BIOMEDICAL MICRODEVICES
·
2020
·Vol. 22
·期 4
·1-8
·
IF 2.176
Li Xiao,Zhang Man,Hu Yujie,Xu Jian,Sun Dongke,Hu Tao,Ni Zhonghua
Rapid on-site detection of glucose has been attracting considerable attention nowadays. Screen-printed electrodes (SPEs) were assessed as ideal detection platforms due to their advantages such as, disposability, portability, ease of miniaturization, and mass production. The topology and shape of electrodes have a crucial impact on their electrical conductivity and electrochemical properties. In this study, a versatile electrochemical platform with optimized three-electrode configuration was developed through screen-printing technology. Three types of SPEs were prepared, and their electrocatalytic properties toward glucose detection were investigated. Based on this platform, both enzyme-based (denoted as GOD/rGO) and non-enzymatic (denoted as Co@MoS 2 /rGO) bioactive compounds were deposited on the working electrode of the circular SPE through simply drop-casting, respectively. Their morphology was characterized through scanning electron microscopy (SEM). Cycle sweep voltammetry was used to study the electrocatalytic activity of these biosensors. The circular SPE exhibited satisfying electrochemical redox activity and much higher sensitivity towards glucose detection, which rendered it a promising candidate for point-of-care detection.
研究领域:
生物医学工程
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分析化学
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产品关联
DOI
PubMed
高影响因子
CHINESE SCIENCE BULLETIN
·
2013
·Vol. 58
·期 26
·3266-3273
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IF 1.319
Wu Qiong,Bai LinQuan,Liu WeiCheng,Li YingYing,Lu CaiGe,Li YaQian,Lin ZhenYa,Wang Meng,Xue ChunSheng,Chen Jie
Streptomyces lydicus A01 resists many plant pathogens (including Fusarium spp.) by producing the antifungal agent natamycin, which binds to the ergosterol of fungal cell membranes and inhibits the growth of pathogens. Trichoderma harzianum CECT2413 is a widely-distributed soil fungus that antagonizes several plant fungal pathogens (including Fusarium spp.) by producing chitinase and degrading chitin, a major component of the fungal cell wall. This study attempted to enhance the biocontrol effect of S. lydicus A01 on Fusarium spp. by transforming the chitinase gene of Trichoderma . Chitinase and natamycin could act synergistically on both the cell walls and cell membranes of pathogens. The 33-kD chitinase-encoding gene ( chit33 ) was cloned and conjugal-transformed from T. harzianum CECT2413 to S. lydicus A01, and then confirmed via polymerase chain reaction (PCR) assays. Subsequent analyses using the 3,5-dinitrosalicylic acid (DNS) method and ultraviolet spectrophotometry showed that compared with its wild type strain (WT), the S. lydicus A01 conjugal transformant (CT) with chit33 gene exhibited substantially higher chitinase activity and natamycin production. The resistance of S. lydicus A01- chit33 CT and WT to four Fusaria in crops and vegetables was tested via the cup-plate method. Compared with the WT, the conjugal transformant of S. lydicus A01 with chit33 gene from T. harzianum CECT2413 showed greatly increased biocontrol effect on fusarium disease. This study would be beneficial to the development of high-quality antifungal bio-agents for agricultural applications via the synergy between the previously non-existent and pre-existing functions achieved through heterogeneous gene transformation.
研究领域:
微生物学
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植物病理学
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分子生物学
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DOI
高影响因子
Chilean Journal of Agricultural Research
·
2017
·Vol. 77
·期 3
·257-265
·
IF 0.719
Wei Li,Lele Zhang,Ning Zhao,Wenlei Guo,Weitang Liu,Lingxu Li,Jinxin Wang
ABSTRACT American slough grass ( Beckmannia syzigachne (Steud.( Fernald) is a worldwide weed and is widely distributed in rice-wheat rotations in China. Fenoxaprop- P -ethyl and mesosulfuron-methyl are two major herbicides used to control B. syzigachne . Resistance has evolved in B. syzigachne under continuous selective pressure from herbicides. This study aimed to establish the cross-resistance pattern of a resistant population and explore the potential non-target-site based resistance mechanisms of B. syzigachne . Sequencing of target enzyme genes (acetyl coenzyme A carboxylase (ACCase( and acetolactate synthase (ALS() revealed that there were no resistance-endowing amino acid substitutions in the resistant B. syzigachne population (R F1 ) compared with the sensitive population (S F1 ), and obtained the purified materials. Furthermore, piperonyl butoxide (PBO) and malathion showed synergistic effects with fenoxaprop- P -ethyl and mesosulfuron-methyl respectively in R F1 . Therefore, we speculate that the resistance observed in B. syzigachne was related with metabolic, mostly involving the cytochrome P450 enzymes. Cross resistance patterns showed that the purified resistant B. syzigachne produce high resistance to fenoxaprop- P -ethyl and pyroxsulam; intermediate resistance to flucarbazone-sodium; low resistance to quizalofop- P -ethyl, clodinafop-propargyl, sethoxydim and mesosulfuron-methyl; sensitive to clethodim pinoxaden and isoproturon. Key words: Herbicide resistance; metabolic resistance; P450 inhibitors; target enzymes sequencing.
研究领域:
植物学
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分子生物学
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农业科学
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产品关联
DOI
PubMed
高影响因子
ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY
·
2021
·Vol. 108
·期 2
·e21835
·
IF 1.698
Mao-Ye Li,Yun Wang,Xiao Lei,Chuan-Tao Xu,Dong-Dong Wang,Su Liu,Shi-Guang Li
The green peach aphid, Myzus persicae (Sulzer), is a serious agricultural pest with a worldwide distribution. Catalase (CAT), which is encoded by the catalase ( Cat ) gene, is an extremely important antioxidant enzyme that plays a pivotal role in protecting cells against the toxic effects of hydrogen peroxide. The Cat gene has not been characterized in M. persicae ; therefore, this study describes the identification of the Cat ( MpCat ) gene from M. persicae . MpCat contains an open reading frame of 1515 bp and encodes a MpCAT protein consisting of 504 amino-acid residues. MpCAT possesses features typical of other insect catalases, including 7 conserved amino acids involved in binding heme and 15 involved in binding nicotinamide adenine dinucleotide phosphate. Phylogenetic analysis showed that MpCAT was closely related to orthologs from other aphid species. MpCat consisted of nine exons and eight introns, and the number and insertion sites of introns are consistent with those of Cat genes from Acyrthosiphon pisum (Harris) and Aphis gossypii Glover. The mRNA transcripts of MpCat were detected at all tested developmental stages, with the highest mRNA level in alate adults. The expression of MpCat was significantly upregulated when M. persicae was exposed to low and high temperatures, ultraviolet radiation, Beauveria bassiana , and permethrin. The transcription of MpCat and the activity of catalase were suppressed by RNA interference, and knockdown of MpCat significantly reduced the survival rate in M. persicae under heat stress. The results provide valuable information for further study on the physiological functions of MpCat . Research Highlights A catalase gene ( MpCat ) was characterized in Myzus persicae . Transcription of MpCat was responsive to various environmental stresses. Knockdown of MpCat increased mortality in M. persicae under heat stress.
研究领域:
昆虫学
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分子生物学
·
遗传学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2024
·
IF 6.2
Wenjun Xiong,Wei Wei,Man He,Bin Hu,Jun Men,Jiawei Tu,Wei Miao
Biomethylation is an effective means of arsenic detoxification by organisms living in aquatic environments. Ciliated protozoa (including Tetrahymena species) play an important role in the biochemical cycles of aquatic ecosystems and have a potential application in arsenic biotransformation. This study compared arsenic tolerance, accumulation, methylation, and efflux in 11 Tetrahymena species. Nineteen arsenite (As(III)) S -adenosylmethionine (SAM) methyltransferase ( arsM ) genes, of which 12 are new discoveries, were identified, and protein sequences were studied. We then constructed recombinant cell lines based on the Tetrahymena thermophila ( T. thermophila ) wild-type SB210 strain and expressed each of the 19 arsM genes under the control of the metal-responsive the MTT1 promoter. In the presence of Cd 2+ and As(V), expression of the arsM genes in the recombinant cell lines was much higher than in the donor species. Evaluation of the recombinant cell line identified one with ultra-high arsenic methylation enzyme activity, significantly higher arsenic methylation capacity and much faster methylation rate than other reported arsenic methylated organisms, which methylated 89% of arsenic within 6.5 h. It also had an excellent capacity for the arsenic detoxification of lake water containing As(V), 56% of arsenic was methylated at 250 μg/L As(V) in 48 h. This study has made a significant contribution to our knowledge on arsenic metabolism in protozoa and demonstrates the great potential to use Tetrahymena species in the arsenic biotransformation of aquatic environments.
研究领域:
环境生物学
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分子生物学
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生态学
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产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
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2020
·
IF 2.952
Xuelian Zhou,Yi Shen,Yong Xu,Venkatesh Balan
2-Keto- d -gluconic acid (2-KGA) is a high-value-added product from lignocellulosic straw-based biorefining process obtained via cell catalysis using Gluconobacter oxydans ( G. oxydans ); nonetheless, the economical production has some limitations because of the formation of undesirable mixture products of keto-gluconic acid from glucose. In this study, it was demonstrated that pulse addition of hydrogen peroxide (H 2 O 2 ) to synthetic media was beneficial to improve the conversion rate of glucose to 2-KGA by 1.47-fold in G. oxydans . Moreover, some lignocellulosic degradation compounds served as substitute and played the role of H 2 O 2 on corn stover hydrolysate for the production of 2-KGA. By conducting qRT-PCR analysis, eight dehydrogenase genes were identified in G. oxydans , which were responsible for regulating 2-KGA production. Moreover, most of the identified genes were found to produce membrane-bound glucose dehydrogenase and gluconate 2-dehydrogenase, and were up-regulated by adding H 2 O 2 or degradation compounds. These up-regulated genes could be responsible for directing cell catalysis of glucose to 2-KGA in G. oxydans .
研究领域:
生物化学
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微生物学
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产品关联
DOI
高影响因子
PEST MANAGEMENT SCIENCE
·
2021
·Vol. 77
·期 6
·2737-2746
·
IF 4.845
Mei-Mei Wang,Gui-Jun Long,Huan Guo,Xuan-Zheng Liu,Hong Wang,Youssef Dewer,Zhao-Qun Li,Kun Liu,Qiu-Liang Zhang,Yun-Feng Ma,Peng He,Ming He
BACKGROUND Carboxyl/cholinesterases (CCEs) are thought to play a pivotal role in the degradation of sex pheromones and plant-derived odorants in insects, but their exact biochemistry and physiological functions remain unclear. RESULTS In this study, two paralogous antennae-enriched CCEs from Plutella xylostella (PxylCCE16a and 16c) were identified and functionally characterized. High-purity protein preparations of active recombinant PxylCCE16a and 16c have been obtained from Sf 9 insect cells by Ni 2+ affinity purification. Our results revealed that the purified recombinant PxylCCE016c is able to degrade two sex pheromone components Z9-14:Ac and Z11-16:Ac at 27.64 ± 0.79% and 24.40 ± 3.07%, respectively, while PxylCCE016a presented relatively lower activity. Additionally, a similar difference in activity was measured in plant-derived odorants. Furthermore, both CCEs displayed obvious preferences for the two sex pheromone components, especially on Z11-16:Ac ( K m values are in the range 7.82–45.06 μmol L –1 ) which much lower than plant odorants ( K m values are in the range 1290–4030 μmol L –1 ). Furthermore, the activity of the two newly identified CCEs is pH-dependent. The activity at pH 6.5 is obviously higher than that at pH 5.0. Interestingly, only PxylCCE016c can be inhibited by a common esterase inhibitor triphenyl phosphate (TPP) with LC 50 of 1570 ± 520 μmol L –1 . CONCLUSION PxylCCE16c plays a more essential role in odorant degradation than PxylCCE16a. Moreover, the current study provides novel potential pesticide targets for the notorious moth Plutella xylostella . © 2021 Society of Chemical Industry
研究领域:
昆虫学
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生物化学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
·
2023
·
IF 4.4
Meng Zhang,QingYan Li,Feiyu Fan,Fuping Lu,XueLi Zhang
This study highlights an effective strategy based on the synergistic complementarity of metabolic engineering and adaptive evolution.First, the chassis strain YL013 was obtained by deleting five acyl-CoA oxidase genes to reduce the degradation of γ-decalactone(GDL). Then, adaptive laboratory evolution with increasing concentrations of γ-decalactone was employed to obtain the strain YL013–250 with increased γ-decalactone tolerance of up to 250 mg/L. Moreover, this strain produced 664.5 mg/L γ-decalactone, representing a 142 % improvement over the parental strain YL013. Furthermore, four candidate fatty acyl-CoA synthetase genes, FAA1 , FAA2 FAA3 and FAT1, were modulated using the strong, constitutive TEFin promoter to investigate their roles in GDL production. The results showed that overexpression of FAA1 , FAA2 and FAA3 can effectively improve the ability of Yarrowia lipolytica to synthesize GDL using ricinoleic acid as substrate, among which overexpression of FAA1 had the most significant effect on GDL yield. Finally, gene of alcohol acyltransferase ( AAT1 ) from Prunus persica was introduced into Y. lipolytica , leading to a GDL titer of 2.2 g/L, which was a 1.32-fold increase compared with the parental strain without AAT1 . Taken together, this study provides a combinatorial strategy to improve GDL production through rational regulation of pathway genes and non-rational adaptive laboratory evolution.
研究领域:
代谢工程
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微生物生物技术
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生物过程工程
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生物化学
✓ 已验证
产品关联
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2014
·
IF 5.9
Zhaohao Wang,Xi Luo,Qijin Wan,Kangbing Wu,Nianjun Yang
A versatile matrix was fabricated and utilized as a universal interface for the construction of enzyme-based biosensors. This matrix was formed on the gold electrode via combining self-assembled monolayer of 2,3-dimercaptosuccinic acid with gold nanoparticles. Gold nanoparticles were electrochemically deposited. Electrochemistry of three redox enzymes (catalase, glucose oxidase, and horseradish peroxidase) was investigated on such a matrix. The electrocatalytic monitoring of hydrogen peroxide and glucose was conducted on this matrix after being coated with those enzymes. On them the monitoring of hydrogen peroxide and glucose shows rapid response times, wide linear working ranges, low detection limits, and high enzymatic affinities. This matrix is thus a versatile and suitable platform to develop highly sensitive enzyme-based biosensors.
研究领域:
生物化学
·
纳米技术
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF CULTURAL HERITAGE
·
2022
·
IF 3.229
Juwen Guo,Encong Shi,Xinxin Mu,Jing Han,Wenxuan Qu,Yan Song,Sang Da,Ozhu Dawa,Feng Gao,Ling Guo,Austin Nevin
This work focuses on the study of the removal of overpaint from the stone carvings of the Potala Palace, Tibet. The removal of historical mortar and overpaint is a significant challenge for the conservation of stone carvings and requires careful optimization. Paints applied in the past were made of a mixture of hydraulic lime, water and milk, and are causing damage to the carvings of the Potala Palace. The protein content of lime-based paint samples collected from the Potala Palace was determined by enzyme-linked immunosorbent assay (ELISA). To optimize the removal of the overpaint without causing abrasion or other damage to the underlying stone carvings, we prepared a simulated paint with the same protein content. Removal rate and color difference were used to select the best cleaning agent from ionic surfactant, ethanol/aqueous solutions, five proteases and Twin-20 (non-ionic surfactant). With color difference and gloss as evaluation indexes, the influencing factors (temperature, pH, enzyme concentration, and enzyme hydrolysis time) were measured. Results suggest that alkaline protease was the most suitable cleaning agent for removing surface paint, and the best working condition is an enzymatic dosage of 1.92 mg/mL under 42 °C, pH of 9.5, with a hydrolysis time of 5.4 min. This method generated the desired color difference of 2.25 and the minimal gloss (0.1 ± 0.1 GU) on the underlying stone, with no significant variation from the predicted value ( P < 0.05). Before and after cleaning, the change in surface roughness was less than 10%. This study provided a feasibility plan for removing the paints from the outer wall of the Potala Palace by using enzymatic hydrolysis.
研究领域:
文化遗产保护
·
化学
·
材料科学
✓ 已验证
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Yi Xu,Guoxiang You,Shihong Yang,Jie Zhang,Jun Hou
Nano zero-valent iron (nZVI) assisted anaerobic digestion (AD) is considered an effective approach for mediating antibiotic resistance genes (ARGs) dissemination in activated sludge. However, little is known about the mechanisms that drive antibiotic resistomes evolution in relation with their physiological roles distinct from protection of bacteria against antimicrobials. Metagenomic sequencing showed that efflux pumps and rRNA methyltransferase genes were the most prevalent ARGs in anaerobes, being responsible for the variations of antibiotic resistome upon nZVI addition. Particularly, to withstand the oxidative stimuli derived from initially 9 days nZVI exposure, genes involved in the two component systems ( PhoRB , BaeSR , EvgSA ) and global regulator networks ( MarA , SoxS ) were increased, activating the chromosomally encoded intrinsic efflux pumps expression. Furthermore, to benefit for survival, the polyspecific efflux pumps were involved in the microbial physiological adaptations of enhanced siderophores, lipid moieties, and quorum sensing molecules exportation, which induced the adaptative efflux pumps expression. On the other hand, to maintain the protein synthesis under oxidative condition, genes encoding Rsm, Erm, and Rlu family methyltransferases were notably up-regulated via horizontal transfer, inducing the acquired rRNA methylation resistance. Importantly, after prolonged exposure to nZVI for 18 and 27 days, the antibiotic resistomes were measurably decreased, due to the recovered oxidative state and increased membrane rigidity. Overall, these findings provide a deep understanding on the necessity of antibiotic resistome evolution for anaerobes to adapt to nZVI exposure for survival, which can be introduced to improve the ARGs risks analysis.
研究领域:
环境科学
·
微生物学
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分子生物学
✓ 已验证
DOI
高影响因子
ACS ES&T Engineering
·
2024
·
IF 7.4
Yong Li,Bohan Chen,Xiaolei Zhang,Zhizhan Luo,Mengen Lei,Tao Song,Youcheng Wei,Ji Li,Zhiyun Long,Jun Ma
Elemental sulfur autotrophic partial denitrification (S0-PDN) can supply nitrite for anaerobic ammonium oxidation without an organic carbon requirement. This study aimed to evaluate the impact of high salinity on S0-PDN and investigate the feasibility of an enhanced strategy involving the addition of glycine betaine (GB). The results showed that high salinity inhibited S0-PDN by over 60% as salinity increased from low (0 and 5 g/L) to high (10 and 15 g/L) levels, even though Thiobacillus (65.9%–84.0%) was always the dominant genus. The reason might be the suppression of high salinity on the bioactivity. Surprisingly, the addition of GB (1 mmol/L) effectively enhanced the performance of S0-PDN under high salinity conditions (10 g/L), resulting in a 5.1-fold increase in the NO3–-N conversion efficiency and a 3.1-fold increase in the effluent NO2–-N concentration. Metagenomics analysis revealed that high salinity did not alter the dominant microbial composition. GB facilitated the recovery of the nitrate conversion capacity of sulfur autotrophic denitrifying bacteria with the up-regulation of nitrate reductase genes (by 18.1%) and Sox genes (by 21.4%). However, GB also stimulated the growth of heterotrophic denitrifying bacteria. Part of the nitrite produced in S0-PDN was reduced through heterotrophic denitrification with GB as an electron donor. Overall, the addition of GB was demonstrated as an effective approach to mitigate the inhibitory effects of high salinity on S0-PDN, which offers valuable theoretical support for its application in high-salinity wastewater treatment.
研究领域:
环境工程
·
微生物学
✓ 已验证
DOI
高影响因子
危险品化学品经营许可证（带存储）