Systems Microbiology and Biomanufacturing
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2023
·
IF 0
Sun Tingting,Liu Di,Zhang Linmeng,Fan Taiping,Cai Yujie
(2R,3R)-BDO is an important bio-based four-carbon platform compound and widely used in chemical, food, pharmaceutical, fuel and aerospace fields . Serratia marcescens has been shown to be an effective strain for producing BDO. However, there have been few reports on the metabolic engineering of this strain to efficiently produce (2R,3R)-BDO using xylose. In this study, the endogenous strong promoter was screened and modified, and combined with the screened RBS. (2R,3R)-BDO was successfully produced by combining the selected promoters and RBS with the overexpressed key enzymes (ALS, ALSD, BDH, GLF) of (2R,3R)-BDO production. The optimum fermentation temperature and pH were 37 ℃ and 6.0, respectively, and the optimized yield reached 6.0 g/L. After batch fermentation, the (2R,3R)-BDO yield reached 36.6 g/L. This provided a good idea for efficient production of (2R,3R)-BDO from xylose.
研究领域:
生物工程
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微生物学
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生物化学
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DOI
Frontiers in Bioengineering and Biotechnology
·
2019
·
IF 5.122
Pan Xuewei,Sun Changhao,Tang Mi,Liu Chao,Zhang Jianing,You Jiajia,Osire Tolbert,Sun Yang,Zhao Youxi,Xu Meijuan,Yang Taowei,Rao Zhiming
Serratia marcescens, a gram-negative bacterium, found in a wide range of ecological niches can produce several high-value products, including prodigiosin, althiomycin, and serratamolide. Among them, prodigiosin has attracted attention due to its immunosuppressive, antimicrobial, and anticancer properties. However, the regulatory mechanisms behind prodigiosin synthesis in Serratia marcescens remains limited. Here, a transposon mutant library was constructed to identify the genes related to prodigiosin synthesis, and BVG90_02415 gene encoding a peptidoglycan synthesizing enzyme D-Ala-D-Ala carboxypeptidase DacA was found to negatively regulates prodigiosin synthesis. Quantitative measurements revealed that disruption of dacA increased prodigiosin production 1.46-fold that of the wild-type strain JNB5-1 in fermentation medium. By comparing differences in cell growth, pigA gene expression level, cell morphology, membrane permeability, and intracellular prodigiosin concentration between wild-type strain JNB5-1 and dacA mutant SK4-72, results revealed that the mechanism for hyper-producing of prodigiosin by the dacA mutant was probably that dacA disruption enhanced prodigiosin leakage, which in turn alleviated feedback inhibition of prodigiosin and increased expression of pig gene cluster. Collectively, this work provides a novel insight into regulatory mechanisms of prodigiosin synthesis and uncovers new roles of DacA protein in regulating cell growth, cell morphology, and membrane permeability in Serratia marcescens. Finally, this study offers a new strategy for improving production of high-value compounds in Serratia marcescens.
研究领域:
微生物学
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生物化学
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分子生物学
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DOI
PubMed
高影响因子
ACS Synthetic Biology
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2025
·
IF 3.9
Di Liu,Long Wang,Linbo Gou,Yao Lu,Yongai Ma,Songsong Yao,Tai-Ping Fan,Huaxiang Deng,Yujie Cai
Terpenoids are valuable chemicals used across industries. Methanol, a nonsugar-based feedstock, offers an eco-friendly approach to terpenoid production. In this study, a hybrid methylotrophic pathway was engineered in Serratia marcescens (S. marcescens) HBQA7, which is tolerant to both methanol and terpenoid compounds. The pathway utilizes bacterial methanol dehydrogenase (Mdh) and yeast dihydroxyacetone synthase (Das) to produce monoterpenes and sesquiterpenes from methanol and xylose. 13C labeling experiments identified the optimal enzyme pair: Mdh from Acinetobacter gerneri and Das from Pichia angusta, achieving 7.63% 13C enrichment of the central metabolic intermediate pyruvate. Deletion of pentose phosphate pathway genes (rpiAB) enhanced methanol utilization, achieving 22.99% 13C enrichment. Optimization of the mevalonate (MVA) biosynthetic pathway enabled the production of 5.12 g/L mevalonate in shake flask culture from methanol and xylose. Further construction of a haloarchaea-type MVA pathway enabled the production of geraniol (574.12 mg/L) and (−)-α-bisabolol (1256.41 mg/L) in shake flask cultures. This study demonstrates the first methanol conversion into valuable terpenoids in S. marcescens.
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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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2024
·
IF 5.7
Yao Lu,Di Liu,Long Wang,Yongai Ma,Tai-Ping Fan,Huaxiang Deng,Yujie Cai
(−)-α-Bisabolol exhibits analgesic, anti-inflammatory, and skin-soothing properties and is widely applied in the cosmetic and pharmaceutical industries. The use of plant essential oil distillation or chemical synthesis to produce (−)-α-bisabolol is both inefficient and unsustainable. Currently, the microbial production of (−)-α-bisabolol mainly relies on Escherichia coli and Saccharomyces cerevisiae as chassis strains; however, high concentrations of (−)-α-bisabolol have certain toxicity to the strain. This study uses synthetic biology and metabolic engineering strategies to redesign a solvent-tolerant Serratia marcescens for the efficient production of (−)-α-bisabolol. By introducing the Haloarchaea-type mevalonate (MVA) pathway and the (−)-α-bisabolol biosynthesis pathway, we successfully constructed a strain capable of producing (−)-α-bisabolol. The coexpression of the chaperone protein DnaK/J significantly enhanced the soluble expression of the (−)-α-bisabolol synthase, resulting in a 10% increase in (−)-α-bisabolol titer. Furthermore, knockout of the PhoA gene, which reduced the formation of the byproduct farnesol (FOH), further increased the (−)-α-bisabolol titer to 3.21 g/L. In a 5 L bioreactor, S. marcescens achieved a final (−)-α-bisabolol titer of 30.2 g/L, representing the highest titer reported to date. This research provides guidance for the production of (−)-α-bisabolol in nonmodel microorganisms without the requirement for induction.
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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产品关联
DOI
PubMed
高影响因子
BIOFOULING
·
2021
·
IF 3.209
Huai-Zhi Luo,Jin-Wei Zhou,Bing Sun,Huan Jiang,Shi Tang,Ai-Qun Jia
Serratia marcescens NJ01, a Gram-negative bacterium, can infect tomato leaves and cause chlorosis and wilting. The present study evaluated the quorum sensing (QS) and biofilm inhibitory effects of seven carboline compounds against S. marcescens NJ01 at 20 μg ml−1, and subsequently focused the study on norharmane as this had the best inhibitory activity. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis confirmed the down-regulation of QS and biofilm related genes bsmA, bsmB, fimA, fimC, flhD, pigA, pigC and shlA on exposure to norharmane. Fourier-Transform Infrared Spectroscopy (FT-IR) analysis showed a reduction in the major components of the exopolysaccharide (EPS) matrix such as nucleic acids, proteins and fatty acids, which are involved in forming the tertiary structure of biofilms. Norharmane exposure also enhanced the susceptibility of the biofilm to ofloxacin. Hence, norharmane has the potential for use as an antibiotic adjuvant to enhance the efficacy of conventional antibiotics to reduce pathogenic bacterial infections.
研究领域:
微生物学
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药理学
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植物病理学
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产品关联
DOI
PubMed
高影响因子
Nature Communications
·
2025
·Vol. 16
·期 1
·1-16
·
IF 15.7
Gao Zhou,Rensing Christopher,Wang Jie,Shen Chenhui,Elzaki Mohammed Esmail Abdalla,Li Xiaoyun,Tan Jinfang,Jiang Xiaoqian
RNA interference is an eco-friendly alternative to chemical pesticides, yet its efficacy in lepidopterans like Spodoptera frugiperda ( S. frugiperda ) is limited by poor uptake. Here, we report on ZIF-8 polydopamine nanoparticles that protect dsRNA against enzymatic degradation and active the endocytic/phagosome pathways for increased uptake. Furthermore, the uptake of nano-enabled dsRNA induces the overgrowth of Serratia marcescens , this reduces the S. frugiperda reactive oxygen species (ROS) immune response, increasing the effects of plant’s natural defenses, further inhibiting Enteroccous mundtii growth. This work shows the synergistic potential of nanoparticles for influencing the gut bacteria to prevent resistance mechanisms and for RNAi delivery for pest management.
研究领域:
昆虫学
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分子生物学
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农业科学
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微生物学
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产品关联
DOI
PubMed
高影响因子
PROCESS BIOCHEMISTRY
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2021
·
IF 3.757
Haiqing Chen,Fan Yu,Nan Shi,Pingping Du,Shuang Liu,Xianzhou Zhang,Jianxin Tan
A novel lipase gene ( lipA ) from Serratia marcescens L1 was cloned and expressed in Escherichia coli . LipA was shown to consist of 614 amino acid residues with seven residues that are different from the other nine reported lipases from S. marcescens . The activity of recombinant lipA based on a 50-mL culture scale approached 28 U/mL, which was more than that of the endogenous extracellular lipase from strain L1. The purified lipA fusion with a His-tag exhibited optimum activity at 30 °C and pH 8.0. Four effective mutants of lipase were obtained by site-directed mutation (SDM). The replacement of residues Arg552 and Gly2 in the enzyme molecule may play an important role in the catalysis characteristics and be responsible for the changes in thermostability and catalytic efficiency in the mutants. Mut G2P had the highest catalytic efficiency ( k cat / K m ) which was enhanced by 2.4-fold compared with its wild-type predecessor. These results indicated that lipA from S. marcescens could be expressed at a high level in E. coli and could be optimized by SDM, which provides a feasible basis for further study of its application.
研究领域:
微生物学
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生物化学
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基因工程
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DOI
高影响因子
Synthetic and Systems Biotechnology
·
2025
·
IF 4.4
Yue Chen,Linbo Gou,Di Liu,Shengfang Wu,Xiuwen Zhou,Tai-Ping Fan,Long Wang,Yujie Cai
(S)-(+)-linalool, a valuable acyclic monoterpenol secondary metabolite of plants, finds extensive applications in the food, flavor and fragrance, pharmaceutical, and daily chemical industries. Microbial synthesis offers two pathways for its production, among which the mevalonate (MVA) pathway derived from halophilic archaea is more widely employed. However, the traditional Haloarchaea-type MVA pathway relies on bifunctional enzyme catalysis, consuming 3 molecules of ATP. Moreover, the catalytic activity of natural linalool synthase (LIS) is generally low, failing to meet industrial requirements. Additionally, linalool exhibits significant toxicity to microbial hosts, thereby limiting the production capacity of conventional chassis microorganisms. To address these bottlenecks, this study implemented systematic optimizations: Firstly, the Archaeal mevalonate pathway was reconstructed by replacing the original bifunctional enzyme with two monofunctional enzymes, successfully reducing ATP consumption to 2 molecules. Secondly, through directed screening and rational design, a high-activity linalool synthase mutant, CsMLIS I331V/I444L , derived from Coriandrum sativum , was obtained. Furthermore, an enzyme fusion strategy was adopted, involving the introduction of a long flexible linker between key genes, which significantly enhanced catalytic efficiency. Finally, S . marcescens HBQA7Δ sIaAB-pyc , a strain previously screened in our laboratory with broad-spectrum tolerance to terpenoids, was selected as the novel chassis cell. Collectively, these efforts resulted in the construction of a microbial cell factory for the efficient synthesis of (S)-(+)-linalool, laying a solid foundation for industrial-scale production.
研究领域:
代谢工程
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合成生物学
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微生物生物技术
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酶工程
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工业生物技术
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产品关联
DOI
高影响因子
Microbiology Spectrum
·
2022
·
IF 9.043
Lujun Yin,Ping-Ping Zhang,Wei Wang,Shi Tang,Shi-Ming Deng,Ai-Qun Jia
Serratia marcescens (S. marcescens) is an environmental bacterium that causes infections with high morbidity and mortality. Notably, infections caused by multidrug-resistant S. marcescens have become a global public health issue. Therefore, the discovery of promising compounds to reduce the virulence of pathogens and restore antibiotic activity against multidrug-resistant bacteria is critical. Quorum sensing (QS) regulates virulence factors and biofilm formation of microorganisms to increase their pathogenicity and is, therefore, an important factor in the formation of multidrug resistance. In this study, we found that 3-phenylpropan-1-amine (3-PPA) inhibited S. marcescens NJ01 biofilm formation and virulence factors, including prodigiosin, protease, lipase, hemolysin, and swimming. The combination of 3-PPA (50.0 μg/mL) and ofloxacin (0.2 μg/mL) enhanced S. marcescens NJ01 sensitivity to ofloxacin. Based on crystalline violet staining, scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM), 3-PPA (50.0 μg/mL) reduced S. marcescens NJ01 biofilm formation by 48%. Quantitative real-time PCR (qRT-PCR) showed that 3-PPA regulated the expression of virulence- and biofilm-related genes fimA, fimC, bsmB, pigP, flhC, flhD, and sodB. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) indicated that 3-PPA affected intracellular metabolites of S. marcescens NJ01, leading to reduce metabolic activity. These results suggested that 3-PPA inhibits the pathogenicity of S. marcescens NJ01 by occluding QS. Thus, 3-PPA is feasible as an ofloxacin adjuvant to overcome multidrug-resistant S. marcescens and improve the treatment of intractable infections.IMPORTANCE Multidrug-resistant bacteria have become a major threat to global public health, leading to increased morbidity, mortality, and health care costs. Bacterial virulence factors and biofilms, which are regulated by quorum sensing (QS), are the primary causes of multidrug resistance. In this study, 3-PPA reduced virulence factors and eliminated biofilm formation by inhibiting QS in S. marcescens NJ01 bacteria, without affecting bacterial growth, thus restoring sensitivity to ofloxacin. Thus, the discovery of compounds that can restore antibiotic activity against bacteria is a promising strategy to mitigate multidrug resistance in pathogens.
研究领域:
微生物学
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药理学
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传染病学
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抗菌素耐药性
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产品关联
DOI
PubMed
高影响因子
FEMS MICROBIOLOGY LETTERS
·
2018
·Vol. 365
·期 24
·
IF 1.735
Wang Youping,Liu Cheng,Wan Juan,Sun Xiaowen,Ma Wei,Ni Hong
An organophosphorus-degrading bacterium MEW06, which exhibited excellent biodegradation capabilities towards 50 mg/L of methyl parathion (MP), paraoxon and dimethoate, was isolated from Sand Lake (Wuhan, China) and identified as Serratia marcescens subsp. marcescens based on physiological–biochemical characteristics and a 16S rDNA sequence-based phylogenetic tree. MEW06 genome contains a 31.09-kDa methyl parathion hydrolase (MPH) (MPHGM004539) that was 54.9% similar to Pseudomonas sp. WBC-3’s MPH. RT–qPCR revealed that mphGM004539 gene expression was significant up-regulated when co-cultured with MP. mphGM004539 without signal peptide (mphGM004539Δsp) was successful cloned and expressed in Escherichia coli BL21 (DE3). Optimized specific enzyme activity of MPHGM004539ΔSP was 5.26 U/mg under 35°C and pH 11.0 conditions when MP as the substrate. Additionally, Co2+, Cd2+and Fe2+ increased the enzyme activity level. MP could be degraded by MPHGM004539ΔSP into p-nitrophenol probably by hydrolyzing the P–O ester bond. Virulence of MP towards Drosophila melanogaster W1118 was reduced by MEW06 or MPHGM004539ΔSP biodegradation. This is the first cloning and characterization of MPH from the organophosphorus-degrading bacterium S. marcescens. MEW06 and its MPH have potential roles in the bioremediation of organophosphorus pesticide-contaminated eco-systems.
研究领域:
微生物学
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环境科学
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生物化学
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毒理学
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DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
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2022
·
IF 6.056
Jin-Wei Zhou,Peng-Cheng Ji,Chang-Yao Wang,Yu-Jie Yang,Xin-Yu Zhao,Hao-Zhe Tang,Shi-Rong Tang
Serratia marcescens NJ01 and Erwinia carotovora , two pathogenic bacteria involved in vegetable spoilage, can persist in leafy vegetables and subsequently form biofilms. The aim of this study was to evaluate the effect of propyl gallate (PG) on the susceptibility of antibiotics against the biofilms of S. marcescens NJ01 and E. carotovora and uncover its anti-virulence mechanism. PG notably inhibited the production of AHLs and enhanced the susceptibility of antibiotics against the biofilm formation of S. marcescens NJ01 and E. carotovora and significantly improved the disrupted effect on the preformed biofilms of S. marcescens NJ01. PG exposure decreased the minimum inhibitory concentrations of these five antibiotics and reduced the production of virulence factors. Further investigation indicated that PG exposure resulted in the downregulation of genes involved in virulence, biofilm formation, and antioxidant enzymes of S. marcescens NJ01. The suppression of antioxidant enzymes caused the enhancement of oxidative stress, increased membrane permeability, and ultimately promoted the susceptibility of bacterial/biofilm cells to antibiotics. The tomato and lettuce infection assay indicated that PG could notably attenuate the pathogenicity of S. marcescens NJ01 and E. carotovora . These findings suggest that PG has the potential to function as an antibiotic accelerant to defend against spoilage pathogens.
研究领域:
微生物学
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食品科学
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植物病理学
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药学
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产品关联
DOI
高影响因子
WATER AIR AND SOIL POLLUTION
·
2026
·Vol. 237
·期 9
·553
·
IF 3.5
Wang Bingyuan,Wang Fengwei,Lu Hongsheng
Microbial degradation has attracted more and more attention to be an effective way to effectively treat pesticide in the environment and save treatment costs. However, the specific degradation mechanism and the influence of microorganisms on the environment of some pesticides are still unclear. In this study, the Serratia marcescens BCD1 (PV133529) which isolated from the blueberry soil showed a 20 days degradation rate of 93.14% and 91.09% for 50 mg/L and 250 mg/L λ-cyhalothrin, respectively. Product analysis revealed the pathway of λ-cyhalothrin degradation by Serratia marcescens BCD1, including the removal of cyano groups and the cleavage of ester bonds. The morphological changes of BCD1 in the presence of different concentrations of λ-cyhalothrin were observed by SEM electron microscopy, and the cell wall of BCD1 was wrinkled and deformed, and the cell length was shortened under the stress of λ-cyhalothrin. This study indicates that BCD1 as a promising microorganism for λ-cyhalothrin residue and improve blueberry quality in a treatment setting. Graphical
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高影响因子
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