FOOD CHEMISTRY
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2023
·
IF 8.8
Hao Luo,Yihong Bao,Ping Zhu
Nutritional and functional soy-based milk gains growing attention globally in food industry. However, its poor sensorial attributes, single flavor, and limited substance variety become critical issues in displaying balanced nutrition and multifunction for health. Herein, a novel probiotic lycopene-soy milk was developed by genome edited Bacillus subtilis harboring lycopene biosynthesis cassette with efficient lycopene production of 25.73 ± 1.57 mg/g DCW. Further investigation displayed desirable pH, reducing sugar, protein, total phenolic content and isoflavone for achieved milk than conventional soy milk , implying it with well-balanced nutritional quality . Notably, achieved milk exhibited stronger antioxidant capacity and higher isoflavone bioavailability for functionality. Moreover, it possessed significantly high scores for taste, appearance, and overall acceptability, suggesting its excellent sensorial attributes. To our delight, it is the first time to fortify soy-milk with probiotic and lycopene by genome edited B. subtilis to explore additive effect on improving nutritional value and functionality for food application.
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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PubMed
高影响因子
Bioresources and Bioprocessing
·
2022
·Vol. 9
·期 1
·1-12
·
IF 4.983
Ye Jing,Li Yunjie,Bai Yuqing,Zhang Ting,Jiang Wei,Shi Ting,Wu Zijian,Zhang Yi-Heng P. Job
To mimic the Escherichia coli T7 protein expression system, we developed a facile T7 promoter-based protein expression system in an industrial microorganism Bacillus subtilis. This system has two parts: a new B. subtilis strain SCK22 and a plasmid pHT7. To construct strain SCK22, the T7 RNA polymerase gene was inserted into the chromosome, and several genes, such as two major protease genes, a spore generation-related gene, and a fermentation foam generation-related gene, were knocked out to facilitate good expression in high-density cell fermentation. The gene of a target protein can be subcloned into plasmid pHT7, where the gene of the target protein was under tight control of the T7 promoter with a ribosome binding site (RBS) sequence of B. subtilis (i.e., AAGGAGG). A few recombinant proteins (i.e., green fluorescent protein, α-glucan phosphorylase, inositol monophosphatase, phosphoglucomutase, and 4-α-glucanotransferase) were expressed with approximately 25–40% expression levels relative to the cellular total proteins estimated by SDS-PAGE by using B. subtilis SCK22/pHT7-derived plasmid. A fed-batch high-cell density fermentation was conducted in a 5-L fermenter, producing up to 4.78 g/L inositol monophosphatase. This expression system has a few advantageous features, such as, wide applicability for recombinant proteins, high protein expression level, easy genetic operation, high transformation efficiency, good genetic stability, and suitability for high-cell density fermentation.
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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基因工程
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PubMed
高影响因子
Metabolites
·
2025
·Vol. 15
·期 1
·45
·
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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DOI
PubMed
高影响因子
Jove-Journal of Visualized Experiments
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2024
·
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
INTERNATIONAL JOURNAL OF FOOD MICROBIOLOGY
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2024
·
IF 5
Yingying Sun,Lele Shao,Yana Liu,Bo Zou,Han Wang,Xingmin Li,Ruitong Dai
Bacillus cereus spores pose a significant concern during food processing due to their high resistance to environmental stress. Ohmic heating (OH) is an emerging and alternative heating technology with potential for inactivating such spores. This study evaluated the inactivation effects and the biological property changes of Bacillus cereus spores during OH treatments. OH effectively inactivated spores in milk, orange juice, broth, rice soup, and buffer solution in less time than oil bath heating (OB). A decrease in NaCl content improved spore inactivation at the same temperature. Spores were more sensitive to acid at 80–85 °C with OH treatment. Furthermore, OH at 10 V/cm and 50 Hz could reduce the spore resistance and inhibit an increase in spore hydrophobicity and spore aggregation. Both heating methods resulted in significant dipicolinic acid (DPA) leakage and damage to the cortex and inner membranes of the spores. However, OH at 10 V/cm and 50 Hz had the lowest DPA leakage and inflicted the least damage to the inner membrane. The damage to the spore's inner membrane was considered the primary reason for inactivation by OB and OH treatments. Still, OH at 10 V/cm and 50 Hz might also block the germination or outgrowth of treated spores or cause damage to the spore core.
研究领域:
食品科学
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微生物学
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生物技术
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DOI
PubMed
高影响因子
Foods
·
2024
·Vol. 13
·期 11
·1777
·
IF 4.7
Xianwen Ma,Dian Zou,Anying Ji,Cong Jiang,Ziyue Zhao,Xiaoqi Ding,Zongchen Han,Pengfei Bao,Kang Chen,Aimin Ma,Xuetuan Wei
In this study, a novel strain for degrading chitin was identified asBacillus paralicheniformisHL37, and the key chitinase CH1 was firstly mined through recombinant expression inBacillus amyloliquefaciensHZ12. Subsequently, the sequence composition and catalytic mechanism of CH1 protein were analyzed. The molecular docking indicated that the triplet of Asp526, Asp528, and Glu530 was a catalytic active center. The enzymatic properties analysis revealed that the optimal reaction temperature and pH was 65 °C and 6.0, respectively. Especially, the chitinase activity showed no significant change below 55 °C and it could maintain over 60% activity after exposure to 85 °C for 30 min. Moreover, the optimal host strain and signal peptide were obtained to enhance the expression of chitinase CH1 significantly. As far as we know, it was the first time finding the highly efficient chitin-degrading enzymes inB. paralicheniformis, and detailed explanations were provided on the catalytic mechanism and enzymatic properties on CH1.
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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PubMed
高影响因子
Catalysts
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2026
·Vol. 16
·期 2
·129
·
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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高影响因子
Foods
·
2026
·Vol. 15
·期 3
·575
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IF 6
Yiwei Dai,Huiru Zhao,Jincui Wei,Yingxi Chen,Xinping Lin,Sufang Zhang,Chaofan Ji
Chitosan oligosaccharides (COSs), obtained through the hydrolysis of chitosan, exhibit remarkable antibacterial properties. In pursuit of COSs with enhanced antibacterial activity, the enzymatic characteristics of the chitosanase fromBacillus velezensisYB1534 (BvChi) were investigated. The purified BvChi displayed optimal activity at pH 6.0 and 50 °C and showed the highest hydrolytic activity using colloidal chitosan as a substrate, with the presence of Mn2+. The COSs produced by enzymatic hydrolysis of BvChi exhibited a minimum degree of polymerization (DP) of 2, and their antimicrobial activities against certain pathogenic bacteria (Escherichia coli,Staphylococcus aureus,Salmonella typhi50071, andAeromonas hydrophila) were evaluated. Among them, the 20 min hydrolysate showed the strongest growth inhibition against all these pathogens, demonstrated by the inhibition zone diameters, and its MIC and MBC values towardA. hydrophilawere 0.625 and 1.25 mg/mL, respectively. Thin-layer chromatography (TLC) analysis showed that the hydrolyzed products after 20 min contains more COSs with DP > 5. These findings highlighted the potential of BvChi as a biocatalyst for producing antimicrobial COSs, applicable in food preservation and biomedical fields.
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高影响因子
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
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2026
·Vol. 42
·期 10
·560
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IF 4.7
Xia Ruhan,Han Pengxiang,Zhao Qiangbo,Yao Wenzhi,Jiang JingJing,Dong Miaoyin,Li Mengfei
Astragalus mongholicus is one of the most commonly traditional Chinese herbal medicines in China and other Asian countries. However, the root rot disease caused by Fusarium oxysporum can severely reduce root yield and quality, resulting in significant large economic losses. Although biological control has attracted widespread attention due to its safety, environmental friendliness and sustainability, the biocontrol efficiency and potential mechanism of Bacillus velezensis against root rot of A. mongholicus have been rarely investigated. In the present study, a rhizosphere bacterium GNB3-7 was isolated from the rhizosphere soil of wild A. mongholicus and possessed the strong broad-spectrum antagonistic activity on multiple pathogenic fungi, which was identified as B. velezensis on the basis of whole-genome sequence analyses. Further genomic analyses revealed that the B. velezensis GNB3-7 has 12 biosynthetic gene clusters encoding antifungal secondary metabolites (e.g., surfactin, bacillibactin, bacilysin). In addition, a total of 36 volatile organic compounds (VOCs) were detected by GC-MS, among which benzothiazole, disulfide-dimethyl and 2-nonanone showed the strongest antifungal activities, with an inhibition rate of 99.41%, 96.87% and 92.69%, respectively. In pot experiments, the treatment rate and prevention efficiency of B. velezensis GNB3-7 against A. mongholicus root rot achieved 44.44% and 72.22%, respectively. Notably, the genes involved in plant induced systemic resistance (ISR, e.g., AmNPR1 , AmCHI2 and AmCOI1 ) were significantly up-regulated in treatment and prevention plants, indicating the activation of ISR in A. mongholicus . These results indicated that B. velezensis GNB3-7 could efficiently control root rot by synergistic action involving the production of metabolites and VOCs, activation of plant ISR, which deepens our understanding on the biocontrol mechanism of B . velezensis against root rot in medicinal plant A. mongholicus .
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高影响因子
Journal of Fungi
·
2026
·Vol. 12
·期 3
·
IF 4.3
Yihua Liao,Jiayi Shen,Tian Yang,Huijuan Peng,Tingwei Qi,Yan Li,Chengcheng Li
Fusariumhead blight (FHB), caused byFusarium graminearum, is a fungal disease that severely affects wheat. The mycotoxins it produces, such as deoxynivalenol (DON), pose serious risks to human and animal health. In this study, a biocontrol strain, LYH8, was isolated from local sources in Jingzhou, Hubei Province. Plate confrontation assays demonstrated that LYH8 effectively inhibited the mycelial growth ofF. graminearum, with an inhibition rate of 43%, and induced morphological abnormalities such as hyphal swelling and shrinkage. Based on16S rRNAandgyrBgene sequencing and phylogenetic analysis, LYH8 was identified asBacillus velezensis. In vivo experiments showed that disease severity in wheat coleoptiles and spikes was significantly reduced by treatment with LYH8 by 75–85%, and the accumulation of DON and its deoxynivalenol-3-glucoside (D3G) in grains was decreased by 20–22%. Further transcriptome analysis revealed that it affects pathogen growth by regulating amino acid biosynthesis, ribosomal biosynthesis, carbon metabolism pathways, and the catalytic activities of related genes. In summary, LYH8 significantly controlled FHB through multiple mechanisms, including inhibiting mycelial growth, reducing infection, and blocking toxin synthesis, demonstrating strong biocontrol potential.
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ACS Synthetic Biology
·
2026
·
IF 4.5
Qian Zhong,Zhen Li,Waibin Duan,Peng Lei,Xiaoqi Xu,Hong Xu,Sha Li,Yibin Qiu
Dynamic metabolic regulation facilitates real-time sensing of pathway states and precise redirection of metabolic flux, thereby enhancing the biosynthesis of diverse value-added compounds. Here, we designed an autoinducible molecular switch in Bacillus amyloliquefaciens by integrating a Pantoea alhagi-derived PaSspB-ssrA* proteolytic system with the DegU-RapG-PhrG quorum-sensing (QS) module. The OFF module employs the novel PaSspB-ssrA* system for targeted degradation of ssrA*-tagged proteins with 86% efficiency, and the ON module utilizes a DegU-activated PispA promoter for cell density-dependent gene expression. To improve hyaluronic acid (HA) production and reduce byproduct accumulation, this system was applied by fusing ssrA* tags into the glycolysis-critical genes pfkA and fruA to attenuate glycolytic flux, together with QS-regulated coexpression of PaSspB and SthHL in engineered Δldh/ΔwbpA/ΔnagB strains, achieving 13.5 ± 0.19 g/L of low-molecular-weight HA (2.89 × 104 Da). The autonomous, inducer-free platform enables cost-effective HA biosynthesis in B. amyloliquefaciens, with broad applicability for synthesizing high-value bioproducts.
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FOOD RESEARCH INTERNATIONAL
·
2025
·
IF 8
Jiaxun Li,Keyuan Qin,Yuting Shang,Zhenjun Zhu,Yangfu Li,Junhui Wu,Moutong Chen,Liang Xue,Qingping Wu,Juan Wang,Yu Ding
Bacillus cereus is a significant cause of foodborne illnesses, particularly through starchy foods like rice and pasta, leading to the vomiting symptom. The primary toxin involved, cereulide, is preformed within contaminated food and exhibits high resistance to heat, making it particularly difficult to control. Since cereulide persists even after standard food processing methods, its prevention and control remain critical for the food industry. Isoleucine (Ile), a branched-chain amino acid (BCAA), has been linked to the regulation of cereulide production, but the exact mechanisms remain unclear. Based on untargeted metabolomics analysis using liquid chromatography-mass spectrometry (LC-MS), this study demonstrated that Ile supplementation markedly downregulated the folate biosynthesis pathway in B. cereus . Supplementation with folic acid partially restored cereulide production that had been suppressed by Ile. In addition, RT-qPCR analysis revealed significant changes in the expression of key enzymes involved in folate metabolism and related pathways following Ile exposure, further supporting the notion that Ile inhibits cereulide synthesis by disruption of folate metabolism. Collectively, these findings suggest a novel mechanism by which Ile regulates cereulide production via folate metabolism, providing new insights for the effective prevention and control of cereulide production in B. cereus .
研究领域:
食品微生物学
·
代谢组学
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毒理学
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分子生物学
·
微生物生理学
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）