Biochemical characterization and substrate profiling of a new NADH-dependent enoate reductase from Lactobacillus casei

Carbon-carbon double bond of α,β-unsaturated carbonyl compounds can be reduced by enoate reductase (ER), which is an important reaction in fine chemical synthesis. A putative enoate reductase gene from Lactobacillus casei str. Zhang was cloned into pET-21a+ and expressed in Escherichia coli BL21 (DE3) host cells. The encoded enzyme (LacER) was purified by ammonium sulfate precipitation and treatment in an acidic buffer. This enzyme was identified as a NADH-dependent enoate reductase, which had a K(m) of 0.034 ± 0.006 mM and k(cat) of (3.2 ± 0.2) × 103 s?1 toward NADH using 2-cyclohexen-1-one as the substrate. Its K(m) and k(cat) toward substrate 2-cyclohexen-1-one were 1.94 ± 0.04 mM and (8.4 ± 0.2) × 103 s?1, respectively. The enzyme showed a maximum activity at pH 8.0-9.0. The optimum temperature of the enzyme was 50-55°C, and LacER was relatively stable below 60 °C. The enzyme was active toward aliphatic alkenyl aldehyde, ketones and some cyclic anhydrides. Substituted groups of cyclic α,β-unsaturated ketones and its ring size have positive or negative effects on activity. (R)-(-)-Carvone was reduced to (2R,5R)-dihydrocarvone with 99% conversion and 98% (diasteromeric excess: de) stereoselectivity, indicating a high synthetic potential of LacER in asymmetric synthesis.     

中国长头网蝽属记述（半翅目:网蝽科）

记述中国分布的长头网蝽属昆虫5种,其中包括1新种,鄂长头网蝽Cantacader hubeianus sp.nov., 2个中国新纪录种,日本长头网蝽 C. japanicus Drake, 1947和五脊长头网蝽 C. quinquecostatus (Fieber, 1844)。给出了长头网蝽属中国种类检索表和23幅形态图。模式标本保存于南开大学昆虫标本室。     

The deubiquitinating enzyme USP1 regulates the Fanconi anemia pathway

Protein ubiquitination and deubiquitination are dynamic processes implicated in the regulation of numerous cellular pathways. Monoubiquitination of the Fanconi anemia (FA) protein FANCD2 appears to be critical in the repair of DNA damage because many of the proteins that are mutated in FA are required for FANCD2 ubiquitination. By screening a gene family RNAi library, we identify the deubiquitinating enzyme USP1 as a novel component of the Fanconi anemia pathway. Inhibition of USP1 leads to hyperaccumulation of monoubiquitinated FANCD2. Furthermore, USP1 physically associates with FANCD2, and the proteins colocalize in chromatin after DNA damage. Finally, analysis of crosslinker-induced chromosomal aberrations in USP1 knockdown cells suggests a role in DNA repair. We propose that USP1 deubiquitinates FANCD2 when cells exit S phase or recommence cycling after a DNA damage insult and may play a critical role in the FA pathway by recycling FANCD2.     

Elevated O3 reduces the fitness of Bemisia tabaci via enhancement of the SA-dependent defense of the tomato plant

The effect of elevated O₃ on tomato plants of three different genotypes (wild-type, a jasmonic acid (JA) defense-enhanced genotype (35S) and a JA-deficient genotype (spr2)) grown in association with the whitefly Bemisia tabaci Gennadius biotype B was examined in the field in open-top chambers. We experimentally tested the hypothesis that elevated O₃ tends to reduce the nutrition of tomato plants, and to increase the SA-dependent pathway defenses and the secondary metabolites, and therefore decrease the population fitness of the whitefly. The results show that for all three tomato genotypes, elevated O₃ reduced the soluble sugars and free amino acids, increased the phenylalanine ammonia-lyase enzyme activity and the accumulated salicylic acid (SA), and up-regulated the pathogenesis-related protein (PR1), which is commonly considered to be the whitefly-resistance gene product involved in SA-dependent defense. Elevated O₃ did not affect the JA level in any of the three plant genotypes, but it increased the levels of some secondary metabolites, including total phenolics and condensed tannins. Elevated O₃ prolonged the developmental time of whiteflies fed on the three plant genotypes, and it also reduced the fecundity and the intrinsic rate of increase of whiteflies fed on either the 35S or the wild-type plants. These results suggest that elevated O₃ reduces the nutrition of tomato plants and enhances their SA content, relative PR mRNA expression and secondary metabolism, resulting in decreased fitness of whiteflies on these tomato plants.     

Replication Study in a Japanese Population of Six Susceptibility Loci for Type 2 Diabetes Originally Identified by a Transethnic Meta-Analysis of Genome-Wide Association Studies

AimWe performed a replication study in a Japanese population to evaluate the association between type 2 diabetes and six susceptibility loci (TMEM154, SSR1, FAF1, POU5F1, ARL15, and MPHOSPH9) originally identified by a transethnic meta-analysis of genome-wide association studies (GWAS) in 2014.MethodsWe genotyped 7,620 Japanese participants (5,817 type 2 diabetes patients and 1,803 controls) for each of the single nucleotide polymorphisms (SNPs) using a multiplex polymerase chain reaction invader assay. The association of each SNP locus with the disease was evaluated using logistic regression analysis.ResultsOf the six SNPs examined in this study, four (rs6813195 near TMEM154, rs17106184 in FAF1, rs3130501 in POU5F1 and rs4275659 near MPHOSPH9) had the same direction of effect as in the original reports, but two (rs9505118 in SSR1 and rs702634 in ARL15) had the opposite direction of effect. Among these loci, rs3130501 and rs4275659 were nominally associated with type 2 diabetes (rs3130501; p = 0.017, odds ratio [OR] = 1.113, 95% confidence interval [CI] 1.019–1.215, rs4275659; p = 0.012, OR = 1.127, 95% CI 1.026–1.238, adjusted for sex, age and body mass index), but we did not observe a significant association with type 2 diabetes for any of the six evaluated SNP loci in our Japanese population.ConclusionsOur results indicate that effects of the six SNP loci identified in the transethnic GWAS meta-analysis are not major among the Japanese, although SNPs in POU5F1 and MPHOSPH9 loci may have some effect on susceptibility to type 2 diabetes in this population.     

人心肌肌球蛋白轻链1的克隆,表达纯化和单抗制备

报道了中国人心肌肌球蛋白轻链１ｃＤＮＡ的核苷酸序列,并由此推算的氨基酸序列。与国外发表的人心肌肌球蛋白轻链的氨基酸序列比较,发现有两处差异,即在２４位,由谷氨酸变为丙氨酸,则从９８位起至１０１位有４个氨基酸序列的连续差异,即由天冬酰胺－精氨酸－丝氨酸－赖氨酸变为赖氨酸－脯氨酸－精氨酸－谷氨酰妥,推测可能是由于人种差异而引起的。利用该ｃＤＮＡ在大肠杆菌内的表达产物,已获得一株高效的抗中国人心肌肌球蛋     

A Scheme to Optimize Flow Routing and Polling Switch Selection of Software Defined Networks

This paper aims at minimizing the communication cost for collecting flow information in Software Defined Networks (SDN). Since flow-based information collecting method requires too much communication cost, and switch-based method proposed recently cannot benefit from controlling flow routing, jointly optimize flow routing and polling switch selection is proposed to reduce the communication cost. To this end, joint optimization problem is formulated as an Integer Linear Programming (ILP) model firstly. Since the ILP model is intractable in large size network, we also design an optimal algorithm for the multi-rooted tree topology and an efficient heuristic algorithm for general topology. According to extensive simulations, it is found that our method can save up to 55.76% communication cost compared with the state-of-the-art switch-based scheme.