BMC PLANT BIOLOGY
·
2021
·Vol. 21
·期 1
·1-12
·
IF 4.215
Wang Yu,Zhang Wenze,Liu Weikang,Ahammed Golam Jalal,Wen Wenxu,Guo Shirong,Shu Sheng,Sun Jin
Background Despite significant limitations of growth medium reuse, a large amount of organic substrate is reused in soilless cultivation of horticultural crops in China. Arbuscular mycorrhizal fungi (AMF) can promote nutrient absorption and improve plant tolerance to biotic and abiotic stresses. However, the mechanisms governing the effects of AMF on crop growth in organic continuous cropping substrates have not been elucidated.Results In this study, we showed that the inoculation of AMF in continuous cropping substrates promoted growth and root development, and increased the root and NADP-malic enzyme (NADP-ME) activity of tomato seedlings. Root transcriptome analysis demonstrated that the plant hormone signal transduction pathway was highly enriched, and 109 genes that positively correlated with the AMF-inoculated plant phenotype were obtained by gene set enrichment analysis (GSEA), which identified 9 genes related to indole acetic acid (IAA). Importantly, the levels of endogenous IAA in tomato seedlings significantly increased after AMF inoculation. Furthermore, the application of AMF significantly increased the expression levels of NADP-ME1 and NADP-ME2, as well as the activity of NADP-ME, and enhanced the root activity of tomato seedlings in comparison to that observed without inoculation of AMF. However, these effects were blocked in plants treated with 2,3,5-triiodobenzoic acid (TIBA), a polar transport inhibitor of IAA.Conclusion sThese results suggest that IAA mediates the AMF-promoted tomato growth and expression of NADP-MEs in continuous cropping substrates. The study provides convincing evidence for the reuse of continuous cropping substrates by adding AMF as an amendment.
研究领域:
植物科学
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农业
·
真菌学
·
园艺学
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产品关联
DOI
PubMed
高影响因子
Biotechnology Journal
·
2025
·Vol. 20
·期 8
·e70090
·
IF 3.1
Hailing Dai,Guoqiang Chen,Jingjiu Mu,Afreen Shagufta,Lijuan Liu,Fan Wang,Sheng Dong,Xiao Men,Lei Wang,Haibo Zhang
Enzymatic biofuel cells face substrate limitations due to enzyme specificity of the electrode. A universal electrode was designed by immobilizing ferredoxin-NADP + oxidoreductase (FNR) with bacterial cellulose (BC), carbon nanotubes (CNTs), and silver nanowires (AgNWs). The electrode coupled with NADPH-dependent malic enzyme or glucose dehydrogenase generated electricity using malic acid and glucose, respectively. The open-circuit voltage reached 79.36 and 75.8 mV, respectively, and the accumulation of pyruvate and gluconate reached 0.30 and 0.25 mM, respectively, after 12 h. This strategy enables electron transfer from diverse substrates via NADPH.
研究领域:
生物燃料电池
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酶催化
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电化学
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纳米材料
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能量转换
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生物技术
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DOI
PubMed
高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
·
2018
·
IF 2.559
Wei Hou,Shiying Lu,Han Zhao,Yan Yu,Haodong Xu,Biao Yu,Lin Su,Chenshui Lin,Benfang Helen Ruan
High levels of glutamate dehydrogenase (GDH) activity are associated with hypoglycemia, cancer, and Parkinson's disease. Propylselen was synthesized to investigate its mechanism of GDH inhibition in comparison with Ebselen and Epigallocatechin gallate (EGCG). Because Ebselen was found to crosslink with the peptide (AA299-341) at the active site of E.coli GDH, the Cys, Pro, and Lys residues of the corresponding peptide were mutagenized to Ala residues. Using enzyme kinetics and biomolecular interaction assays, we found that the conserved GDH P320 residue is important for propylselen binding, C321 for Ebselen binding, and K341 for EGCG binding. In addition, these 3 mutations abolished NADP + binding to E. coli GDH in the absence of glutamate substrate, but in the presence of glutamate, the catalytic activity of the mutants was reduced only by 2–4 fold, indicating that a substrate-induced fit mechanism exists in E. coli GDH. Furthermore, biochemical analysis showed that NADP + had high affinity (Kd of 77 nM) for GDH; by targeting the NADP binding site, propylselen effectively inhibited both E. coli and human GDH activity and improved anticancer activity .
研究领域:
生物化学
·
药理学
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癌症研究
·
酶学
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DOI
PubMed
高影响因子
DRUG METABOLISM AND DISPOSITION
·
2020
·Vol. 48
·期 5
·345-352
·
IF 3.231
Xiaohua Zhao,Hong Ma,Qiusha Pan,Haiyi Wang,Xingkai Qian,Peifang Song,Liwei Zou,Mingqing Mao,Shuyue Xia,Guangbo Ge,Ling Yang
Doxophylline (DOXO) and theophylline are widely used as bronchodilators for treating asthma and chronic obstructive pulmonary disease, and DOXO has a better safety profile than theophylline. How DOXO’s metabolism and disposition affect its antiasthmatic efficacy and safety remains to be explored. In this study, the metabolites of DOXO were characterized. A total of nine metabolites of DOXO were identified in vitro using liver microsomes from human and four other animal species. Among them, six metabolites were reported for the first time. The top three metabolites were theophylline acetaldehyde (M1), theophylline-7-acetic acid (M2), and etophylline (M4). A comparative analysis of DOXO metabolism in human using liver microsomes, S9 fraction, and plasma samples demonstrated the following: 1) The metabolism of DOXO began with a cytochrome P450 (P450)–mediated, rate-limiting step at the C ring and produced M1, the most abundant metabolite in human liver microsomes. However, in human plasma, the M1 production was rather low. 2) M1 was further converted to M2 and M4, the end products of DOXO metabolism in vivo, by non-P450 dismutase in the cytosol. This dismutation process also relied on the ratio of NADP+/NADPH in the cell. These findings for the first time elucidated the metabolic sites and routes of DOXO metabolism in human.
研究领域:
药理学
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生物化学
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产品关联
DOI
PubMed
高影响因子
BIOTECHNOLOGY LETTERS
·
2023
·Vol. 45
·期 8
·993-1000
·
IF 2.7
Peng Shuaiying,Chu Zhongmei,Lu Jianfeng,Li Dongxiao,Wang Yonghong,Yang Shengli,Zhang Yi
The self-sufficient cytochrome P450 BM3 mutant (A74G/F87V/D168H/L188Q) can serve as a biocatalyst for whole-cell catalysis process of indigo. Nevertheless, the bioconversion yield of indigo is generally low under normal cultivation conditions (37 °C, 250 rpm). In this study, a recombinant E. coli BL21(DE3) strain was constructed to co-express the P450 BM3 mutant gene and GroEL/ES genes to investigate whether GroEL/ES can promote the indigo bioconversion yield in E. coli . The results revealed that the GroEL/ES system could significantly increase the indigo bioconversion yield, and the indigo bioconversion yield of the strain co-expressing P450 BM3 mutant and GroEL/ES was about 21-fold that of the strain only expressing the P450 BM3 mutant. In addition, the P450 BM3 enzyme content and in vitro indigo bioconversion yield were determined to explore the underlying mechanism for the improvement of indigo bioconversion yield. The results revealed that GroEL/ES did not increase indigo bioconversion yield by increasing the content of P450 BM3 enzyme and its enzymatic transformation efficiency. Moreover, GroEL/ES could improve the intracellular nicotinamide adenine dinucleotide phosphate (NADPH)/NADP + ratio. Given that NADPH is an important coenzyme in the catalytic process of indigo, the underlying mechanism for the improvement of indigo bioconversion yield is probably related to an increase in the intracellular NADPH/NADP + ratio.
研究领域:
生物化学
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生物技术
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酶学
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产品关联
DOI
PubMed
高影响因子
Synthetic and Systems Biotechnology
·
2025
·
IF 4.4
Haizhao Xue,Yinghan Hu,Aabid Manzoor Shah,Xueying Wang,Xiaojia Guo,Yanzhe Huang,Zongbao K. Zhao
The nicotinamide cofactors including nicotinamide adenine dinucleotide (NAD) and its phosphate (NADP) play important roles in facilitating redox reactions for energy metabolism and biosynthesis. To expand the cofactor menu, a non-natural cofactor nicotinamide cytosine dinucleotide (NCD) has been introduced recently. The reduced forms of these cofactors carry reducing equivalents that are essential for cellular metabolism. However, there is a long standing challenge to rationally transfer reducing equivalent from one cofactor to another, albeit such process is highly demanding in metabolic engineering. This study develops a new approach based on malic enzyme (ME)-mediated transhydrogenation to enable reducing equivalents exchange among different cofactors. We used wild-type ME, MaeB and an engineered ME* that favors NAD, NADP and NCD, respectively, to demonstrate such conversions. When an in vitro system initiated with equal amount of NADH and NCD in the presence of ME, ME* and excess amount of pyruvate was held for 2 h, up to 65% NADH was consumed and 57% NCDH was generated. When implemented into NCD self-sufficient Escherichia coli cells, the system directed reducing equivalents toward NCDH-linked formation of lactate. Overall, this work offers an effective strategy to regulate intracellular reducing equivalents that may serve as a novel tool for metabolic engineering and synthetic biology.
研究领域:
生物化学
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分子生物学
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代谢工程
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合成生物学
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微生物学
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DOI
PubMed
高影响因子
Cell Death & Disease
·
2024
·Vol. 15
·期 10
·1-16
·
IF 8.1
Wan Xinyi,Zeng Weiwu,Fan Haonan,Wang Chenliang,Han Shixun,Sun Zhongxing,Tang Mei,Shao Juejia,Liu Yu,Fang Yuan,Jia Junqi,Tang Yin,Zhang Yanjun,Zhao Bin,Fang Dong
MAT2B works together with MAT2A to synthesize S-Adenosyl methionine (SAM) as the primary methyl donor. MAT2B, despite lacking catalytic activity, exerts regulatory control over the enzymatic activity of MAT2A. In addition to the enzymatic activity regulation, we find that, in an NADP + -dependent manner, MAT2B binds and stabilizes MAT2A. Disruption of the cellular NADP + remodels the protein level of MAT2A. The pentose phosphatase pathway regulates the level of MAT2A protein through the interaction of NADP + with MAT2B. Additionally, MAT2B-MAT2A interaction regulates the mRNA m6A modification and stability. In liver tumors, the Mat2a mRNA level is elevated but the protein level is decreased by the restricted NADP + . Blocking the interaction between MAT2B and MAT2A by the keto diet can suppress liver tumor growth. These findings reveal that MAT2B is essential for regulating the protein levels of MAT2A and connecting SAM synthesis to mRNA m6A.
研究领域:
癌症生物学
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分子生物学
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生物化学
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产品关联
DOI
PubMed
高影响因子
ACS Synthetic Biology
·
2025
·
IF 3.9
Xinzhe Zhang,Aipeng Li,Xiaohan Huang,Shuqi Guo,Chenyue Zhang,Ramon Gonzalez,Qiang Fei
The growing demand for natural products in pharmaceutical applications has prompted a focus on more sustainable methods to produce high-value terpenoids using microbial cell factories. Due to the characteristics of high abundance, renewable, and low cost, methane has emerged as a promising feedstock for biomanufacturing. In this study, the methanotrophic bacterium Methylotuvimicrobium buryatenese 5GB1C, known for its industrial potential, was metabolically engineered to synthesize taxadiene, a crucial precursor in paclitaxel production. The biosynthesis of taxadiene from methane was first established by employing an endogenous strong promoter to enhance the expression of heterologous taxadiene synthase in M. buryatenese 5GB1C. To further optimize the metabolic flux, rate-limiting enzymes (Dxs and IspA) were upregulated in the methylerythritol phosphate pathway while complementing the native pathway with the essential idi gene that was originally deficient. Coupled with a 2.1-fold improvement in the NADPH/NADP+ ratio, these modifications collectively boosted taxadiene production from 2.58 to 22.97 mg/L in serum vials. Ultimately, a temperature-controlled two-stage cultivation was implemented in 3 L bioreactors, which achieved a remarkable titer of 104.88 mg/L, representing the highest reported titer for diterpenoid biosynthesis from methane. This work demonstrates the potential of utilizing methane for the sustainable production of advanced terpenoids with reduced environmental impact.
研究领域:
生物技术
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微生物学
·
合成生物学
·
生物工程
·
工业生物技术
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产品关联
DOI
PubMed
高影响因子
Advanced Science
·
2025
·
IF 14.1
Heng Du,Xu Liu,Jianghua Shen,Hailong Yuan,Hao Zhang,Gan Xi,Yujing Li,Yuhan Wang,Jiahe Zhang,Chaofan Yang,Pengfei Xu,Jiawan Wang,Fang Wang,Siqi Liu,Yanan Zhou,Qi Gu,Jingjing Lu,Tuo Wei,Zeyu Gao,Jingyi Zang,Jun Wang,Moshi Song
Myocardial ischemia/reperfusion (I/R) injury is a major contributor to myocardial damage, leading to adverse cardiac remodeling and dysfunction. Recent studies have highlighted the potential of gut microbiota-derived metabolites in modulating cardiac outcomes. Here, the cardioprotective effects of a commensal bacterium Flavonifractor plautii ( F. plautii ) and its metabolite desaminotyrosine (DAT) against myocardial I/R injury are investigated. We showed that prophylactic gavage of F. plautii attenuates myocardial I/R injury as evidenced by improved cardiac function and reduced cardiac injury. We also found that its metabolite DAT recapitulates these cardioprotective effects against myocardial I/R injury. Transcriptomic analysis has revealed that DAT preserves cardiac tissue and attenuates immune responses against myocardial I/R injury. Mechanistically, DAT promotes cardiomyocyte survival through the modulation of the nicotinamide adenine dinucleotide phosphate (NADP + /NADPH) ratio. Further, DAT suppressed macrophage proinflammatory activities and cardiac inflammation via the reduction in interleukin-6 (IL-6) production. Taken together, our findings indicate that F. plautii and its metabolite DAT exert pleiotropic cardioprotective effects against myocardial I/R injury, suggesting them as potential prophylactic therapeutic options for alleviating myocardial I/R injury.
研究领域:
心脏病学
·
微生物学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2024
·
IF 3.4
Xinqi Xu,Yaping Meng,Bingmei Su,Juan Lin
Esomeprazole is the most popular proton pump inhibitor for treating gastroesophageal reflux disease. Previously, a phenylacetone monooxygenase mutant LnPAMOmu15 (LM15) was obtained by protein engineering for asymmetric synthesis of esomeprazole using pyrmetazole as substrate. To scale up the whole cell asymmetric synthesis of esomeprazole and reduce the cost, in this work, an Escherichia coli whole-cell catalyst harboring LM15 and formate dehydrogenase from Burkholderia stabilis 15516 ( Bst FDH) were constructed through optimized gene assembly patterns. CRISPR/Cas9 mediated insertion of P trc promoter in genome was done to enhance the expression of key genes to increase the cellular NADP supply in the whole cell catalyst, by which the amount of externally added NADP + for the asymmetric synthesis of esomeprazole decreased to 0.05 mM from 0.3 mM for reducing the cost. After the optimization of reaction conditions in the reactor, the scalable synthesis of esomeprazole was performed using the efficient LM15- Bst FDH whole-cell as catalyst, which showed the highest reported space-time yield of 3.28 g/L/h with 50 mM of pyrmetazole loading. Isolation procedure was conducted to obtain esomeprazole sodium of 99.55 % purity and > 99.9 % ee with 90.1 % isolation yield. This work provides the basis for production of enantio-pure esomeprazole via cost-effective whole cell biocatalysis.
研究领域:
生物催化
·
药物化学
·
生物技术
·
代谢工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Sustainable Chemistry & Engineering
·
2019
·
IF 6.97
Yan Zhang,Yin-Qi Wu,Na Xu,Qian Zhao,Hui-Lei Yu,Jian-He Xu
Enzymatic asymmetric sulfoxidation using molecular oxygen as the oxidant is a promising green chemistry approach to chiral sulfoxide production. Despite the broad substrate spectrum of cyclohexanone monooxygenases (CHMOs), some unnatural substrates with bulky functional groups, such as the pharmaceutically relevant omeprazole sulfide, cannot be effectively accepted by CHMOs. Herein, we describe a set of variants derived from an Acinetobacter calcoaceticus CHMO (AcCHMO), whose active sites adjacent to the substrate tunnel were altered to shift the substrate specificity from cyclohexanone monooxygenation toward omeprazole sulfide sulfoxidation. We performed homologous modeling and molecular docking to identify key residues that might affect the substrate specificity. Two libraries of residues lining the active center of AcCHMO were then constructed and screened by an effective halo-based selection method using the solubility difference between the substrate (omeprazole sulfide) and product (esomeprazole). Functional evaluation of the resultant variants showed that the substrate specificity of AcCHMO was markedly altered from the small natural substrate (cyclohexanone) toward the desired bulky substrate (omeprazole sulfide) despite the extremely poor activity detected even for the best variant, M2 (0.61 U/gprot). The crystal structure of M2 complexed with a flavin adenine dinucleotide (FAD) prosthetic group was determined, which provided insight into the altered substrate specificity. To improve the activity of enzyme M2 toward pharmaceutical precursor omeprazole sulfide, we performed both local and global protein engineering among the two CASTing libraries surrounding FAD+ and NADP+ prosthetic groups and an error-prone PCR library of the full-length AcCHMO. As a result, variant M6 was obtained, giving a 50-fold higher activity compared to M2. This structure-guided protein engineering of AcCHMO provided a promising candidate for converting omeprazole sulfide into (S)-omeprazole using a green biocatalytic method.
研究领域:
生物催化
·
酶工程
·
药物化学
·
绿色化学
·
蛋白质结构与功能
✓ 已验证
DOI
高影响因子
Frontiers in Microbiology
·
2021
·
IF 5.64
Li Jingxin,Zhang Yuxiao,Wang Xing,Walk Seth T.,Wang Gejiao
Antimony (Sb)-resistant bacteria have potential applications in the remediation of Sb-contaminated sites. However, the effect of Sb(III) exposure on whole-cell metabolic change has not been studied. Herein, we combined untargeted metabolomics with a previous proteomics dataset and confirmatory gene transcription analysis to identify metabolic responses to Sb(III) exposure in Agrobacterium tumefaciens GW4. Dynamic changes in metabolism between control and Sb(III)-exposed groups were clearly shown. KEGG pathway analysis suggested that with Sb(III) exposure: (1) the branching pathway of gluconeogenesis is down-regulated, resulting in the up-regulation of pentose phosphate pathway to provide precursors of anabolism and NADPH; (2) glycerophospholipid and arachidonic acid metabolisms are down-regulated, resulting in more acetyl-CoA entry into the TCA cycle and increased capacity to produce energy and macromolecular synthesis; (3) nucleotide and fatty acid synthesis pathways are all increased perhaps to protect cells from DNA and lipid peroxidation; (4) nicotinate metabolism increases which likely leads to increased production of co-enzymes (e.g., NAD+ and NADP+) for the maintenance of cellular redox and Sb(III) oxidation. Expectedly, the total NADP+/NADPH content, total glutathione, and reduced glutathione contents were all increased after Sb(III) exposure in strain GW4, which contribute to maintaining the reduced state of the cytoplasm. Our results provide novel information regarding global bacterial responses to Sb(III) exposure from a single gene level to the entire metabolome and provide specific hypotheses regarding the metabolic change to be addressed in future research.
研究领域:
微生物学
·
环境科学
·
分子生物学
·
代谢组学
✓ 已验证
DOI
PubMed
高影响因子
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