BIORESOURCE TECHNOLOGY
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2024
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IF 9.7
Jing Wang,Min Du,Xin Wang,Junchen He,Alei Zhang,Kequan Chen
To achieve industrial-scale putrescine production, a high efficient bio-synthesis of putrescine involving arginase (ARG, EC 3.5.3.1) and L-ornithine decarboxylase was evaluated here. Among the four arginases tested, ARG BT from Bos Taurus showed the highest activity (1966 U/mg). Compared to the L-arginine decarboxylase (ADC) pathway, the strain expressing ARG BT and L-ornithine decarboxylase (SpeC) produced 28.7 g/L putrescine, a 38.6 % increase. Two pyridoxal phosphate (PLP) salvage pathways were evaluated, and the strain BL-P Tac -PdxK co-expressed pyridoxal kinase (PdxK) performed better. D-Glucose was used as the co-substrate to improve the putrescine titer further. Under optimal conditions, 43.6 g/L putrescine was produced from 87.1 g/L L-arginine, and 76 g/L putrescine was synthesized on a 0.5 L scale. Using L-arginine fermentation broth (60 g/L) as the substrate, a titer of 30 g/L putrescine was achieved. This efficient biotransformation process presented here enables feasible industrial-scale putrescine production.
研究领域:
生物工程
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微生物学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2021
·Vol. 413
·期 15
·3925-3932
·
IF 4.157
Li Shuangqin,Fu Guoqing,Wang Yaya,Xiang Yueci,Mu Shuai,Xu Yixuan,Liu Xiaoyan,Zhang Haixia
Acid phosphatase has become a significant indicator of prognostic and medical diagnosis, and its dysfunction may lead to a series of diseases. A novel dual-signal fluorescence method for acid phosphatase detection based on europium polymer (europium-pyridine dicarboxylicacid-adenine) and pyridoxal phosphate (PLP) was proposed. PLP coordinated with europium polymer via Eu 3+ and P–O bonds, and the fluorescence of europium polymer was quenched due to the photoinduced electron transfer (PET) effect between aldehyde and europium polymer. Upon addition of acid phosphatase, the PLP was transformed to phosphate (Pi) and pyridoxal (PL). The PL was released from the surface of europium polymer, and the blue emission was enhanced due to the formation of internal hemiacetal, while the fluorescence of europium polymer recovered. The blue (PL) and red emission (Eu 3+ ) were positively correlated with acid phosphatase activity; thus the sensitive assay of acid phosphatase was effectively achieved. The two signals were applied to determine the acid phosphatase with limits of detection (LOD) of 0.04 mU/mL and 0.38 mU/mL, and the linear ranges were 0.13–5.00 mU/mL and 1.25–20.00 mU/mL, respectively. The probe can be used to trace the acid phosphatase in biological systems and holds promise for use in clinical diagnosis and early prevention. Graphical abstract
研究领域:
生物化学
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医学诊断
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DOI
PubMed
高影响因子
Communications Biology
·
2025
·Vol. 8
·期 1
·1-14
·
IF 5.1
Xu Tianqi,Fang Dan,Li Fulei,Wang Zhiqiang,Liu Yuan
Antimicrobial resistance poses a severe threat to human health, with colistin serving as a critical medication in clinical trials against multidrug-resistant Gram-negative bacteria. However, the efficacy of colistin is increasingly compromised due to the rise of MCR-positive bacteria worldwide. Here, we reveal a notable metabolic disparity between mcr -positive and -negative bacteria through transcriptome and metabolomics analysis. Specifically, pyridoxal 5’-phosphate (PLP), the active form of vitamin B6, was significantly diminished in mcr -positive bacteria. Conversely, supplementing with PLP could reverse the metabolic profile of drug-resistant bacteria and effectively restore colistin’s bactericidal properties. Mechanistically, PLP was found to augment bacterial proton motive force by inhibiting the Kdp transport system, a bacterial K + transport ATPase, thereby facilitating the binding of the positively charged colistin to the negatively charged bacterial membrane components. Furthermore, PLP supplementation triggers ferroptosis-like death by accumulating ferrous ions and inducing lipid peroxidation. These two modes of action collectively resensitize mcr -harboring Gram-negative bacteria to colistin therapy. Altogether, our study provides a novel metabolic-driven antibiotic sensitization strategy to tackle antibiotic resistance and identifies a potentially safe antibiotic synergist.
研究领域:
微生物学
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药理学
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代谢组学
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DOI
PubMed
高影响因子
ChemistrySelect
·
2022
·Vol. 7
·期 45
·e202203721
·
IF 2.307
Weihua Yu,Jianhua Kuang,Qingqing Hu,Zhonghua Wang,Yunwen Liao,Zhengjun Cheng
Graphical A new ratio fluorescent probe (RF-probe) based on the mixing of red-emitting D-penicillamine (DPA) stabilized copper nanoclusters (DPA-CuNCs) with pyridoxal 5’-phosphate (PLP) was designed for sensitive and selective determination of Al 3+ . The excess accumulation of nonessential Al 3+ in the human body can cause serious health problems, thus there is significant interest in the detection of Al 3+ . A new ratio fluorescent probe (RF-probe) based on the mixing of red-emitting D-penicillamine (DPA) stabilized copper nanoclusters (DPA-CuNCs) with pyridoxal 5’-phosphate (PLP) was designed for sensitive and selective determination of Al 3+ . The −CHO group of PLP is expected to form an imine linkage with the free −NH 2 group available of the DPA on the surface of DPA-CuNCs. The content of Al 3+ could be mensurated by the as-prepared RF-probe with a linear response in the range of 1.33–46.67 μmol ⋅ L −1 , and corresponding limit of detection (LOD) was calculated to be 0.33 μmol ⋅ L −1 . Moreover, the RF-probe was successfully extended to the assay of Al 3+ in aluminum carbonate chewing tablets, baking powder, pond water and drink water with satisfactory results compared to the standard atomic absorption spectroscopy (AAS).
研究领域:
分析化学
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环境科学
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生物医学
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DOI
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2025
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IF 3.8
Yan Wang,Bo Ding,Qingqing Bao,Yunying Zhao,Guohui Li,Yu Deng
Agmatine is an important basic drug and nutritional supplements, which exhibits significant physiological activities. Traditional methods, such as whole-cell catalysis, require substantial pyridoxal 5′-phosphate (PLP) addition, which accounts for 15 % of production costs. In this study, we utilized purified arginine decarboxylase (AdiA-M2) that leverages the intrinsic PLP cofactor, without the need for additional PLP supplementation, and ZIF-8 was employed to enhance the pH tolerance. The successful incorporation of AdiA-M2 into the ZIF-8 structure was confirmed through scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The immobilized arginine decarboxylase demonstrated markedly enhanced performance under alkaline conditions, along with improved thermal stability and storage activity compared to its free enzyme counterpart. Specifically, the immobilized AdiA@ZIF-8 system achieved an initial conversion rate of 95.16 %, after five recycles the titer attained 266.69 g/L without the extra PLP addition, which is 1.67 times higher than the best current production approach. The adopted enzyme-catalysis system eliminated the need for additional cofactors, thereby lowering costs and providing valuable insights for the industrial production of agmatine.
研究领域:
生物化学
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酶工程
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制药工程
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DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Jing Wang,Xin Xu,Wanqing Wei,Wei Song,Jian Wen,Guipeng Hu,Xiaomin Li,Cong Gao,Xiulai Chen,Liming Liu,Jing Wu
Pyridoxal 5′-phosphate (PLP) is highly valuable in food and medicine. However, achieving the efficient biosynthesis of PLP remains challenging. Here, a salvage pathway using acid phosphatase from Salmonella typhi (StAPase) and pyridoxine oxidase from Escherichia coli (EcPNPO) as pathway enzymes was established for the first time to synthesize PLP from pyridoxine (PN) and pyrophosphate (PPi). StAPase was identified as a rate-limiting enzyme. Two protein modification strategies were developed based on the PN phosphorylation mechanism: (1) improving the binding of PN into StAPase and (2) enhancing the hydrophobicity of StAPase’s substrate binding pocket. The kcat/Km of optimal mutant M7 was 4.9 times higher than that of the wild type. The detailed mechanism of performance improvement was analyzed. Under the catalysis of M7 and EcPNPO, a PLP high-yielding strain of 14.5 ± 0.55 g/L was engineered with a productivity of 1.0 ± 0.02 g/(L h) (the highest to date). The study suggests a promising method for industrial-scale PLP production.
研究领域:
生物化学
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合成生物学
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代谢工程
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DOI
PubMed
高影响因子
INSECT MOLECULAR BIOLOGY
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2024
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IF 2.3
Hui-Hua Cao,Wei-Wei Kong,Xi-Ya Chen,Sadaf Ayaz,Cai-Ping Hou,Yi-Sheng Wang,Shi-Huo Liu,Jia-ping Xu
The RNA interference pathway mediated by microRNAs (miRNAs) is one of the methods to defend against viruses in insects. Recent studies showed that miRNAs participate in viral infection by binding to target genes to regulate their expression. Here, we found that the Bombyx mori miRNA, miR-6498-5p was down-regulated, whereas its predicted target gene pyridoxal phosphate phosphatase PHOSPHO2 ( BmPLPP2 ) was up-regulated upon Bombyx mori nucleopolyhedrovirus (BmNPV) infection. Both in vivo and in vitro experiments showed that miR-6498-5p targets BmPLPP2 and suppresses its expression. Furthermore, we found miR-6498-5p inhibits BmNPV genomic DNA (gDNA) replication, whereas BmPLPP2 promotes BmNPV gDNA replication. As a pyridoxal phosphate (PLP) phosphatase (PLPP), the overexpression of BmPLPP2 results in a reduction of PLP content, whereas the knockdown of BmPLPP2 leads to an increase in PLP content. In addition, exogenous PLP suppresses the replication of BmNPV gDNA; in contrast, the PLP inhibitor 4-deoxypyridoxine facilitates BmNPV gDNA replication. Taken together, we concluded that miR-6498-5p has a potential anti-BmNPV role by down-regulating BmPLPP2 to modulate PLP content, but BmNPV induces miR-6498-5p down-regulation to promote its proliferation. Our findings provide valuable insights into the role of host miRNA in B. mori –BmNPV interaction. Furthermore, the identification of the antiviral molecule PLP offers a novel perspective on strategies for preventing and managing viral infection in sericulture.
研究领域:
分子生物学
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病毒学
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昆虫学
·
RNA干扰
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DOI
PubMed
高影响因子
PHARMACOLOGY
·
2019
·
IF 1.615
Chen Xuyang,Li Hui,Luo Hongjun,Lin Zhexuan,Luo Wenhong
Background/Aims: Hydrazone and acylhydrazone derivatives, which are produced from aldehyde reacting with hydrazine or acylhydrazine, have been reported to exhibit antitumor activities. However, the angionenic effects of this kind of derivatives haven’t been elucidated. Here, we synthesized 12 pyridoxal hydrazone and acylhydrazone compounds and investigated their antiangiogenic effects and the underlying mechanisms. Method: 3-(4,5-Dimethylthiazol-2-yl)-2, 5-dipheyltetrazolium bromide assay was used to screen the inhibitory effects of the synthesized compounds on endothelial cells (ECs) proliferation. The compound with best inhibitory effect was further evaluated with wound-healing assay and tube formation assay. Calcein-Am assay was carried out to determine the content of intracellular labile iron pool (LIP). Intracellular reduced glutathione (GSH) was determined by spectrophotometry. Flow cytometry was used to determine cell cycle and apoptosis. Results: Compound 10 (3-hydroxy-5-[hydroxymethyl]-2-methyl-pyridine-4-carbaldehyde-2-naphthalen-1-acetyl hydrazone) showed the best inhibitory effect on human umbilical vascular ECs proliferation, with IC50 value of 25.4 μmol/L. It not only inhibited wound-healing and tube formation of ECs, but also decreased the content of intracellular LIP and GSH. Furthermore, it arrested ECs cycle at S phase and induced cell apoptosis. Conclusions: Compound 10 exhibits antiangiogenic effects by reducing the content of intracellular LIP and GSH, and subsequently arresting cell cycle and inducing cell apoptosis
研究领域:
药理学
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肿瘤学
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分子生物学
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DOI
PubMed
高影响因子
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
·
2022
·Vol. 414
·期 17
·4877-4884
·
IF 4.478
Deng Hao-Hua,Yang Hui-Jing,Huang Kai-Yuan,Zheng Yi-Jing,Xu Ying-Ying,Peng Hua-Ping,Liu Yin-Huan,Chen Wei,Hong Guo-Lin
As a kind of sensing and imaging fluorescent probe with the merit of low toxicity, good stability, and environment-friendly, silicon nanoparticles (SiNPs) are currently attracting extensive research. In this work, we obtained mitoxantrone-SiNPs (MXT-SiNPs) with green emission by one-pot synthesis under mild temperature condition. The antenna based on pyridoxal phosphate (PLP) was designed for light-harvesting to enhance the luminescence of MXT-SiNPs and to establish a novel sensing strategy for alkaline phosphatase (ALP). PLP transfers the absorbed photon energy to MXT-SiNPs by forming Schiff base. When PLP is dephosphorized by ALP, the released free hydroxyl group reacts with aldehyde group to form internal hemiacetal, which leads to the failure of Schiff base formation. Based on the relationship between antenna formation ability and PLP hydrolysis degree, the activity of ALP can be measured. A good linear relationship was obtained from 0.2 to 3.0 U/L, with a limit of detection of 0.06 U/L. Furthermore, the sensing platform was successfully used to detect ALP in human serum with recovery of 97.6–106.2%. The rational design of antenna elements for fluorescent nanomaterials can not only provide a new pathway to manipulate the luminescence, but also provide a new direction for fluorescence sensing strategy.
研究领域:
生物化学
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医学检测
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纳米材料科学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2021
·
IF 6.953
Min Zou,Jing-Yan Li,Meng-Jia Zhang,Jun-Hui Li,Jun-Tao Huang,Pei-Dan You,Shu-Wen Liu,Chun-Qiong Zhou
At present, there are still no anti-Zika virus (ZIKV) drugs or vaccines approved by FDA with accurate targets and antiviral mechanisms. Considering the RNA G-quadruplex sequences in ZIKV genome, it is very meaningful to develop G-quadruplex binders as potential anti-ZIKV drugs with novel and accurate targets. In this paper, five classical G-quadruplex binders including Ber , Braco 19 , NiL , 360A and PDS have been chosen to discuss their interaction with ZIKV RNA G-quadruplexes. PDS shows higher binding affinity and thermal stability than the other G-quadruplex binders. This property is further evidenced in cells by immunofluorescence microscopy. And PDS shows higher anti-ZIKV activity (EC 50 = 4.2 ± 0.4 μM) than the other G-quadruplex binders as well as the positive control ribavirin, with a low cytotoxicity. By time-of-addition assay, PDS exerts antiviral activity at the post-entry process of ZIKV replication cycle, thus inhibiting ZIKV mRNA replication and protein expression. Furthermore, PDS combines with ZIKV NS2B-NS3 protease and reduces its catalytic activity. This study suggests that G-quadruplex binder PDS is an effective multi-target ZIKV inhibitor, which provides more guidance to design some novel anti-ZIKV drugs targeting ZIKV RNA G-quadruplexes.
研究领域:
病毒学
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药物化学
·
分子生物学
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DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2020
·
IF 3.448
Wenlong Zheng,Kaitong Chen,Sai Fang,Xiuli Cheng,Gang Xu,Lirong Yang,Jianping Wu
A number of organic synthesis involve threonine aldolase (TA), a pyridoxal phosphate (PLP)-dependent enzyme. Although the addition of exogenous PLP is necessary for the reactions, it increases the cost and complicates the purification of the product. This work constructed a PLP self-sufficient biocatalysis system for TA, which included an improvement of the intracellular PLP level and co-immobilization of TA with PLP. Engineered strain BL-ST was constructed by introducing PLP synthase PdxS/T to Escherichia coli BL21(ED3). The intracellular PLP concentration of the strain increased approximately fivefold to 48.5 μmol/g DCW . l -TA, from Bacillus nealsonii ( Bn LTA), was co-expressed in the strain BL-ST with PdxS/T, resulting in the engineered strain BL- Bn LTA-ST. Compared with the control strain BL- Bn LTA (254.1 U/L), the enzyme activity of the strain BL- Bn LTA-ST reached 1518.4 U/L without the addition of exogenous PLP. An efficient co-immobilization system was then designed. The epoxy resin LX-1000HFA wrapped by polyethyleneimine (PEI) acted as a carrier to immobilize the crude enzyme solution of the strain BL- Bn LTA-ST mixed with an extra 100 μM of exogenous PLP, resulting in the catalyst HFA PEI - Bn LTA-STPLP 100. HFA PEI - Bn LTA-STPLP 100 exhibited a half-life of approximately 450 h, and the application of the catalyst in the continuous biosynthesis of 3-[4-(methylsulfonyl) phenyl] serine had more than 180 batch reactions (>60% conv ) without the extra addition of exogenous PLP. The excellent compatibility and stability of the system were further confirmed by other TAs. This work introduced a PLP self-sufficient biocatalysis system that can reduce the cost of PLP and contribute to the industrial application of TA. In addition, the system may also be applied in other PLP-dependent enzymes.
研究领域:
生物化学
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酶工程
·
工业微生物学
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DOI
PubMed
高影响因子
JOURNAL OF CHROMATOGRAPHY A
·
2017
·
IF 3.981
Qian Wang,Xiao-Mei He,Xi Chen,Gang-Tian Zhu,Ren-Qi Wang,Yu-Qi Feng
Phosphorylation is a crucial post-translational modification, which plays pivotal roles in various biological processes. Analysis of phosphopeptides by mass spectrometry (MS) is intractable on account of their low stoichiometry and the ion suppression from non-phosphopeptides. Thus, enrichment of phosphopeptides before MS analysis is indispensable. In this work, we employed pyridoxal 5′-phosphate (PLP), as an immobilized metal affinity chromatography (IMAC) ligand for the enrichment of phosphopeptides. PLP was grafted onto several substrates such as silica (SiO 2 ), oxidized carbon nanotube (OCNT) and silica coated magnetic nanoparticles (Fe 3 O 4 @SiO 2 ). Then the metal ions Fe 3+ , Ga 3+ and Ti 4+ were incorporated for the selective enrichment of phosphopeptides. It is indicated that Fe 3 O 4 @SiO 2 -PLP-Ti 4+ has a superior selectivity towards phosphopeptides under as much as 1000-fold interferences of non-phosphopeptides. Further, Fe 3 O 4 @SiO 2 -PLP-Ti 4+ exhibited high efficiency in selective enrichments of phosphopeptides from complex biological samples, including human serum and tryptic digested non-fat milk. Finally, Fe 3 O 4 @SiO 2 -PLP-Ti 4+ was successfully employed in the sample pretreatment for profiling phosphopeptides in a tryptic digest of rat brain proteins. Our experimental results evidenced a great potential of this new chelator-based material in phosphoproteomics study.
研究领域:
生物化学
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蛋白质组学
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分析化学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）