Production of a Recombinant Laccase from Pichia pastoris and Biodegradation of Chlorpyrifos in a Laccase/Vanillin System

( Hui fang Xie ),( Qi Li ),( Min Min Wang ),( Lin Guo Zhao ) 한국미생물 · 생명공학회 2013 Journal of microbiology and biotechnology Vol.23 No.6

The recombinant strain P. pastoris GS115-lccC was used to produce laccase with high activity. Factors influencing laccase expression, such as pH, methanol concentration, copper concentration, peptone concentration, shaker rotate speed, and medium volume were investigated. Under the optimal conditions, laccase activity reached 12,344 U/L on day 15. The recombinant enzyme was purified by precipitating and dialyzing to electrophoretic homogeneity, and was estimated to have a molecular mass of about 58 kDa. When guaiacol was the substrate, the laccase showed the highest activity at pH 5.0 and was stable when the pH was 4.5~6.0. The optimal temperature for the laccase to oxidize guaiacol was 60℃, but it was not stable at high temperature. The enzyme could remain stable at 30℃ for 5 days. The recombinant laccase was used to degrade chlorpyrifos in several laccase/mediator systems. Among three synthetic mediators (ABTS, HBT, VA) and three natural mediators (vanillin, 2,6-DMP, and guaiacol), vanillin showed the most enhancement on degradation of chlorpyrifos. Both laccase and vanillin were responsible for the degradation of chlorpyrifos. A higher dosage of vanillin may promote a higher level of degradation of chlorpyrifos, and the 2-step addition of vanillin led to 98% chlorpyrifos degradation. The degradation of chlorpyrifos was faster in the L/V system (kobs = 0.151) than that in the buffer solution (kobs = 0.028).

Changes in Protein Phosphorylation during Salivary Gland Degeneration in Haemaphysalis longicornis

Qi Xiao,Yuhong Hu,Xiaohong Yang,Jianna Tang,Xiaoshuang Wang,Xiaomin Xue,Mengxue Li,Minjing Wang,Yinan Zhao,Jingze Liu,Hui Wang 대한기생충학ㆍ열대의학회 2020 The Korean Journal of Parasitology Vol.58 No.2

The ticks feed large amount of blood from their hosts and transmit pathogens to the victims. The salivary gland plays an important role in the blood feeding. When the female ticks are near engorgement, the salivary gland gradually loses its functions and begins to rapidly degenerate. In this study, data-independent acquisition quantitative proteomics was used to study changes in the phosphorylation modification of proteins during salivary gland degeneration in Haemaphysalis longicornis. In this quantitative study, 400 phosphorylated proteins and 850 phosphorylation modification sites were identified. Trough RNA interference experiments, we found that among the proteins with changes in phosphorylation, apoptosis-promoting Hippo protein played a role in salivary gland degeneration.

A review of experimental methods for characterising composite viscosities of continuous fibre-reinforced polymer composites

Wang Jinhuo,Han Yang,Ge Xiaohong,Qi Zhengbing,Zhao Jun,Wang Rongwen,Wu Huawei,Han Taiping,Sun Shaoxun,Wang Hui,Lin Jia,Liu Yuejun,Kong Xiangsong,Chen Qiming,Zeng Xiangxu 한국유변학회 2023 Korea-Australia rheology journal Vol.35 No.2

Optimisation design of composite structures requires an accurate predictive model for forming behaviour. The simulation process contains a number of model parameters which include transverse and longitudinal viscosities of continuous fibrereinforced viscous composites, fundamental to predicting the shear rheology. Shearing the unidirectional composite along the fibre direction gives a measure of the longitudinal viscosity (LV), whilst shearing across or transverse to the fibre direction gives a measure of the transverse viscosity (TV). Numerous experimental work was conducted in the past to measure these two viscosities for various materials. However, conflicting measurements by different test methods were obtained and these apparent discrepancies had not yet been systematically investigated in any single study. This paper reviews previous work on characterisation techniques to further understand the cause of such discrepancy, and hence to improve measurement accuracy, which would benefit future work on theoretical modelling of the composite viscosities and optimisation simulation of composites forming. Some important findings, such as effects of resin-rich areas, contributory factors of elastic effects, non-Newtonian behaviour for composites with Newtonian matrix, aspect ratio and end effects of test samples, geometry effects of fibres and fibre rearrangement during shearing, existence of a mathematical relationship between LV and TV and necessary benchmarking exercise using Newtonian matrix composites, were summarised.

Identification of Dominant Microbial Community and Diversity in Continuously Cropped Pepper Fields

Hui Wang,Li Liu,Shuyi Yu,Tianshu Guan,Baihong Li,Qi Wang,Changyuan Liu 한국원예학회 2021 원예과학기술지 Vol.39 No.4

Pepper blight is the most significant soil-borne disease affecting the continuous cropping of peppers. To identify the effect of Phytophthora capsici infection on microbial flora, we isolated and counted the microorganisms collected from the rhizosphere soil of P. capsici-affected farms that continuously cropped pepper for 3, 6, and 9 years in Liaoning Province, China. The colony and cell morphology, physiological and biochemical characteristics, and 16S rDNA sequence of bacteria and actinomycetes were documented. In addition, colony and microscopic morphology of fungi and the rDNA-ITS sequence were analysed for classification. We observed that healthy and diseased peppers had the largest number of bacteria in the rhizosphere followed by actinomycetes and fungi. After infection, the number of bacteria and actinomycetes decreased with a corresponding increase in the number of fungi, leading to a reduction in the ratio of bacteria/fungi to actinomycetes/fungi. We identified 15 dominant bacterial strains, of which Bacillus represented the most abundant genus consisting of 7 strains followed by Flavobacterium and Staphylococcus. Furthermore, 15 of the 17 actinomycetes strains belonged to the genus Streptomyces. Among the six fungal strains, we found P. infestans, Fusarium, and Penicillium consisting of two strains each. This study elucidated the impact of pathogenic P. capsici on the composition of soil microbes over time and characterized several cultivatable dominant bacterial groups, which can provide a basis for practical intervention strategies to improve soil conditions for continuous cropping.

Elevated PIVKA-II is Associated with Early Recurrence and Poor Prognosis in BCLC 0-A Hepatocellular Carcinomas

Wang, Bei-Li,Tan, Qi-Wen,Gao, Xing-Hui,Wu, Jiong,Guo, Wei Asian Pacific Journal of Cancer Prevention 2014 Asian Pacific journal of cancer prevention Vol.15 No.16

Background: To investigate the prognostic value of serum PIVKA-II (prothrombin induced by the absence of vitamin K or antagonist-II) in BCLC (Barcelona Clinic Liver Cancer) 0-A hepatocellular carcinoma (HCC) patients after curative resection. Materials and Methods: Preoperative sera were collected from 140 patients with BCLC 0-A HCCs undergoing curative resection during 2011-2012 in Zhongshan Hospital. Follow-up ended on November 2013. ELISA was used to detect the serum concentrations of preoperative PIVKA-II. The prognostic value of PIVKA-II and other clinicopathological factors was analyzed by the Kaplan-Meier method and the multivariate Cox proportional hazards model. Results: During follow-up, 39 of 140 patients suffered recurrence and the 1-year recurrence rate was 27.9%. The high-PIVKA-II expression group had lower 1-year time to progression (TTP) compared with the low-expression group (54.8% vs 20.2%, p<0.001). Patients with high preoperative PIVKA-II expression showed a relatively higher risk of developing postoperative recurrence than those with low expression in the low-recurrence-risk subgroups, including ${\alpha}$-fetoprotein ${\leq}400ng/mL$ (45.4% vs 16.7%; p=0.006), tumor size ${\leq}5cm$ (54.2% vs 18.1%; p<0.001), single tumor (56.0% vs 19.1%; p<0.001), absence of satellite lesions (53.3% vs 19.8%; p=0.001), absence of vascular invasion (52.6% vs 14.9%; p=0.002), and Edmondson stage I/II (60.9% vs 20.3%; p<0.001). PIVKA-II was the strongest independent prognostic factor for TTP (hazard ratio, 2.877; 95% CI 1.524-5.429; p=0.001). Conclusions: Elevated PIVKA-II is associated with early recurrence of BCLC 0-A HCC after curative resection and can be considered a novel prognostic predictor.

Detection for Tobacco Mosaic Virus and Potato Virus Y by DNA Chips Techniques

Hui Jia,Jin-Zhong Wang,Si-yuan Wen,Sheng-Qi Wang,Jin-Gao Dong,Min-Zhao Zhang 한국원예학회 2006 한국원예학회 기타간행물 Vol.- No.-

WDRP, a DWD protein component of CUL4-based E3 ligases, acts as a receptor of CDPK-related protein kinase 5 to mediate kinase degradation in Arabidopsis

Hui-Juan Teng,Yue Guo,Jian-Qi Wang,Rongjun Li,Ying-Tang Lu,Lei Zhang 한국식물학회 2016 Journal of Plant Biology Vol.59 No.6

CRK5 is a member of the Arabidopsis thaliana Ca2+-dependent protein kinase-related kinase family. Here, a yeast two-hybrid screen was performed with a truncated form of AtCRK5 as bait to identify interacting proteins and determine its physiological roles. One gene encoding the DWD protein WDRP was isolated. Furthermore, in vitro and in vivo co-immunoprecipitation results strongly supported that these two proteins interact with each other. Using a cell-free degradation assay, we also established that CRK5 was an unstable protein that was degraded through the proteasome pathway. The rate of CRK5 degradation was delayed in a WDRP knockout line. On the other hand, the degradation of CRK5 mediated by WDRP might not affect the phosphorylation of PIN2 by CRK5. Overall, we demonstrated that AtCRK5 interacted with a DWD protein, AtWDRP; the protein AtWDRP targets the kinase for ubiquitin-dependent degradation. Therefore, this report describes a new kinase regulation pathway for CRK family proteins in Arabidopsis.

Recombinant Human Thioredoxin-1 Protects Macrophages from Oxidized Low-Density Lipoprotein-Induced Foam Cell Formation and Cell Apoptosis

( Hui Zhang ),( Qi Liu ),( Jia-le Lin ),( Yu Wang ),( Ruo-xi Zhang ),( Jing-bo Hou ),( Bo Yu ) 한국응용약물학회 2018 Biomolecules & Therapeutics(구 응용약물학회지) Vol.26 No.2

Oxidized low-density lipoprotein (ox-LDL)-induced macrophage foam cell formation and apoptosis play critical roles in the pathogenesis of atherosclerosis. Thioredoxin-1 (Trx) is an antioxidant that potently protects various cells from oxidative stress-induced cell death. However, the protective effect of Trx on ox-LDL-induced macrophage foam cell formation and apoptosis has not been studied. This study aims to investigate the effect of recombinant human Trx (rhTrx) on ox-LDL-stimulated RAW264.7 macrophages and elucidate the possible mechanisms. RhTrx significantly inhibited ox-LDL-induced cholesterol accumulation and apoptosis in RAW264.7 macrophages. RhTrx also suppressed the ox-LDL-induced overproduction of lectin-like oxidized LDL receptor (LOX- 1), Bax and activated caspase-3, but it increased the expression of Bcl-2. In addition, rhTrx markedly inhibited the ox-LDL-induced production of intracellular reactive oxygen species (ROS) and phosphorylation of p38 mitogen-activated protein kinases (MAPK). Furthermore, anisomycin (a p38 MAPK activator) abolished the protective effect of rhTrx on ox-LDL-stimulated RAW264.7 cells, and SB203580 (a p38 MAPK inhibitor) exerted a similar effect as rhTrx. Collectively, these findings indicate that rhTrx suppresses ox-LDL-stimulated foam cell formation and macrophage apoptosis by inhibiting ROS generation, p38 MAPK activation and LOX-1 expression. Therefore, we propose that rhTrx has therapeutic potential in the prevention and treatment of atherosclerosis.

Whole-genome resequencing reveals domestication and signatures of selection in Ujimqin, Sunit, and Wu Ranke Mongolian sheep breeds

Wang Hanning,Zhong Liang,Dong Yanbing,Meng Lingbo,Ji Cheng,Luo Hui,Fu Mengrong,Qi Zhi,Mi Lan 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.9

Objective: The current study aimed to perform whole-genome resequencing of Chinese indigenous Mongolian sheep breeds including Ujimqin, Sunit, and Wu Ranke sheep breeds (UJMQ, SNT, WRK) and deeply analyze genetic variation, population structure, domestication, and selection for domestication traits among these Mongolian sheep breeds.Methods: Blood samples were collected from a total of 60 individuals comprising 20 WRK, 20 UJMQ, and 20 SNT. For genome sequencing, about 1.5 μg of genomic DNA was used for library construction with an insert size of about 350 bp. Pair-end sequencing were performed on Illumina NovaSeq platform, with the read length of 150 bp at each end. We then investigated the domestication and signatures of selection in these sheep breeds.Results: According to the population and demographic analyses, WRK and SNT populations were very similar, which were different from UJMQ populations. Genome wide association study identified 468 and 779 significant loci from SNT vs UJMQ, and UJMQ vs WRK, respectively. However, only 3 loci were identified from SNT vs WRK. Genomic comparison and selective sweep analysis among these sheep breeds suggested that genes associated with regulation of secretion, metabolic pathways including estrogen metabolism and amino acid metabolism, and neuron development have undergone strong selection during domestication.Conclusion: Our findings will facilitate the understanding of Chinese indigenous Mongolian sheep breeds domestication and selection for complex traits and provide a valuable genomic resource for future studies of sheep and other domestic animal breeding. Objective: The current study aimed to perform whole-genome resequencing of Chinese indigenous Mongolian sheep breeds including Ujimqin, Sunit, and Wu Ranke sheep breeds (UJMQ, SNT, WRK) and deeply analyze genetic variation, population structure, domestication, and selection for domestication traits among these Mongolian sheep breeds. Methods: Blood samples were collected from a total of 60 individuals comprising 20 WRK, 20 UJMQ, and 20 SNT. For genome sequencing, about 1.5 μg of genomic DNA was used for library construction with an insert size of about 350 bp. Pair-end sequencing were performed on Illumina NovaSeq platform, with the read length of 150 bp at each end. We then investigated the domestication and signatures of selection in these sheep breeds. Results: According to the population and demographic analyses, WRK and SNT populations were very similar, which were different from UJMQ populations. Genome wide association study identified 468 and 779 significant loci from SNT vs UJMQ, and UJMQ vs WRK, respectively. However, only 3 loci were identified from SNT vs WRK. Genomic comparison and selective sweep analysis among these sheep breeds suggested that genes associated with regulation of secretion, metabolic pathways including estrogen metabolism and amino acid metabolism, and neuron development have undergone strong selection during domestication. Conclusion: Our findings will facilitate the understanding of Chinese indigenous Mongolian sheep breeds domestication and selection for complex traits and provide a valuable genomic resource for future studies of sheep and other domestic animal breeding.

Learning-Based Multiple Pooling Fusion in Multi-View Convolutional Neural Network for 3D Model Classification and Retrieval

Hui Zeng,Qi Wang,Chen Li,Wei Song 한국정보처리학회 2019 Journal of information processing systems Vol.15 No.5

We design an ingenious view-pooling method named learning-based multiple pooling fusion (LMPF), andapply it to multi-view convolutional neural network (MVCNN) for 3D model classification or retrieval. By thismeans, multi-view feature maps projected from a 3D model can be compiled as a simple and effective featuredescriptor. The LMPF method fuses the max pooling method and the mean pooling method by learning a setof optimal weights. Compared with the hand-crafted approaches such as max pooling and mean pooling, theLMPF method can decrease the information loss effectively because of its “learning” ability. Experiments onModelNet40 dataset and McGill dataset are presented and the results verify that LMPF can outperform thoseprevious methods to a great extent.