Engineering a Highly Thermostable and Stress Tolerant Superoxide Dismutase by N-terminal Modification and Metal Incorporation

Mingchang Li,Shuyi Guo,Xiaomin Li,Quan Wang,Lin Zhu,Chunyan Yin,Wei Wang 한국생물공학회 2017 Biotechnology and Bioprocess Engineering Vol.22 No.6

Thermophilic or hyperthermophilic SODs (superoxide dismutase) usually offer substantial biotechnological advantages over mesophilic SODs. Previously a 244-amino acid N-terminal domain (NTD) from a heatresistant SOD of Geobacillus thermodenitrificans NG80-2 was discovered and demonstrated to be able to confer thermostability to homologous mesophilic SODs, which revealed a new type of heat resistance mechanism. To further improve the heat resistance and stress tolerance of thermophilic cambialistic superoxide dismutase (Fe/Mn- SODAp) from Aeropyrum pernix K1 through metal incorporation and fusion with the newly found peptide NTD for broadening its industrial application, the wildtype SODAp and NTD-fused ntdSODAp were expressed in E. coli BL21 and incorporated with metal cofactors by two ways. Recombinant fusion SOD obtained by in vitro reconstitution (Mn-rec ntdSODAp) exhibited improved optimum temperature at 70oC and dramatically enhanced thermostability especially at 110oC with enhanced pH stability from 4 to 10 and higher tolerance for denaturants and organic media than Mn-rec SODAp. To the best of our knowledge, Mn-rec ntdSODAp could be the most heat resistant SOD. In addition, metal incorporation of SODAp and ntdSODAp via in vivo modification have been developed and proved to be more practical for industrial use. These results indicate that fusion with NTD along with metal incorporation can generate superimposed effect and be applied to enhance the stability of cambialistic thermophilic SODs, thus providing a universal and convenient bioengineering method for generating extremely stable SODs.

The Attenuation Mechanism and Live Vaccine Potential of a Low-Virulence Edwardsiella ictaluri Strain Obtained by Rifampicin Passaging Culture

Wang Shuyi,Hao Jingwen,Yang Jicheng,Zhang Qianqian,Li Aihua 한국미생물·생명공학회 2023 Journal of microbiology and biotechnology Vol.33 No.2

The rifampicin-resistant strain E9-302 of Edwardsiella ictaluri strain 669 (WT) was generated by continuous passage on BHI agar plates containing increasing concentrations of rifampicin. E9-302 was attenuated significantly by 119 times to zebrafish Danio rerio compared to WT in terms of the 50% lethal dose (LD50). Zebrafish vaccinated with E9-302 via intraperitoneal (IP) injection at a dose of 1 × 103 CFU/fish had relative percentage survival (RPS) rates of 85.7% when challenged with wild-type E. ictaluri via IP 14 days post-vaccination (dpv). After 14 days of primary vaccination with E9-302 via immersion (IM) at a dose of 4 × 107 CFU/ml, a booster IM vaccination with E9-302 at a dose of 2 × 107 CFU/ml exhibited 65.2% RPS against challenge with wild-type E. ictaluri via IP 7 days later. These results indicated that the rifampicin-resistant attenuated strain E9-302 had potential as a live vaccine against E. ictaluri infection. A previously unreported amino acid site change at position 142 of the RNA polymerase (RNAP) β subunit encoded by the gene rpoB associated with rifampicin resistance was identified. Analysis of the whole-genome sequencing results revealed multiple missense mutations in the virulence-related genes esrB and sspH2 in E9-302 compared with WT, and a 189 bp mismatch in one gene, whose coding product was highly homologous to glycosyltransferase family 39 protein. This study preliminarily explored the molecular mechanism underlying the virulence attenuation of rifampicin-resistant strain E9-302 and provided a new target for the subsequent study of the pathogenic mechanism of E. ictaluri.

In vitro Antioxidant Activities of Proanthocyanidins Extracted from the Lotus Seedpod and Ameliorative Effects on Learning and Memory Impairment in Scopolamine-induced Amnesia Mice

Juan Xiao,Shuyi Li,Yong Sui,Xiaopeng Li,Qian Wu,Ruifen Zhang,Mingwei Zhang,Bijun Xie,Zhida Sun 한국식품과학회 2015 Food Science and Biotechnology Vol.24 No.4

The antioxidant activity of proanthocyanidins extracted from the lotus seedpods (LSPC) in vitro and ameliorative effects on memory impairment induced using scopolamine in mice were studied. 2,2-diphenyl-1- picrylhydrazyl (DPPH) radical scavenging ability, oxygen radical absorbance capacity, and cellular antioxidant activity assays revealed a potent LSPC antioxidant activity. Y-maze and step-down avoidance testing showed that LSPC administration at 30, 60, and 90 mg/kg ameliorated memory impairment. LSPC improved glutathione peroxidase and superoxide dismutase activities, inhibited activities of monoamine oxidase-B, total nitric oxide synthase, neuronal nitric oxide synthase, and acetylcholinesterase, and had no influence on inducible nitric oxide synthase and nitric oxide levels in the brain. LSPC ameliorated scopolamineinduced memory impairment based on improvement of the antioxidant system and cholinergic activity, which may be associated with potent antioxidant ability.

Crashworthiness design of bionic-shell thin-walled tube under axial impact

Lingyun Qin,Shuyi Yang,Hongzhou Li,Juchuan Dai,Guosheng Wang,Qihui Ling,Zhewu Chen 대한기계학회 2023 JOURNAL OF MECHANICAL SCIENCE AND TECHNOLOGY Vol.37 No.7

Bionic structures have been widely utilized in the crashworthiness design of thin-walled structures due to their superior energy absorption capabilities. This study constructed a bionic-shell thin-walled tube (BST) with excellent crashworthiness based on the structural bionic principle using the shell shape cross-section as the prototype. First, the theoretical model of the mean crushing force (MCF) for BST under axial compression was developed. An experiment was conducted and the reliability of the finite element model was verified. Then, the effects of structural parameters, such as the number of ribs, wall thickness, and inner tube diameter on the crashworthiness of the BST were investigated using the finite element method. Finally, to obtain the ideal configuration of structural parameters, the BST was optimized using the response surface method (RSM) with specific energy absorption (SEA) and crushing force efficiency (CFE) as the optimization objectives and peak crushing force (PCF) as the constraint condition. The results showed that the BST with six ribs exhibited the best crashworthiness under the same mass. The optimized BST-6 was found to have better energy absorption performance than the double circular tube (DCT) and the bionic-horsetail thin-walled tube (BHT). Compared with the DCT, the SEA and CFE increased by 35.15 % and 32.23 %, respectively.

A possible mechanism to the antidepressant-like effects of 20 (S)-protopanaxadiol based on its target protein 14-3-3 ζ

Chen, Lin,Li, Ruimei,Chen, Feiyan,Zhang, Hantao,Zhu, Zhu,Xu, Shuyi,Cheng, Yao,Zhao, Yunan The Korean Society of Ginseng 2022 Journal of Ginseng Research Vol.46 No.5

Background: Ginsenosides and their metabolites have antidepressant-like effects, but the underlying mechanisms remain unclear. We previously identified 14-3-3 ζ as one of the target proteins of 20 (S)-protopanaxadiol (PPD), a fully deglycosylated ginsenoside metabolite. Methods: Corticosterone (CORT) was administered repeatedly to induce the depression model, and PPD was given concurrently. The tail suspension test (TST) and the forced swimming test (FST) were used for behavioral evaluation. All mice were sacrificed. Golgi-cox staining, GSK 3β activity assay, and Western blot analysis were performed. In vitro, the kinetic binding analysis with the Biolayer Interferometry (BLI) was used to determine the molecular interactions. Results: TST and FST both revealed that PPD reversed CORT-induced behavioral deficits. PPD also ameliorated the CORT-induced expression alterations of hippocampal Ser9 phosphorylated glycogen synthase kinase 3β (p-Ser9 GSK 3β), Ser133 phosphorylated cAMP response element-binding protein (p-Ser133 CREB), and brain-derived neurotrophic factor (BDNF). Moreover, PPD attenuated the CORT-induced increase in GSK 3β activity and decrease in dendritic spine density in the hippocampus. In vitro, 14-3-3 ζ protein specifically bound to p-Ser9 GSK 3β polypeptide. PPD promoted the binding and subsequently decreased GSK 3β activity. Conclusion: These findings demonstrated the antidepressant-like effects of PPD on the CORT-induced mouse depression model and indicated a possible target-based mechanism. The combination of PPD with the 14-3-3 ζ protein may promote the binding of 14-3-3 ζ to p-GSK 3β (Ser9) and enhance the inhibition of Ser9 phosphorylation on GSK 3β kinase activity, thereby activating the plasticity-related CREBeBDNF signaling pathway.

Risk factors for anticoagulant-associated gastrointestinal hemorrhage: a systematic review and meta-analysis

Fuxin Ma,Shuyi Wu,Shiqi Li,Zhiwei Zeng,Jinhua Zhang 대한내과학회 2024 The Korean Journal of Internal Medicine Vol.39 No.1

Background/Aims: There may be many predictors of anticoagulation-related gastrointestinal bleeding (GIB), but until now, systematic reviews and assessments of the certainty of the evidence have not been published. We conducted a systematic review to identify all risk factors for anticoagulant-associated GIB to inform risk prediction in the management of anticoagulation- related GIB. Methods: A systematic review and meta-analysis were conducted to search PubMed, EMBASE, Web of Science, and Cochrane Library databases (from inception through January 21, 2022) using the following search terms: anticoagulants, heparin, warfarin, dabigatran, rivaroxaban, apixaban, DOACs, gastrointestinal hemorrhage, risk factors. According to inclusion and exclusion criteria, studies of risk factors for anticoagulation-related GIB were identified. Risk factors for anticoagulant-associated GIB were used as the outcome index of this review. Results: We included 34 studies in our analysis. For anticoagulant-associated GIB, moderate-certainty evidence showed a probable association with older age, kidney disease, concomitant use of aspirin, concomitant use of the antiplatelet agent, heart failure, myocardial infarction, hematochezia, renal failure, coronary artery disease, helicobacter pylori infection, social risk factors, alcohol use, smoking, anemia, history of sleep apnea, chronic obstructive pulmonary disease, international normalized ratio (INR), obesity et al. Some of these factors are not included in current GIB risk prediction models. such as anemia, co-administration of gemfibrozil, co-administration of verapamil or diltiazem, INR, heart failure, myocardial infarction, etc. Conclusions: The study found that anemia, co-administration of gemfibrozil, co-administration of verapamil or diltiazem, INR, heart failure, myocardial infarction et al. were associated with anticoagulation-related GIB, and these factors were not in the existing prediction models. This study informs risk prediction for anticoagulant-associated GIB, it also informs guidelines for GIB prevention and future research.

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.

Inactive Fusarium Fungal strains (ZSY and MJY) isolation and application for the removal of Pb(II) ions from aqueous environment

Jianyou Long,Gutha Yuvaraja,Shuyi Zhou,Jianying Mo,Huosheng Li,Dinggui Luo,Di Yun Chen,Lingjun Kong,Munagapati Venkata Subbaiah,Guda Mallikarjuna Reddy 한국공업화학회 2019 Journal of Industrial and Engineering Chemistry Vol.72 No.-

In this study, two Fusarium strains namely ZSY and MJY were successfully isolated from the contaminatedsoil of Dabaoshan smelter in Shaoguan City, and utilised as a biosorbent for the removal of Pb(II) ions fromaqueous solution. Effect of pH (6), agitation speed (150 rpm), biomass concentration (1 g/L), initial Pb(II)concentration, contact time (60 min), and effect of temperature (323 K) was investigated. Kinetic andisotherm results showed that the removal of Pb(II) ions onto ZSY and MJY werefitted well with thepseudo-second-order and Langmuir isotherm models (232.56 and 263.16 mg/g). In addition thermodynamicstudies were also discussed.

De novo Assembly and Analysis of Amur Sturgeon (Acipenser schrenckii) Transcriptome in Response to Mycobacterium Marinum Infection to Identify Putative Genes Involved in Immunity

( Qianqian Zhang ),( Xiehao Wang ),( Defeng Zhang ),( Meng Long ),( Zhenbing Wu ),( Yuqing Feng ),( Jingwen Hao ),( Shuyi Wang ),( Qian Liao ),( Aihua Li ) 한국미생물생명공학회(구 한국산업미생물학회) 2019 Journal of microbiology and biotechnology Vol.29 No.8

Fish mycobacteriosis is a common bacterial disease in many species of freshwater and marine fish and has caused severe loss of fish production. Mycobacterium marinum has been the most prevalent pathogen observed in several outbreaks of mycobacteriosis of farmed sturgeons in China. However, the immune responses and pathology of sturgeons in mycobacterial infection are rarely studied. Therefore, we used the Illumina RNA-seq method to analyze the transcriptome profile of Acipenser schrenckii challenged with Mycobacterium marinum. To begin, 168,220 non-redundant contigs were acquired from the infection and control groups, and among these, 33,225 contigs have acquired annotations. A total of 4,043 differently expressed (DE) contigs between the two groups were identified, and among these, 2479 were upregulated and 1564 were down-regulated in the infected fish. A total of 1,340 DE contigs with acquired annotations in KEGG were enriched for 124 pathways including the TNF signaling pathway, and the Toll-like receptor signaling pathway. The roles of DE genes involved in significant pathways and other processes were discussed. The 2,209 DE contigs that have yet to acquire proper annotation may represent candidate genes associated with infection in sturgeons and are expected to serve as immunogenetic resources for further study. To our best knowledge, this is the first transcriptome study on sturgeons under bacterial infection.