FOOD RESEARCH INTERNATIONAL
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2024
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IF 7
Luyao Fan,Hongchao Dai,Wenyuan Zhou,Lei Yuan,Jia Yang,Zhenquan Yang,Xin-an Jiao
Bacillus licheniformis , a quick and strong biofilm former, is served as a persistent microbial contamination in the dairy industry. Its biofilm formation process is usually regulated by environmental factors including the divalent cation Ca 2+ . This work aims to investigate how different concentrations of Ca 2+ change biofilm-related phenotypes (bacterial motility, biofilm-forming capacity, biofilm structures, and EPS production) of dairy B . licheniformis strains. The Ca 2+ ions dependent regulation mechanism for B . licheniformis biofilm formation was further investigated by RNA-sequencing analysis. Results revealed that supplementation of Ca 2+ increased B . licheniformis biofilm formation in a dose-dependent way, and enhanced average coverage and thickness of biofilms with complex structures were observed by confocal laser scanning microscopy. Bacterial mobility of B. licheniformis was increased by the supplementation of Ca 2+ except the swarming ability at 20 mM of Ca 2+ . The addition of Ca 2+ decreased the contents of polysaccharides but promoted proteins production in EPS, and the ratio of proteins/polysaccharides content was significantly enhanced with increasing Ca 2+ concentrations. RNA-sequencing results clearly indicated the variation in regulating biofilm formation under different Ca 2+ concentrations, as 939 (671 upregulated and 268 downregulated) and 951 genes (581 upregulated and 370 downregulated) in B. licheniformis BL2-11 were induced by 10 and 20 mM of Ca 2+ , respectively. Differential genes were annotated in various KEGG pathways, including flagellar assembly, two-component system, quorum sensing, ABC transporters, and related carbohydrate and amino acid metabolism pathways. Collectively, the results unravel the significance of Ca 2+ as a biofilm-promoting signal for B. licheniformis in the dairy industry.
研究领域:
微生物学
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食品科学
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PubMed
高影响因子
Biotechnology for Biofuels and Bioproducts
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2024
·Vol. 17
·期 1
·1-16
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IF 3.3
Wei Xiaoyu,Yang Lijie,Chen Zhen,Xia Wenhao,Chen Yongbin,Cao Mingfeng,He Ning
Background The structural diversity of extracellular polymeric substances produced by microorganisms is attracting particular attention. Poly-gamma-glutamic acid (γ-PGA) is a widely studied extracellular polymeric substance from Bacillus species. The function of γ-PGA varies with its molecular weight (Mw). Results Herein, different endogenous promoters in Bacillus licheniformis were selected to regulate the expression levels of pgdS , resulting in the formation of γ-PGA with Mw values ranging from 1.61 × 10 3 to 2.03 × 10 4 kDa. The yields of γ-PGA and exopolysaccharides (EPS) both increased in the pgdS engineered strain with the lowest Mw and viscosity, in which the EPS content was almost tenfold higher than that of the wild-type strain. Subsequently, the compositions of EPS from the pgdS engineered strain also changed. Metabolomics and RT-qPCR further revealed that improving the transportation efficiency of EPS and the regulation of carbon flow of monosaccharide synthesis could affect the EPS yield. Conclusions Here, we present a novel insight that increased pgdS expression led to the degradation of γ-PGA Mw and changes in EPS composition, thereby stimulating EPS and γ-PGA production. The results indicated a close relationship between γ-PGA and EPS in B. licheniformis and provided an effective strategy for the controlled synthesis of extracellular polymeric substances.
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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PubMed
高影响因子
BIORESOURCE TECHNOLOGY
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2023
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IF 11.4
Renlu Liu,Jialiang Zhang,Haiyun Fu,Li Yin,Yongsheng Song,Genhe He
Efficient removal of organic dye pollution from contaminated water is a concern in the absorbent applications. In this study, a green biogenic calcium carbonate (BCC) absorbent was fabricated using Bacillus licheniformis for the removal of methylene blue (MB) from water. This was found to have superior adsorption capacity compared with abiotic calcium carbonate (ACC) and operate within a broad pH range from 3 to 9. MB adsorption on BCC was physical and exothermic. The hydrophobic features, rough nanoporous microstructure, and organic–inorganic mesoporous structure of the BCC may all be responsible for its favorable adsorption mass transfer. The adsorption energy of BCC had a more negative value than that of ACC, indicating a stronger MB interaction with BCC with a lower energy barrier. Hydrogen bonding and electrostatic attraction were involved in the adsorption process. Overall, the findings established a theoretical foundation for the application of BCC in remediation of MB-contaminated water.
研究领域:
环境科学
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生物学
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PubMed
高影响因子
Metabolomics
·
2019
·Vol. 15
·期 3
·1-12
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IF 3.167
Yuan Honglun,Xu Yong,Chen Yaozhong,Zhan Yangyang,Wei Xuetuan,Li Lu,Wang Dong,He Penghui,Li Shengqing,Chen Shouwen
Introduction Acetoin serves as a high value-added platform with a broad range of applications, and can be effectively produced by Bacillus licheniformis . However, its toxicity to the producing strain hinders the higher acetoin production, and current knowledge about the acetoin resistance mechanisms of B. licheniformis is quite limited. Objectives To comprehensively investigate the metabolic changes in B. licheniformis under acetoin stress. Methods We used gas chromatography-mass spectrometry based untargeted metabolomics approach to measure the metabolic profiles of B. licheniformis under 20, 40 and 80 g/L acetoin stress. Transcriptional analysis was conducted to verify the metabolomics results. Results A total of 119 metabolites were identified in our experiment. The metabolic responses of B. licheniformis to acetoin stress were as follows: (i) pentose phosphate pathway and tricarboxylic acid (TCA) cycle were negatively affected by acetoin stress. In turn, glyoxylate cycle was activated to supply malic acid. (ii) Acetoin stress induced the accumulation of serine, valine, leucine and protective osmolytes (glycine and proline). (iii) Acetoin stress induced a higher saturated fatty acid ratio, which indicated a lower fluidity of cell membrane that could inhibit the entry of acetoin into cytoplasm. (iv) Synthesis of phosphatidylserine was enhanced, and phosphatidylethanolamine content was probably increased under acetoin stress. Conclusions This study revealed the metabolic perturbations of B. licheniformis to acetoin stress. In response to acetoin stress, glyoxylate cycle was activated, protective osmolytes were accumulated, saturated fatty acid ratio was elevated and synthesis of phosphatidylserine was enhanced in B. licheniformis .
研究领域:
微生物学
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代谢组学
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生物技术
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PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
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2021
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IF 10.588
Yangjian Cheng,Tingting Zhang,Li Zhang,Zhibin Ke,Libor Kovarik,Hailiang Dong
Cerium is a critical element to modern technologies . Nowadays, its increased applications have led to elevated levels in the environment. Cerium recovery by microorganisms has gained a great deal of attention. Here, our research showed that Bacillus licheniformis could be used to recover Ce 3+ from aqueous solution. The adsorption capacity of cerium on this bacterial strain achieved 38.93 mg/g (dry weight) biomass. Adsorption kinetics followed a pseudo-second-order rate model, and adsorption isotherm was fitted well with the Freundlich model. Scanning electron microscope (SEM) observations coupled with X-ray energy dispersive spectroscopy (EDS) analysis revealed a spatial association of Ce with C, N, O, S, and P. Fourier transform infrared spectroscopy (FT-IR) analysis further suggested that the phosphate and carboxyl groups on the cell surface might be responsible for the adsorption of cerium. Furthermore, X-ray diffraction (XRD) and transmission electron microscopy (TEM) with electron energy loss spectroscopy (EELS) suggested that cerium initially occurred on the bacterial cell surface as Ce(OH) 3 , which was mainly converted to monazite (CePO 4 ) and a small amount of CeO 2 overtime. Hydrothermal treatment was used to accelerate the mineralization process of cerium by B. licheniformis . The hydrothermal treatment is conducted for comparative analysis of mineralization process in extreme geological condition.
研究领域:
微生物学
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环境科学
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材料科学
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2019
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IF 4.784
Zhenxiang Xu,Guoqiang Chen,Le Xue,Haibo Zhang,Jiming Wang,Haiying Xiang,Jinglong Li,Kai Zheng
Extraction polysaccharide from microorganism is a research hotspot. In this work, a new type of water-soluble exopolysaccharides (EPS) was isolated from Bacillus licheniformis . Firstly, response surface methodology (RSM), based on a three-level, three-factor, was used to determine optimum conditions for EPS extraction. And RSM analysis indicated optimum condition was at the temperature of 8 °C for 10.44 h with ethanol at a concentration of 79.22% (v/v), the maximum yield of EPS was 3.07 g/mL. Secondly, EPS were seperated using DEAE-Sepharose Fast Flow column chromatography and acquired two polysaccharide fractions, BL-P1 and BL-P2. BL-P1 had larger molecular weight than BL-P2 from structural analyses, because of higher content of mannose , ribose , glucuronic acid , galactose , arabinose and fructose in BL-P2. Moreover, the characterization of BL-P1 and BL-P2 was investigated with Fourier transform infrared spectroscopy (FT-IR) and nuclear magnetic resonance (NMR) spectroscopy, the results indicated that EPS was mainly composed of→3)-α- d -Gal p -(1→, →3,5)-α- l -Ara f -(1→, →3)-β- d -Glc p -(1→, β- d -Glc p -(1 → and→4)-β- l -Fuc p -(1 → 4)-β- d -Xyl p -(1 → 4)-α- l -Rha p (1 → 3) -β- d -Man p -(4 → residues. In vitro antioxidant activity assay, EPS exhibited potent quenching capacities on hydroxyl and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radicals in a dose-dependent manner. Furthermore, BL-P2 had higher activity than BL-P1 in inhibiting α-amylase and α-glucosidase, which would have potential to be applied in nutraceutical and pharmaceutical industries.
研究领域:
微生物学
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生物化学
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药理学
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高影响因子
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
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2021
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IF 3.575
Yongxin Li,Hua Huang,Xinshuai Zhang
1-Deoxy-D-sorbitol, the 1-deoxy analogue of D-sorbitol, has been detected in human urine as well as in natural herbs and spices. Although there are sporadic reports about 1-deoxy-D-sorbitol dehydrogenase, the complete catabolic pathway of 1-deoxy-D-sorbitol remains unsolved. Informed by the promiscuous activities of fructose-6-phosphate aldolase (FSA) which is involved in the sorbitol (glucitol) utilization ( gut ) operon and guided by the large scale bioinformatics analysis, we predicted and then experimentally verified the gut operon encoded by Bacillus licheniformis ATCC14580 is responsible for the catabolism of both D-sorbitol and 1-deoxy-D-sorbitol by in vitro activity assays of pathway enzymes , in vivo growth phenotypes, and transcriptomic studies. Moreover, the phylogenetic distribution analysis suggests that the D-sorbitol and 1-deoxy-D-sorbitol catabolic gene cluster is mostly conserved in members of Firmicutes phylum.
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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基因组学
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PubMed
高影响因子
MOLECULES
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2025
·Vol. 30
·期 5
·1160
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IF 4.6
Yuan Yuan,Guowei Zhao,Jing Lu,Lei Wang,Yawei Shi,Jian Zhang
The alkaline protease fromBacillus licheniformisstrain 2709 (AprE 2709) is widely used in Chinese industries but faces stability challenges under high-temperature conditions. This study employed molecular modeling and mutagenesis to identify Asn residues at positions 61, 160, and 211 as key sites affecting the stability of AprE 2709. By leveraging the additive and cooperative effects of mutations, the mutant enzyme AprE 2709 (N61G/N160G/N211G) was engineered, exhibiting enhanced thermostability and catalytic activity. The mutant demonstrated a 2.89-fold increase in half-life at 60 °C and a 1.56-fold improvement in catalytic efficiency compared to the wild-type enzyme. Structural analysis revealed that the improved thermostability was due to altered electrostatic interactions and strengthened hydrophobic contacts. Targeting Asn residues prone to deamidation presents a promising strategy for improving protein heat tolerance. These findings not only enhance our understanding of enzyme stability but also lay a foundation for future research aimed at optimizing alkaline proteases for diverse industrial applications, particularly in high-temperature processes.
研究领域:
生物化学
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分子生物学
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工业酶学
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PubMed
高影响因子
Food Bioscience
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2025
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IF 5.9
Yangjincan Liu,Yuqing Huang,Haiming Hu,Huabing Yang,Lei Xiong,Junping Zheng,Tianxiang Zhu,Zhigang Zhang,Hongtao Liu
s Chenpi is a traditional medicinal and edible food in China. Chenpi polysaccharide (CP) is one of the main active components of Chenpi. This study established a fermentation process using Bacillus licheniformis to obtain fermented Chenpi polysaccharides (FCP) with a reduced degree of polymerization. FCP is an acidic polysaccharide exhibiting a molecular weight of 3720 Da. The three predominant monosaccharides in FCP were arabinose (31.75%), galactose (24.77%), and galacturonic acid (20.94%). According to comprehensive NMR studies, its main chain primarily comprised two types of residues: →4)-α-GalA p- (1→ and →4)-α-Gal p , while its side chains started with →2)-α-Fuc p- (1→ or →2)-β-Rha p- (1→ residues, which were further linked to extending residues such as →4)-β-Ara p- (1→ or →5)-α-Ara f- (1→, and ultimately terminated with terminal residues T-β-GalA p , T-β-Glc p , or T-α-Ara f . Compared to commercial pectic polysaccharides (PP), FCP exhibited higher solubility, a looser structure, and lower viscosity. In vitro gut microbiota fermentation demonstrated that FCP substantially enhanced the proliferation of beneficial bacteria, including Paraprevotella and Lachnospiraceae , while suppressing pathogenic bacteria like Shigella . Furthermore, FCP enhanced short-chain fatty acids (SCFAs) production by gut microbiota, particularly acetic and propionic acids. In the single-strain fermentation experiment, FCP promoted the growth of Lactococcus lactis , Pediococcus acidilactici , Parabacteroides distasonis , and Bacteroides thetaiotaomicron . Metabolomic analysis of Bacteroides thetaiotaomicron fermentation broth further revealed that FCP altered the composition of bacterial metabolites, enhancing the biosynthesis of beneficial metabolites, including propionate, 4'-O-methylglucoliquiritigenin, and cyclokievitone hydrate. This study indicated that B. licheniformis fermentation improves the physical and prebiotic properties of CP, providing new insights for the bioprocessing of Chenpi.
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Metabolites
·
2025
·Vol. 15
·期 1
·45
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IF 3.7
Zhe Liu,Fengxu Xiao,Yupeng Zhang,Jiawei Lu,Youran Li,Guiyang Shi
Purpose: This study aimed to utilize genetically engineeredBacillus licheniformisfor the production of ergothioneine (EGT). Given the value of EGT and the application ofBacillus licheniformisin enzyme preparation production, we cloned the key enzymes (EanA and EanB) fromChlorbium limicola. Through gene alignment, new ergothioneine synthase genes (EanAN and EanBN) were identified and then expressed inBacillus licheniformisto construct strains. Additionally, we investigated the factors influencing the yield of EGT and made a comparison withEscherichia coli. Methods: The relevant genes were cloned and transferred intoBacillus licheniformis. Fermentation experiments were conducted under different conditions for yield analysis, and the stability of this bacterium was also evaluated simultaneously. Results: The constructed strains were capable of producing EGT. Specifically, the yield of the EanANBN strain reached (643.8 ± 135) mg/L, and its stability was suitable for continuous production. Conclusions: Genetically engineeredBacillus licheniformisdemonstrates potential in the industrial-scale production of EGT. Compared withEscherichia coli, it has advantages, thus opening up new possibilities for the application and market supply of EGT.
研究领域:
微生物学
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生物技术
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分子生物学
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PubMed
高影响因子
Jove-Journal of Visualized Experiments
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2024
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IF 1.2
Yushu Tang,Ning Hao
Bacillus licheniformis and bacitracin have a huge application market and value in the fields of medicine, chemistry, aquaculture, agricultural, and sideline products. Therefore, the selection of B. licheniformis with high production of bacitracin is of great importance. In this experimental protocol, Bacillus with a high yield of bacitracin was isolated, purified, and identified from the fresh feces of healthy pigs. The inhibitory effect of secondary metabolite bacitracin on Micrococcus luteus was also tested. Thin-layer chromatography and high-performance liquid chromatography were used for the qualitative and quantitative detection of bacitracin. The physiological and biochemical characteristics of B. licheniformis were determined by relevant kits. The phylogenetic relationships of B. licheniformis were determined and constructed using gene sequence detection. This protocol describes and introduces the standard isolation, purification, and identification process of B. licheniformis from animal fresh feces from multiple perspectives, providing a method for the large-scale utilization of B. licheniformis and bacitracin in factories.
研究领域:
微生物学
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药理学
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农业科学
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水产养殖
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生物技术
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PubMed
高影响因子
Catalysts
·
2026
·Vol. 16
·期 2
·129
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IF 4
Jing Yan,Junbing Tao,Fengxu Xiao,Guiyang Shi,Youran Li
This study systematically characterized the L-cysteine biosynthetic pathway inBacillus licheniformisand demonstrated that exogenous serine supplementation significantly upregulated the expression of pathway-associated genes, confirming serine as the primary precursor driving L-cysteine synthesis. Through targeted gene deletions, we generated knockout strains BL2ΔglyA, BL2ΔsdaAA, BL2ΔmetC, BL2Δ2, and BL2Δ3 to minimize precursor diversion and product degradation. Combinatorial overexpression of the feedback-resistant mutantcysEfand the transportereamAyielded an engineered strain achieving 1.075 g/L L-cysteine in shake-flask fermentation with an 18.69% molar conversion yield. These findings highlight the potential ofB. licheniformisas a platform for sulfur metabolic engineering and provide a sustainable fermentation strategy to replace traditional high-pollution hydrolysis-based L-cysteine production. Additionally, this work reveals fundamental differences in sulfur metabolism networks between Gram-positive and Gram-negative bacteria, elucidating microbial metabolic diversity and the cross-regulatory mechanisms linking sulfur, carbon, and nitrogen metabolism.
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