Engineering
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2025
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IF 11.6
Wenyue Yan,Baogang Zhang,Yi’na Li,Jianping Lu,Yangmei Fei,Shungui Zhou,Hailiang Dong,Fang Huang
Microbial vanadate (V(V)) reduction is a key process for environmental geochemistry and detoxification of vanadium (V). However, the electron transfer pathways and V isotope fractionation involved in this process are not yet fully understood. In this study, the V(V) reduction mechanisms with concomitant V isotope fractionation by the gram-positive bacterium Bacillus subtilis ( B. subtilis ) and the gram-negative bacterium Thauera humireducens ( T. humireducens ) were investigated. Both strains could effectively reduce V(V), and (90.5% ± 1.6%) and (93.0% ± 1.8%) of V(V) at an initial concentration of 50 mg·L −1 was removed over 10 d incubation, respectively. V(V) was bioreduced to insoluble vanadium (IV), which was distributed both inside and outside the cells. Electron transfer via cytochrome C, nicotinamide adenine dinucleotide, and glutathione played critical roles in V(V) reduction. Metabolomic analysis showed that differentially enriched metabolites (quinone, biotin, and riboflavin) mediated electron transfer in both strains. The aqueous V in the remaining solution became isotopically heavier as V(V) bioreduction proceeded. The obtained V isotope composition dynamics followed a Rayleigh fractionation model, and the isotope enrichment factor ( ε ) was (–0.54‰ ± 0.04‰) for B. subtilis and (–0.32‰ ± 0.03‰) for T. humireducens , with an insignificant difference. This study provides molecular insights into electron transfer for V(V) bioreduction and reveals V isotope fractionation during this bioprocess, which is helpful for understanding V biogeochemistry and developing novel strategies for V remediation.
研究领域:
环境微生物学
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地球化学
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生物化学
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DOI
高影响因子
MICROCHIMICA ACTA
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2022
·Vol. 190
·期 1
·1-11
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IF 6.408
Xu Yixuan,Shi Xuerong,Ran Fanpeng,Zhang Ziqi,Phipps Josh,Liu Xiaoyan,Zhang Haixia
A novel Tb-doped Eu-based metal–organic framework (Eu-MOF@Tb) has been developed by incorporating hexanuclear europium cluster and 2,2′-bipyridine-5,5′-dicarboxylic acid as well as coordination with Tb(III). Owing to the diverse coordination status of Tb(III) and Eu(III) in MOF, antenna effect emission from Tb(III) can be invoked by dipicolinic acid (DPA), but the luminescence originating from Eu(III) remains unchanged. Taking advantage of this phenomenon, a ratiometric luminescent method for detection of DPA, a biomarker for Bacillus subtilis spores, was developed through differential sensitization toward lanthanide ions. This analysis method allowed for the detection of DPA in the 0.2–10 μM concentration range, with a detection limit of 60 nM. It was further validated by spiked recoveries (89.3–110%) of real-world samples with RSD values in the range 3.9–11%. Alongside this, a paper indicator test was prepared for naked-eye detection of DPA via a dose-sensitive color evolution from red to green under UV light. The effectiveness of the proposed approach was explored in the detection of bacterial spores in real biological and environmental samples and indicated great potential for applications as a real-time monitoring system against the anthrax threat. Graphical Abstract
研究领域:
分析化学
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生物化学
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环境科学
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DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
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2013
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IF 2.823
Long-Hua Zhou,Na Wang,Wei Zhang,Zong-Bo Xie,Xiao-Qi Yu
An facile and green one-pot route has been developed for the synthesis of chromenes using salicylaldehyde and α,β-unsaturated ketones. α-Amylase from Bacillus subtilis shows excellent catalytic activity and exerts good adaptability to different substrates in the reaction. This promiscuous enzyme-catalyzed domino reaction not only extends the application of α-amylase from B. subtilis for new chemical transformations, but also provided an alternative synthetic method for 2 H -chromene derivatives.
研究领域:
生物化学
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有机合成
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酶学
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DOI
高影响因子
iScience
·
2023
·
IF 5.8
Qilin Yu,Haohao Mao,Bowen Yang,Yahui Zhu,Cheng Sun,Zhiqiang Zhao,Yang Li,Yaobin Zhang
Electrical stimulation has been used to strengthen microbial extracellular electron transfer (EET), however, the deep-seated reasons remain unclear. Here we reported that Bacillus subtilis , a typical gram-positive bacterium capable of extracellular respiration, obtained a higher EET capacity after the electrical domestication. After the electrical domestication, the current generated by the EET of B. subtilis was 23.4-fold that of the control group without pre-domestication. Multiple lines of evidence in bacterial cells of B. subtilis , their cell walls, and a model tripeptide indicated that the polarization of amide groups after the electrical stimulation forwarded the H-bonds recombination and radical generation of protein-like substances to develop extracellular electron transfer via the proton-coupled pattern. The improved electrochemical properties of protein-like substances benefited the trans -cell-wall electron transfer and strengthen extracellular respiration. This study was the first exploration to promote microbial extracellular respiration by improving the electrochemical properties of protein-like substances in cell envelopes.
研究领域:
微生物学
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生物物理学
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电化学
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DOI
高影响因子
Cell Reports
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2022
·
IF 9.995
Kaining Zhang,Shanshan Li,Yawen Wang,Zhihao Wang,Nancy Mulvenna,Hang Yang,Peipei Zhang,Huan Chen,Yan Li,Hongliang Wang,Yongxiang Gao,Sivaramesh Wigneshweraraj,Steve Matthews,Kaiming Zhang,Bing Liu
Summary Enolase is a highly conserved enzyme that presents in all organisms capable of glycolysis or fermentation. Its immediate product phosphoenolpyruvate is essential for other important processes like peptidoglycan synthesis and the phosphotransferase system in bacteria. Therefore, enolase inhibitors are of great interest. Here, we report that Gp60, a phage-encoded enolase inhibitor protein (PEIP) of bacteriophage SPO1 for Bacillus subtilis , is an enolase inhibitor. PEIP-expressing bacteria exhibit growth attenuation, thinner cell walls, and safranin color in Gram staining owing to impaired peptidoglycan synthesis. We solve the structure of PEIP-enolase tetramer and show that PEIP disassembles enolase by disrupting the basic dimer unit. The structure reveals that PEIP does not compete for substrate binding but induces a cascade of conformational changes that limit accessibility to the enolase catalytic site. This phage-inspired disassembly of enolase represents an alternative strategy for the development of anti-microbial drugs.
研究领域:
微生物学
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分子生物学
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结构生物学
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生物化学
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药学
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DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2019
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IF 3.371
Zhengyu Tao,Gang Fu,Sijia Wang,Zhaoxia Jin,Jianping Wen,Dawei Zhang
The wider application of creatinase, an enzyme used to determine renal function, is limited by the yield problem and current production cannot meet the needs of society. Therefore, the work constructed an efficient B. subtilis expression strain for creatinase production based on cell leakage. Initial expression of creatinase in B. subtilis was achieved using the native promoter P hpa Ⅱ . Subsequently, the promoter was optimized, and the efficient expression of creatinase was realized by using the maltose-induced promoter P glv , which increased the enzyme activity 5-fold and the properties of creatinase were analyzed. Meanwhile, it was found that creatinase without a signal peptide could be secreted into the extracellular medium, so the secretion of creatinase in B. subtilis was investigated. Disabling the classical secretory pathways didn’t have an effect on the expression level of extracellular creatinase, indicating that the excretion of creatinase didn’t depend on these pathways. Calcein-AM/PI double staining results showed that the membrane of the expression strain was damaged, while electron microscopy images revealed that leakage sites were present on the surface of the expressing bacteria, which led to the passive leakage of creatinase from B. subtilis . Finally, the enzyme activity was further improved to 141.9 U/mL in 5-L high-density fermentation.
研究领域:
微生物学
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生物技术
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酶学
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发酵技术
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DOI
高影响因子
JOURNAL OF POWER SOURCES
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2025
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IF 7.9
Sunday O. Oguntomi,Obinna M. Ajunwa,Cinzia Calvio,Pellegrino Conte,Massimiliano Galluzzi,Sze Shin Low,Simone Morra,Run Pan,Dexin Wang,Enrico Marsili
Gene expression in electroactive bacteria (EAB), which sense electrochemical stimuli and elicit genetic response, can be regulated through electrogenetic methods. Although differential gene expression patterns have been observed in biofilms grown on polarized electrodes, little is known about the molecular mechanisms that sense electrochemical potential and bring about the genetic regulation in response to this potential. Here, we observe that the molecular weight of poly gamma glutamic acid (γ-PGA) produced in Bacillus subtilis PB5760 biofilms changed with the applied electrochemical potential. The application of 0.2 and 0.4 V vs. Ag/AgCl reduced the molecular weight of γ-PGA in the biofilm from 3800 to 4070 kDa at open circuit potential (OCP) to 1040–1590 kDa and 2170–2730 kDa, respectively. Under the same conditions, the relative levels of expression of pgdS , a gene that codes for a γ-PGA-specific endo-hydrolase in cells growing at OCP, 0.2 and 0.4 V vs. Ag/AgCl were 1.12 ± 0.57, 2.74 ± 0.54, 1.61 ± 0.57, respectively, which correlates with the molecular weight reduction. This is the first biotechnological application of electrogenetics for producing γ-PGA with a tunable molecular weight. Results show the feasibility of electrogenetics approach for gene regulation in EAB.
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DOI
高影响因子
JOURNAL OF BIOTECHNOLOGY
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2021
·
IF 3.307
Bo Gao,Ning Zhao,Jieying Deng,Yang Gu,Shiru Jia,Ying Hou,Xueqin Lv,Long Liu
Methanol is a promising green feedstock for producing fuels and chemicals because it is inexpensive, clean, environmentally friendly, and easily prepared. Thus, many studies have been devoted to engineering non-native methylotrophic platform microorganisms to utilize methanol. This study adopted a series of strategies to develop a synthetic methylotrophic Bacillus subtilis that can use methanol as the carbon source, including the heterologous expression of methanol dehydrogenase (Mdh), enhancement of the expressions of 3-hexulose-6-phosphate synthase (Hps) and 6-phospho-3-hexuloisomerase (Phi), regulation of the expressions of key enzymes at both the translational and transcriptional levels, stabilization of the key enzyme expression through a dual-system for expressing the target genes on both the plasmid and genome, and improvement of the catalytic activity of Mdh with a recycling strategy for NAD + . As a result, the methanol consumption of the synthetic methylotrophic B. subtilis reached 4.09 g/L, with the maximum OD 600 showing a 2.21-fold increase compared with the wild-type B. subtilis , which cannot use methanol. We further deleted the phosphoglucose isomerase (Pgi) and added co-substrates to increase the supply of ribulose-5-phosphate (Ru-5-P), and the specific methanol consumption rate increased by an additional 27.54%. Finally, we successfully constructed two strains that cannot grow in M9 medium with xylose or ribose unless methanol is utilized. The strategies used in this study are generally applicable to other studies on synthetic methylotrophy .
研究领域:
微生物学
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代谢工程
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合成生物学
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产品关联
DOI
PubMed
高影响因子
FOOD CHEMISTRY
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2022
·
IF 9.231
Hao Luo,Yihong Bao,Ping Zhu
Functional lycopene-rich yogurt displays attractive nutritious and health-promoting benefits among existing functional dairy products, owing to supplement with lycopene which could enhance immunity, prevent cancer, and cardiovascular diseases. Due to poor stability and fat-solubility of lycopene, its incorporation into yogurt is challengeable. In this study, carotenoid genes for lycopene synthesis were co-introduced into probiotic Bacillus subtilis for efficient lycopene production. Further engineered B. subtilis was applied as adjunct starter culture for achieving lycopene-rich yogurt. Developed yogurt exhibited desirable physiochemical characteristics compared with plain yogurt. Moreover, lycopene-rich yogurt was endowed with significantly high antioxidant capacity. More importantly, this functionalized yogurt had attractive sensorial attributes for quality-assured food to facilitate consumer acceptance. As the first report of fortifying yogurt of lycopene using B. subtilis with improved functional properties, this study offers a new and facile clue to enrich bioactive lycopene and probiotic B. subtilis in yogurt for healthy and nutritional food development.
研究领域:
食品科学
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微生物学
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营养学
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2017
·
IF 3.154
Chen Hsu,Bo-Yuan Wu,Yu-Chuan Chang,Chi-Fon Chang,Tai-Ying Chiou,Nan-Wei Su
The soy isoflavones daidzein (DAI) and genistein (GEN) have beneficial effects on human health. However, their oral bioavailability is hampered by their low aqueous solubility. Our previous study revealed two water-soluble phosphorylated conjugates of isoflavones, daidzein 7-O-phosphate and genistein 7-O-phosphate, generated via biotransformation by Bacillus subtilis BCRC80517 cultivated with isoflavones. In this study, two novel derivatives of isoflavones, daidzein 4′-O-phosphate and genistein 4′-O-phosphate, were identified by HPLC-ESI–MS/MS and 1H, 13C, and 31P NMR, and their biotransformation roadmaps were proposed. Primarily, isoflavone glucosides were deglycosylated and then phosphorylated predominantly into 7-O-phosphate conjugates with traces of 4′-O-phosphate conjugates. Inevitably, trace quantities of glucosides were converted into 6″-O-succinyl glucosides. GEN was more efficiently phosphorylated than DAI. Nevertheless, the presence of GEN prolonged the time until the exponential phase of cell growth, whereas the other isoflavones showed little effect on cell growth. Our findings provide new insights into the novel microbial phosphorylation of isoflavones involved in xenobiotic metabolism.
研究领域:
微生物学
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生物化学
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药学
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代谢学
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天然产物化学
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DOI
高影响因子
iScience
·
2025
·
IF 4.1
Shuai Liu,Yi Li,Lin Quan,Hai-Xia Liu,Yang Luo,Yong-Zhong Wang
SUMMARY Optimizing cellulase biosynthesis in B. subtilis is crucial for enhancing enzymatic yield in lignocellulosic biomass conversion. However, the regulatory mechanisms linking intracellular NAD(H/ + ) levels to cellulase production remain elusive. In this study, we systematically screened thirteen genes associated with NAD + biosynthesis and NADH regeneration in B. subtilis Z2. Employing a modular engineering strategy with four distinct modules, we directed metabolic flux to enhance NAD + biosynthesis and NADH regeneration. Key genes ( ycel , nadV , nadM , mdh , and sucB ) were identified, and their co-expression in B. subtilis Z2 resulted in a 13.09-fold increase in intracellular NADH levels and a consequential 2.24- and 2.04-fold enhancement in the filter paper-hydrolyzing (FPase [representing total cellullase]) activity and carboxymethylcellulase (CMCase [representing endoglucanase]) activity, respectively. Experimental validations, including antagonist LaCl 3 treatment and spcF gene deletion, unequivocally established the calcium signaling pathway's role in regulating cellulase gene overexpression in response to elevated intracellular NAD(H/ + ) levels.
研究领域:
微生物学
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生物化学
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合成生物学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
·
2015
·
IF 3.082
Lei Rao,Zhenzhen Xu,Yongtao Wang,Feng Zhao,Xiaosong Hu,Xiaojun Liao
The objective of this study was to investigate the inactivation of the Bacillus subtilis spores by high pressure CO 2 combined with high temperature (HPCD + HT) and to analyze the clumping effect of the spores on their HPCD + HT resistance. The spores of B . subtilis were subjected to heat at 0.1 MPa and HPCD at 6.5–25 MPa, and 82 °C, 86 °C, and 91 °C for 0–120 min. The spores were effectively inactivated by HPCD + HT, but a protective effect on the spores was also found, which was closely correlated to the pressure, temperature and time. The spores treated by HPCD + HT at 6.5 and 10 MPa exhibited a two-stage inactivation curve of shoulder and log-linear regions whereas the spores at 15–25 MPa exhibited a three-stage inactivation curve of shoulder, log-linear and tailing regions, and these curves were well fitted to the Geeraerd model. Approximately 90% of pyridine-2,6-dicarboxylic acid (DPA) was released after HPCD + HT and the 90% DPA release time depend on the pressure and temperature. Moreover, the spore clumping in suspensions was examined by dynamic light scattering. The particle size of the spore suspensions increased with the increase of pressure, temperature and time, indicating the spore clumping. 0.1% Tween 80 as a surfactant inhibited the spore clumping and increased the inactivation ratio of the spores by HPCD + HT. These results indicated that the spore clumping enhanced the spores' resistance to HPCD + HT and induced a protective effect.
研究领域:
微生物学
·
食品科学
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产品关联
DOI
PubMed
高影响因子
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