CHEMISTRY & BIODIVERSITY
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2022
·Vol. 19
·期 9
·e202200118
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IF 2.745
Bing-Kang Sun,Rui-Yu Wang,Bei Li,Xing Fan,Yuan Zhou,Bing Gu,Yang-Yang Yan
A rapid and accurate analytical method was established to identify CREC and CSEC. Orbitrap-MS was used to detect the polypeptide of CREC and CSEC strains, and MS data were analyzed by pattern recognition analyses such as hierarchical cluster analysis (HCA), principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and orthogonal partial least squares discriminant analysis (OPLS-DA). HCA based on the farthest distance method could well distinguish the two types of E. coli , and the cophenetic correlation coefficient of the farthest distance method was 0.901. Comparing the results of PCA, PLS-DA, and OPLS-DA, OPLS-DA exhibited the highest accuracy in predicting the CREC and CSEC strains. A total of 26 compounds were identified, and six of the compounds were the highly significant difference between the two types of strains. MS combined with pattern recognition can achieve a more comprehensive and efficient statistical analysis of complex biological samples. Graphical
研究领域:
微生物学
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分析化学
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PubMed
高影响因子
BIOMEDICINE & PHARMACOTHERAPY
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2022
·
IF 7.419
Jingyi Hu,Yiheng Tong,Zhaofeng Shen,Yanan Li,Cheng Cheng,Ryan Au,Feng Xu,Yajun Liu,Lei Zhu,Hong Shen
Ulcerative colitis (UC) is a chronic and relapsing inflammatory disease of the intestine. Dysbiosis, especially the expansion of facultative anaerobic Enterobacteriaceae , maybe the main pathogenesis of UC. Gegen Qinlian decoction (GD), a traditional Chinese medicinal formula chronicled in the Shang Han Lun , is commonly used to treat UC and has shown an excellent effect on inducing disease remission. However, the role of GD in regulating gut microbiota has not been fully clarified. Herein, we investigated the potential effect of GD on inhibiting the expansion of Enterobacteriaceae and further explored the potential mechanism of this action. Our study demonstrated that GD remarkably reduced body weight loss of colitis mice, shortening of colon length, and inflammation of the colon. Peroxisome proliferator-activated receptor-γ (PPAR-γ) signaling was inactivated in colitis colon tissue, and the abundance of Escherichia coli ( E. coli, family of Enterobacteriaceae ) in colonic contents and the concentration of lipopolysaccharide (LPS) in colonic tissue were significantly upregulated after DSS-treatment. Notably, GD administration can result in the activation of PPAR-γ and inactivation of iNOS, which lead to the reduction of nitrate, the inhibition of E. coli, and less production of LPS. Combined GD with PPAR-γ antagonist, the effect of GD on the treatment of UC was weakened, and effectless in inhibiting the expansion of Enterobacteriaceae . Therefore, GD ameliorates UC by preventing a dysbiotic expansion of potentially pathogenic E. coli by reducing nitrate levels in the lumen through activating PPAR-γ signaling.
研究领域:
药理学
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微生物学
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免疫学
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胃肠病学
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PubMed
高影响因子
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
·
2018
·Vol. 84
·期 20
·
IF 3.633
Haiquan Yang,Xiao Lu,Jinyuan Hu,Yuan Chen,Wei Shen,Long Liu
Escherichia coli is one of the most widely used host microorganisms for recombinant protein expression and metabolic engineering, but it cannot efficiently secrete recombinant proteins to extracellular space. Here, extracellular protein secretion was enhanced in E. coli by deleting two d,d-carboxypeptidase genes (dacA and dacB, single and double deletions) to perturb the cell wall peptidoglycan network. Deletion of dacA and dacB enhanced the accumulation of intracellular soluble peptidoglycan in E. coli and affected cell morphology, resulting in a more irregular cell shape and the appearance of transparent bulges. Deletion of dacA and dacB appears to disrupt the normal rigid structure, presumably due to perturbation and destruction of the cell wall peptidoglycan network. The extracellular green fluorescent protein (GFP) fluorescence intensity of deletion mutants was increased by >2.0-fold compared with that of control cells, and that of the double deletion mutant was increased by 2.7-fold. Extracellular recombinant fibroblast growth factor receptor 2 (FGFR2) and collagen E4 secretion in deletion mutants was also enhanced compared with that in the control cells. Additionally, the extracellular recombinant amylase activity of single-deletion mutants BL21 ΔdacA pETDuet-amyk and BL21 ΔdacB pETDuet-amyk was increased 2.5- and 3.1-fold, respectively. The extracellular distribution of α-galactosidase by deletion mutants was also increased by >2.0-fold. Deletion of dacA and dacB increased outer membrane permeability, which could explain the enhanced extracellular protein secretion.IMPORTANCE Cell surface structure stabilization is important for extracellular secretion of proteins in Escherichia coli. As the main constituent of the cell wall, peptidoglycan contributes to cell structure robustness and stability. Here, we perturbed the peptidoglycan network by deleting dacA and dacB genes encoding d,d-carboxypeptidase enzymes to improve extracellular protein secretion. This new strategy could enhance the capacity of E. coli as a microbial cell factory for extracellular secretion of proteins and chemicals.
研究领域:
生物工程
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微生物学
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PubMed
高影响因子
Synthetic and Systems Biotechnology
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2025
·
IF 4.4
Xinru Wang,Lian Wang,Qihang Chen,Ke Wang,Huijing Wang,Dong Li,Song Gao,Weizhu Zeng,Jingwen Zhou
Hydroxysalidroside is an important natural phenylethanoid glycoside with broad application prospects in the food and pharmaceutical fields. However, its low concentration in plants and complex extraction hinder its production. Despite being a promising way to synthesize hydroxysalidroside in Escherichia coli , glycosylation remains the limiting factor for its production. A de novo biosynthetic pathway for hydroxysalidroside was successfully constructed in E. coli via the screening of glycosyltransferase, overexpressing phosphoglucomutase ( pgm ) and UDP-glucose pyrophosphorylase ( galU ) to ensure a sufficient supply of UDP-glucose (UDPG). Additionally, a semi-rational design of UGT85A1 was conducted to expand the acceptor-binding pocket to eliminate steric hindrance interfering with the binding of hydroxytyrosol. The endogenous genes ushA and otsA were knocked out to further reduce the consumption of UDPG. Finally, a titer of 5837.2 mg/L was achieved in a 5 L fermenter by optimizing the feeding times of carbon sources. This laid the foundation for the subsequent biosynthesis of phenylethanoid glycosides.
研究领域:
生物化学
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合成生物学
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PubMed
高影响因子
BIOORGANIC CHEMISTRY
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2025
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IF 4.7
Jianianhua Luo,Jiahui Xu,Qi Li,Lulu Wu,Jianjun Pei,Linguo Zhao
Indican, abundant in plant stems and leaves, serves as a precursor for indigo and indirubin, possessing application potential in many industries. In this study, a multi-faceted experimental strategy for producing indican utilizing tryptophan as the starting material in E. coli was established. This system incorporated the synergistic action of TnaA, MeFMO, and PtUGT2 M91A/S145A , a mutant with a 20 % increase in activity of glycosyltransferase PtUGT2, all co-expressed within the bacterial host. Both a UDP-glucose recycling system and an NADPH regeneration system were incorporated for donor regeneration. Finally, a co-culture system aimed at reducing metabolic burdens was designed. After optimization, a maximum yield of 1043 mg/L was achieved, with a molar conversion rate of 48.1 %, representing an almost 14-fold improvement over the performance of a single strain. This study offers a plant-independent synthesis process for indican, paving a new path for year-round indican production.
研究领域:
生物工程
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代谢工程
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PubMed
高影响因子
mLife
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2025
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IF 4.5
Huahui Chen,Fengfeng Zheng,Xi Feng,Zijing Huang,Wei Yang,Chuanlun Zhang,Wenbin Du,Kira S. Makarova,Eugene V. Koonin,Zhirui Zeng
Eukaryotes are hypothesized to be archaeal–bacterial chimeras. Given the different chemical structures of membrane phospholipids in archaea and bacteria, transformations of membranes during eukaryogenesis that led to the bacterial-type membranes of eukaryotic cells remain a major conundrum. One of the possible intermediates of eukaryogenesis could involve an archaeal–bacterial hybrid membrane. So far, organisms with hybrid membranes have not been discovered, and experimentation on such membranes has been limited. To generate mixed membranes, we reconstructed the archaeal membrane lipid biosynthesis pathway in Escherichia coli , creating three strains that individually produced archaeal lipids ranging from simple, such as DGGGOH (digeranylgeranylglycerol) and archaeol, to complex, such as GDGT (glycerol dialkyl glycerol tetraether). The physiological responses became more pronounced as the hybrid membrane incorporated more complex archaeal membrane lipids. In particular, biosynthesis of GDGT induced a pronounced SOS response, accompanied by cellular filamentation, explosive cell lysis, and ATP accumulation. Thus, bacteria seem to be able to incorporate simple archaeal membrane lipids, such as DGGGOH and archaeol, without major fitness costs, compatible with the involvement of hybrid membranes at the early stages of cell evolution and in eukaryogenesis. By contrast, the acquisition of more complex, structurally diverse membrane lipids, such as GDGT, appears to be strongly deleterious to bacteria, suggesting that this type of lipid is an archaeal innovation. Impact statement The formation of archaeal–bacterial hybrid membranes represents possible intermediates in early life evolution and eukaryogenesis. In this study, we synthesized archaeal membrane-spanning lipid GDGT (glycerol dialkyl glycerol tetraether) in bacteria Escherichia coli to create such hybrid membranes and examined their physiological properties. Our results suggest that the formation of hybrid membranes containing simple archaeal-type lipids could be a plausible intermediate stage at major evolutionary transitions, whereas more complex, membrane-spanning lipid GDGT would have been deleterious and probably represent a later archaeal innovation, possibly an adaption to thermal habitats. These findings provide new insights and perspectives into the evolution of membranes at the earliest stages of cellular evolution as well as at eukaryogenesis.
研究领域:
微生物学
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分子生物学
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进化生物学
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PubMed
高影响因子
Waste and Biomass Valorization
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2025
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IF 2.8
Zhou Caixue,Dai Shuxia,Yin Li,Zhuo Yu,Zhao Nannan,Li Dehai,Sun Changyan
Purpose In this paper, we use Pinus pumila (Pall.) Pinecone to extract essential oil and improve its utilization. Methods The study used hydrodistillation to extract essential oil from Pinus pumila (Pall.) pinecone. Inhibition analysis of E. coli using extracted essential oils was evaluated to reveal the inhibitory activity by assessing growth curves, conductivity, nucleic acids, proteins, enzyme activities, and protein synthesis. Results The primary constituents, as shown by GC-MS chemical composition study, were α-pinitol, β-pinene and limonene. The results of the bacteriostatic activity indicated that the inhibitory zone (DIZ) had a diameter of 14.93 ± 0.14 mm, the minimum bactericidal concentration (MBC) was 16.0 µL/mL, and the minimum inhibitory concentration (MIC) was 8.0 µL/mL. Sweeping electron microscopy and laser confocal also demonstrated the disruption of cell wall and cellular enzymes, analyzed by CATase and ATPase activities, which showed a decrease in enzyme activity, and SDS-PAGE analysis showed its ability to inhibit protein synthesis, which leads to cellular damage. Conclusion The present study emphasizes the sustainable use of Pinus pumila (Pall.) Pinecone for economic opening of food preservation in addition to inhibition of spoilage bacteria.
研究领域:
植物学
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微生物学
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食品科学
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DOI
高影响因子
FOOD CONTROL
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2016
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IF 3.388
Na Guo,Yu-Ping Zang,Qi Cui,Qing-Yan Gai,Jiao Jiao,Wei Wang,Yuan-Gang Zu,Yu-Jie Fu
Amomum tsaoko is widely distributed in south-west China as a spice. In present study, the antibacterial activity of Amomum tsaoko essential oil (AEO) against foodborne pathogens was evaluated. The antibacterial activity was determined by measuring the zones of inhibition (ZOI), minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and the time-kill assay. Results showed the susceptibility of foodborne bacteria Escherichia coli ( E. coli ) was excellent with the lowest MIC and MBC values of 3.13 and 6.25 mg/mL, respectively. The probable antibacterial mechanism of AEO was the change in permeability and integrity of the cell membranes leading to leakage of nucleic acids and proteins. Detection of the kinetics of E. coli deactivation in situ showed that AEO has good preservative activities in foods. It is necessary to consider that AEO will become a promising antibacterial additive for food preservative.
研究领域:
食品科学
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微生物学
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植物化学
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DOI
高影响因子
JOURNAL OF ALLOYS AND COMPOUNDS
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2019
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IF 4.175
Rui Guo
The metastable α-Ag 2 MoO 4 was usually prepared under high pressure conditions, but it was successfully prepared by epitaxial growth on WO 3 nanosheets at room temperature under normal pressure in this work. The interfacial structure of α-Ag 2 MoO 4 /WO 3 (AW) was characterized by XRD , SEM, TEM, and Raman spectra to explain the process of phase transformation from the β phase to the α phase. The photocatalytic activities were evaluated by the degradation of methylene blue and Escherichia coli . The results indicate that AW-60 exhibits the best photocatalytic activity. The enhancement in photocatalytic performance is mainly due to the photogenerated electron flow between the components from WO 3 to Ag 2 MoO 4 in AW-60. The initial inactivation of E. coli is due to the decomposition of macromolecules with a typical mass range m / z of 2–7 kDa. The corresponding photocatalytic mechanism was proposed and confirmed by the measurements, such as BET, fluorescence spectrum, free radical trapping and the ζ potential of WO 3 and AW-60.
研究领域:
材料科学
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环境科学
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微生物学
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高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
·
2019
·Vol. 103
·期 9
·3715-3725
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IF 3.67
Wang Chengcheng,Zheng Pu,Chen Pengcheng
Raspberry ketone is an important ingredient in the flavor and fragrance industries. Due to its low content in fruits and vegetables, the production of natural raspberry ketone using heterologous synthesis in microbial strains is recently attracting increased attention. In this work, a heterologous pathway to produce raspberry ketone from p -coumaric acid, including 4-coumarate: CoA ligase (4CL), benzalacetone synthase (BAS), and raspberry ketone/zingerone synthase (RZS1) from plants, was successfully assembled in Escherichia coli . When the RZS1 gene was introduced into E. coli and co-expressed with two other genes, the intermediate 4-hydroxybenzylidene acetone in the pathway was almost completely transformed into a raspberry ketone. Substituting TB medium for M9 medium increased raspberry ketone titers by 3–4 times. Furthermore, the heterologous pathway was partitioned into two modules; module one produced p -coumaroyl-CoA from p -coumaric acid by 4CL, and module two produced raspberry ketone from coumaroyl-CoA by the action of BAS and RZS1. Optimizing the balanced expression of the two modules, it was shown that moderate expression of module one and high expression of module two was the best combination to enhance raspberry ketone production. The engineered strain CZ-8 reached 90.97 mg/l of raspberry ketone, which was 12 times higher than previously reported. In addition, the preferred approach of the heterologous pathway was related to the heterologous genes from different sources; for example, 4CL from Arabidopsis thaliana seemed to be more suitable for raspberry ketone production than that from Petroselinum crispum . This work paves an alternative way for future economic production of natural raspberry ketone.
研究领域:
生物化学
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合成生物学
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PubMed
高影响因子
JOURNAL OF INDUSTRIAL MICROBIOLOGY & BIOTECHNOLOGY
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2016
·Vol. 43
·期 4
·557-566
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IF 2.745
Cao Weijia,Ma Weichao,Zhang Bowen,Wang Xin,Chen Kequan,Li Yan,Ouyang Pingkai
The development of efficient microbial processes for pinocembrin production has attracted considerable attention. However, pinocembrin biosynthetic efficiency is greatly limited by the low availability of the malonyl-CoA cofactor in Escherichia coli. Fatty acid biosynthesis is the only metabolic process in E. coli that consumes malonyl-CoA; therefore, we overexpressed the fatty acid biosynthetic pathway enzymes β-ketoacyl-ACP synthase III (FabH) and β-ketoacyl-ACP synthase II (FabF) alone and in combination, and investigated the effect on malonyl-CoA. Interestingly, overexpressing FabH, FabF or both enzymes in E. coli BL21 (DE3) decreased fatty acid synthesis and increased cellular malonyl-CoA levels 1.4-, 1.6-, and 1.2-fold, respectively. Furthermore, pinocembrin production was increased 10.6-, 31.8-, and 5.87-fold in recombinant strains overexpressing FabH, FabF and both enzymes, respectively. Overexpression of FabF, therefore, triggered the highest pinocembrin production and malonyl-CoA levels. The addition of cerulenin further increased pinocembrin production in the FabF-overexpressing strain, from 25.8 to 29.9 mg/L. These results demonstrated that overexpressing fatty acid synthases can increase malonyl-CoA availability and improve pinocembrin production in a recombinant E. coli host. This strategy may hold promise for the production of other important natural products in which cellular malonyl-CoA is rate limiting.
研究领域:
合成生物学
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代谢工程
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微生物学
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高影响因子
BIOTECHNOLOGY LETTERS
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2015
·Vol. 38
·期 4
·619-627
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IF 1.591
Zhuang Yibin,Jiang Jingjie,Bi Huiping,Yin Hua,Liu Shaowei,Liu Tao
Objectives To produce rosmarinic acid analogues in the recombinant Escherichia coli BLRA1, harboring a 4-coumarate: CoA ligase from Arabidopsis thaliana (At4CL) and a rosmarinic acid synthase from Coleus blumei (CbRAS). Results Incubation of the recombinant E. coli strain BLRA1 with exogenously supplied phenyllactic acid (PL) and analogues as acceptor substrates, and coumaric acid and analogues as donor substrates led to production of 18 compounds, including 13 unnatural RA analogues. Conclusion This work demonstrates the viability of synthesizing a broad range of rosmarinic acid analogues in E. coli , and sheds new light on the substrate specificity of CbRAS.
研究领域:
生物化学
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合成生物学
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PubMed
高影响因子
危险品化学品经营许可证（带存储）