Review : Purification and Characterization of a Novel Extracellular Thermostable Alkaline Protease from Streptomyces sp. M30

( Yan Xin ),( Zhibin Sun ),( Qiongzhen Chen ),( Jue Wang ),( Yicheng Wang ),( Linfeng Luogong ),( Shuhuan Li ),( Weiliang Dong ),( Zhongli Cui ),( Yan Huang ) 한국미생물 · 생명공학회 2015 Journal of microbiology and biotechnology Vol.25 No.11

A novel alkaline protease from Streptomyces sp. M30, SapHM, was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and DEAE-Sepharose chromatography, with a yield of 15.5% and a specific activity of 29,070 U/mg. Tryptic fragments of the purified SapHM were obtained by electrospray ionization quadrupole timeof- flight mass spectrometry. Nucleotide sequence analysis revealed that the gene sapHM contained 1,179 bp, corresponding to 392 amino acids with conserved Asp156, His187, and Ser339 residues of alkaline protease. The first 24 amino acid residues were predicted to be a signal peptide, and the molecular mass of the mature peptide was 37.1 kDa based on amino acid sequences and mass spectrometry. Pure SapHM was optimally active at 80°C in 50 mM glycine-NaOH buffer (pH 9.0), and was broadly stable at 0-50°C and pH 4.0-9.0. The protease relative activity was increased in the presence of Ni2+, Mn2+, and Cu2+ to 112%, 113%, and 147% of control, respectively. Pure SapHM was also activated by dimethylformamide, dimethyl sulfoxide, Tween 80, and urea. The activity of the purified enzyme was completely inhibited by phenylmethylsulfonyl fluoride, indicating that it is a serine-type protease. The Km and Vmax values were estimated to be 35.7 mg/ml, and 5 × 104 U/mg for casein. Substrate specificity analysis showed that SapH was active on casein, bovine serum albumin, and bovine serum fibrin.

A Novel Bi-directional Promoter Cloned from Melon and Its Activity in Cucumber and Tobacco

( Cui Yan Wang ),( Dong Feng Ding ),( Rui Xiang Yan ),( Xiao Ju Yu ),( Wei Dong Li ),( Ming Gang Li ) 한국식물학회 2008 Journal of Plant Biology Vol.51 No.2

A bi-directional promoter, DP, was cloned by PCR amplification using the genomic DNA of melon as template. Analysis of its cis-acting elements in both directions revealed a series of inducible regulatory elements and some enhancer elements. To evaluate its transcriptional activity, DP in both directions was then cloned into vector pBI121 to replace the CaMV 35S promoter. DP in both directions also was inserted downstream of CaMV 35S to investigate whether the double promoter might affect expression of the uidA reporter gene at higher levels. Transient expression in cucumber leaves, stems, and fruits as well as in tobacco leaves and stems showed that DP in both directions drove transcription to much higher levels than did the single promoter CaMV 35S. However, activity of the double promoter was lower than the corresponding activity of the single promoter DP in both directions. These results demonstrate that DP is a natural bi-directional promoter, with much more activity than is found with the CaMV 35S promoter. Furthermore, in cucumber and tobacco, it is not suitable to insert DP in either direction downstream of the CaMV 35S promoter to form a double promoter.

Adaptive Virtual Impedance of Grid-Tied Inverters to Enhance the Stability in a Weak Grid

Yan Du,Qingqing Sun,Xiangzhen Yang,Linbo Cui,Jian Zhang,Fei Wang 대한전기학회 2019 Journal of Electrical Engineering & Technology Vol.14 No.3

In a weak grid, especially with a grid-voltage proportional feedforward controller, the grid-tied inverter tends to interact with the grid impedance and thus suff ers from poor power quality and even instability. In order to improve the stability, serial-parallel virtual impedance based on the derivative controllers of grid current and voltage is proposed in this paper. With the aim of optimizing the performance of the inverter for a variety of grid impedance, these derivative coeffi cients are adaptively tuned by the constraint of a minimum leading phase which is determined by the phase margin and the current tracking response. With this proposed adaptive virtual impedance, the grid-tied inverter can operate stably under a wide variety of grid impedance and voltage harmonic rejection performance is also improved. Simulation and experimental results validate the eff ectiveness of the proposed method and design methodology.

The Propagation and Construction of China’s National Image in 21st Century

Wang Weimin,Cui Yan 한국디지털정책학회 2011 디지털융복합연구 Vol.9 No.3

A Two-stage Stochastic Programming Model for Optimal Reactive Power Dispatch with High Penetration Level of Wind Generation

Cui, Wei,Yan, Wei,Lee, Wei-Jen,Zhao, Xia,Ren, Zhouyang,Wang, Cong The Korean Institute of Electrical Engineers 2017 Journal of Electrical Engineering & Technology Vol.12 No.1

The increasing of wind power penetration level presents challenges in classical optimal reactive power dispatch (ORPD) which is usually formulated as a deterministic optimization problem. This paper proposes a two-stage stochastic programming model for ORPD by considering the uncertainties of wind speed and load in a specified time interval. To avoid the excessive operation, the schedule of compensators will be determined in the first-stage while accounting for the costs of adjusting the compensators (CACs). Under uncertainty effects, on-load tap changer (OLTC) and generator in the second-stage will compensate the mismatch caused by the first-stage decision. The objective of the proposed model is to minimize the sum of CACs and the expected energy loss. The stochastic behavior is formulated by three-point estimate method (TPEM) to convert the stochastic programming into equivalent deterministic problem. A hybrid Genetic Algorithm-Interior Point Method is utilized to solve this large-scale mixed-integer nonlinear stochastic problem. Two case studies on IEEE 14-bus and IEEE 118-bus system are provided to illustrate the effectiveness of the proposed method.

Integration Sites and Genotype Distributions of Human Papillomavirus in Cervical Intraepithelial Neoplasia

Wang, Li,Dai, Shu-Zhen,Chu, Hui-Jun,Cui, Hong-Fei,Xu, Xiao-Yan Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.6

Objectives: To analyse HPV integration prevalence and genotype distributions in cervical intraepithelial neoplasia (CIN) in east part of China, furthermore to assess preferential sites for common HPV integrations and provide baseline information for cervical abnormality screening and prevention. Methods: Integration of HPV in 113 paraffin-embedded cervical intraepithelial neoplasia samples was assessed using Gencap technology in Key Laboratory of Biotechnologies in BGI-Shenzhen. Results: 64 samples were HPV-integrated and as the cervical lesions increased, the integration rate became higher significantly (P=0.002). Fifteen different HPV genotypes were detected, 14 high-risk (16, 18, 31, 33, 51, 52, 56, 58, 66, 68) and 1 low-risk (11). The most common genotypes were HPV-16, 58, 33, 52, 66, and 56. Thirteen patients had co-integration involving mainly HPV-16 and 58. The frequency of HPV gene disruption was higher in L1 and E1 genes than in other regions of the viral genomes. Conclusion: Some 56.6% of CIN lesions in Qingdao had HPV integrations, and 67.2% of HPV-integrated patients were HPV-16 and 58, more prone to be integrated in younger patients below 45 years old. There exist preferential sites for HPV-16 and HPV-58 integration, and they are more likely to be disrupted in the L1 and E1 loci.

Molecular and phylogenetic analysis of Anaplasma spp. in sheep and goats from six provinces of China

Yan Zhang,Yali Lv,Feifei Zhang,Wenjing Zhang,Jinhong Wang,Yanyan Cui,Rongjun Wang,Fuchun Jian,Longxian Zhang,Changshen Ning 대한수의학회 2016 Journal of Veterinary Science Vol.17 No.4

Members of the genus Anaplasma are important emerging tick-borne pathogens in both humans and animals in tropical and subtropical areas. Here, we investigated the presence of Anaplasma spp. in 621 sheep and 710 goats from six provinces of China. Polymerase chain reaction(PCR) and DNA sequencing were conducted to determine the prevalence of Anaplasma (A.) phagocytophilum, A. ovis and A. bovis targetingthe 16S ribosomal RNA or the major surface protein 4 gene. PCR revealed Anaplasma in 39.0% (240/621) of sheep and 45.5% (323/710)of goats. The most frequently detected species was A. ovis (88/621, 14.2% for sheep; 129/710, 18.2% for goats), followed by A. bovis (60/621,9.7% for sheep; 74/710, 10.4% for goats) and A. phagocytophilum (33/621, 5.3% for sheep; 15/710, 2.1% for goats). Additionally, eight sheepand 20 goats were found to be infected with three pathogens simultaneously. DNA sequencing confirmed the presence of these three Anaplasmaspecies in the investigated areas, and phylogenetic analysis indicated that there was geographic segregation to a certain extent, as well as arelationship between the host and cluster of A. ovis. The results of the present study provide valuable data that helps understand theepidemiology of anaplasmosis in ruminants from China.

A Novel Approach of Feature Extraction for Analog Circuit Fault Diagnosis Based on WPD-LLE-CSA

Wang, Yuehai,Ma, Yuying,Cui, Shiming,Yan, Yongzheng The Korean Institute of Electrical Engineers 2018 Journal of Electrical Engineering & Technology Vol.13 No.6

The rapid development of large-scale integrated circuits has brought great challenges to the circuit testing and diagnosis, and due to the lack of exact fault models, inaccurate analog components tolerance, and some nonlinear factors, the analog circuit fault diagnosis is still regarded as an extremely difficult problem. To cope with the problem that it's difficult to extract fault features effectively from masses of original data of the nonlinear continuous analog circuit output signal, a novel approach of feature extraction and dimension reduction for analog circuit fault diagnosis based on wavelet packet decomposition, local linear embedding algorithm, and clone selection algorithm (WPD-LLE-CSA) is proposed. The proposed method can identify faulty components in complicated analog circuits with a high accuracy above 99%. Compared with the existing feature extraction methods, the proposed method can significantly reduce the quantity of features with less time spent under the premise of maintaining a high level of diagnosing rate, and also the ratio of dimensionality reduction was discussed. Several groups of experiments are conducted to demonstrate the efficiency of the proposed method.

ODV-Associated Proteins of the <i>Pieris rapae</i> Granulovirus

Wang, Xiao-Feng,Zhang, Bao-Qin,Xu, Hai-Jun,Cui, Ying-Jun,Xu, Yi-Peng,Zhang, Min-Juan,Han, Yeon Soo,Lee, Yong Seok,Bao, Yan-Yuan,Zhang, Chuan-Xi American Chemical Society 2011 JOURNAL OF PROTEOME RESEARCH Vol.10 No.6

<P><I>Alphabaculovirus</I> (lepidopteran-specific nucleopolyhedroviruses, NPV) and <I>Betabaculovirus</I> (granuloviruses, GV) are two main genera of the family Baculoviridae. The virion proteomes of <I>Alphabaculovirus</I> have been well studied; however, the <I>Betabaculovirus</I> virion compositions remain unclear. <I>Pieris rapae</I> granulovirus (PrGV) can kill larvae of <I>P. rapae</I>, a worldwide and important pest of mustard family crops. In this study, the occlusion-derived virus (ODV)-associated proteins of PrGV were identified using three mass spectrometry (MS) approaches. The MS analyses demonstrated that 47 proteins were present in PrGV-ODV. Of the 47 PrGV-ODV proteins, 33 have homologues identified previously in other baculovirus ODV/BVs, whereas 14 (P10, Pr21, Pr29, Pr35, Pr42, Pr54, P45/48, Pr83, Pr84, Pr89, Pr92, Pr111, Pr114 and FGF3) were newly identified ODV proteins. Seven of the 14 newly identified ODV proteins are specific to <I>Betabaculovirus</I>, including Pr35, Pr42, Pr54, Pr83, Pr84, Pr111 and Pr114. Furthermore, the data derived from these MS approaches were validated by immunoblotting analysis using antisera prepared from 11 randomly selected recombinant PrGV-ODV proteins (including 5 <I>Betabaculovirus</I>-unique proteins). Comparison analyses revealed the similar and different compositions between <I>Betabaculovirus</I> and <I>Alphabaculovirus</I> virions, which deepen our understanding of the baculovirus virion structure and provide helpful information on <I>Betabaculovirus</I>–host interaction studies.</P><P>We used three mass spectrometry (MS) approaches to identify the occlusion-derived virus (ODV)-associated proteins of the <I>Pieris rapae</I> Granulovirus. A total of 47 proteins were identified; 14 of them were first identified in the ODV, and 7 are specific to Granulovirus.</P><P><B>Graphic Abstract</B> <IMG SRC='http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/jprobs/2011/jprobs.2011.10.issue-6/pr2000804/production/images/medium/pr-2011-000804_0002.gif'></P>

Diagnostic Value of Superoxide Dismutase in Tuberculous and Malignant Pleural Effusions

Wang, Xin-Feng,Wu, Yan-Hua,Jiao, Jin,Guan, Cui-Ping,Yang, Xiao-Guang,Wang, Mao-Shui Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.2

The aim of this study was to investigate the diagnostic value of superoxide dismutase (SOD) in tuberculous pleural effusions (TPEs) and malignant pleural effusions (MPEs). Pleural effusion (PE) samples from 100 patients were classified on the basis of diagnosis as TPE (n=57) and MPE (n=43). The activity of SOD was determined by pyrolgallol assay. A significant difference was observed in SOD activity (P<0.01) between TPE and MPE, levels of being significantly higher in TPE compared to MPE. With a threshold value of 41 U/L, the area under the ROC curve was 0.653, SOD had a sensitivity of 61.4% and a specificity of 61.0% for differential diagnosis. Thus, SOD activity in PE was not a good biomarker in differentiating TPE and MPE. To the best of our knowledge, five SOD isoforms may be present in PE. Identification of which SOD contributes to the difference of SOD level between TPE and MPE is very important for illustrating mechanisms and improving the differential diagnostic value.