic virus to human beings, is most susceptible to mutation and thus causes rapid, severe global pandemics resulting in millions of fatalities worldwide. Since resistance to the existing anti‐influenza drugs is developing, innovative inhibitors with a different mode of action are urgently needed. The lead compound 6092B‐E5 has proven to be an effective antiviral reagent in our previous work. Using the principles of substitution and bioisosterism of the indole ring, six series of novel anti‐IAV target products were designed, synthesized and evaluated for their antiviral effect in this work. Compounds D1 , D3 , D9 , G1 , G3 , G12 and G23 were identified as promising anti‐IAV candidates with excellent anti‐IAV efficacy (IC50 values of 3.06–5.77 μm ) and low cytotoxicity (CC50 values up to and beyond 100 μm ). This work represents a successful application of the substitution and bioisosteric replacement strategy for the discovery of novel antiviral molecules that can be used for further structural optimization. 相似文献
17.
以芳香氰基化合物为起始原料,经胺解、乙酰化、催化氢化、关环等步骤得到新型二氢嘧啶类化合物14个,结构通过核磁共振氢谱、质谱、元素分析确证,并利用HepG2.2.15细胞进行了体外抗HBV活性评价.结果表明部分化合物具有明显的抗HBVDNA复制作用,其中化合物5c和5n抑制HBVDNA复制的半数有效浓度分别为0.87μmol/L和0.67μmol/L.初步探讨了活性化合物的构效关系,同时运用分子排阻色谱法考察了活性化合物与病毒衣壳蛋白的相互作用. 相似文献
18.
Enhanced Biological Hydrogen Production from Escherichia coli with Surface Precipitated Cadmium Sulfide Nanoparticles
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Bo Wang Cuiping Zeng Ka Him Chu Dan Wu Ho Yin Yip Liqun Ye Po Keung Wong 《Liver Transplantation》2017,7(20)
The precipitation of cadmium sulfide nanoparticles is induced on the surface of Escherichia coli , and the biological hydrogen production efficiency under visible light (VL) irradiation is investigated. When endogenous [Ni–Fe]‐hydrogenase is anaerobically induced, an additional 400 µmol of hydrogen gas is generated within 3 h from the hybrid system suspension (50 mL) under VL irradiation (2000 W m?2), corresponding to an increase in hydrogen production of ≈30%. The apparent quantum efficiencies of the hybrid system under 470 and 620 nm VL irradiation are 7.93% and 9.59%, respectively, which are higher than those of many photoheterotrophic bacteria. Furthermore, the mechanism of the enhanced hydrogen evolution is investigated. The interaction between photogenerated electrons and cells of E. coli is confirmed by heat‐treatment, electron‐scavenger, and separation studies. The acceleration of pyruvate generation, inhibition of lactate fermentation, increase of formate concentration, stimulation of hydrogenase activity, and elevation of nicotinamide adenine dinucleotide (NAD)H/NAD ratio in the hybrid system are responsible for the enhanced hydrogen production. A feasibility study is also conducted using wastewater and natural sunlight for the hydrogen production by the hybrid system. An additional 120 µmol of hydrogen is generated from the hybrid system within 3 h under these conditions using natural resources. 相似文献
19.
鲑鱼降钙素及其类似物的合成 总被引:1,自引:0,他引:1
鲑鱼降钙素及其类似物的合成潘和平王良友陈正英王芳王会信(军事医学科学院基础医学研究所,北京100850)SynthesisofSalmonCalcitoninandItsAnalogsPanHe-PingWangLiang-YouChenZheng-... 相似文献
20.
Mikhaleva I. I. Prudchenko I. A. Onoprienko L. V. Lobanov A. V. Rodionov A. N. Tukhovskaya E. A. Khokhlova O. N. Shykhutdinova E. R. Murashev A. N. Ivanov V. T. 《Russian Journal of Bioorganic Chemistry》2018,44(6):631-639
Russian Journal of Bioorganic Chemistry - We performed a computer search for the DSIP homologous structures in the protein data bases and found the KND peptide (WKGGDNASGE), which was closely... 相似文献