Journal of Fungi
·
2026
·Vol. 12
·期 7
·492
·
IF 4.3
Wei Jian,Yuanyuan Li,Yaqian Zhu,Qing Yao,Youcheng Qin,Haiying Liu,Jing Zhang,Guoyang Qiu,Qihang Gui,Zhengwu Zhao
Black spot disease caused byCeratocystis fimbriata(C. fimbriata) is a severe postharvest disease of sweet potatoes. This study evaluated the biocontrol potential ofBacillus velezensis(B. velezensis) L33a against this pathogen. Confrontation assays showed that L33a inhibited mycelial growth by 82.83%. FDA/PI staining and scanning electron microscopy revealed that L33a disrupted cell membrane integrity and caused severe mycelial deformation. Co-culture experiments indicated that L33a altered the expression of key pathogenic genes inC. fimbriata. Volatile organic compounds (VOCs) from L33a inhibited the pathogen by 77.78%, outperforming cell-free supernatant (CFS). VOCs primarily suppressed spore germination, with phenylethanol (PEA) and octanoic acid achieving 100% inhibition. In planta tests on sweet potato tubers showed that both L33a culture and VOCs significantly reduced lesion expansion. Using qPCR analysis, we found that L33a activated defense-related genes in tissues around wounds, particularly those involved in the jasmonic acid (JA) signaling pathway. In summary,B. velezensisL33a effectively controls sweet potato black rot through multiple mechanisms: direct antifungal activity, inhibition of spore germination, modulation of pathogen gene expression, and induction of host defense responses. It represents a promising natural inhibitor for postharvest disease management.
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Microorganisms
·
2025
·Vol. 13
·期 7
·1540
·
IF 4.2
Wenwen Peng,Haocheng Xu,Meiting Zhang,Baoyang Xu,Bing Dai,Caimei Yang
With 80% of bacterial infections occurring as biofilms, biofilm-related infections have evolved into a critical public health concern. Probiotics such asBacillus licheniformishave emerged as promising alternatives, offering new avenues for effective treatment. This study aimed to evaluate the activity oflicheniformisagainst the growth, biofilm formation, motility, and quorum sensing (QS) ofSalmonella typhimurium. Several experiments were conducted: The minimum inhibitory concentration (MIC) ofBacillus licheniformisagainstSalmonella typhimuriumwas determined to be 0.5 mg/mL using the broth microdilution method. The inhibition zone of 100 mg/mL ofB. licheniformisagainstSalmonella typhimuriumwas 19.98 ± 1.38 mm; the time-growth curve showed thatB. licheniformiscan effectively inhibit the growth ofSalmonella typhimurium. In biofilm experiments, at the MIC ofB. licheniformis, the inhibition rate of immature biofilm ofSalmonella typhimuriumwas 86.9%, and it significantly reduced the production of biofilm components (EPS, e-DNA, and extracellular proteases) (p< 0.05). The disruption rate of mature biofilm byB. licheniformisat the MIC was 66.89%, and it significantly decreased the levels of biofilm components (EPS and e-DNA) (p< 0.5). Microscopic observation showed that both the MIC and 1/2 MIC ofB. licheniformiscould reduce the number of bacteria in theSalmonella typhimuriumbiofilm, which was not conducive to the formation and maintenance of the biofilm structure. Swimming/Swarming assays and QS experiments confirmed thatB. licheniformisinhibits the motility ofSalmonella typhimuriumand the secretion of AI-1-type quorum sensing molecules and downregulates the AI-2 quorum sensing system by upregulatinglsrgene expression. These findings suggest thatB. licheniformiscould be a potential antimicrobial agent and biofilm inhibitor.
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CHEMICAL ENGINEERING SCIENCE
·
2025
·
IF 4.3
Chen Shi,Ya-Kun Wang,Long Yan,Yu-Hong Kang,Guang-Hui Liu,Yong Gao,Ya-Nan Zhu,Xian-Yong Wei,Zhang-Feng Zhou
The biodegradation of coal is an effective way of clean and efficient utilization of coal. However, how to improve the degradation rate has been the focus of attention. The effect of combined surfactants Tr-Rh (Tr-octyl phenoxy poly ethoxy, Rh-rhamnolipid) on the degradation of Baolixile coal by Bacillus sp. XK1 was studied. The degradation experiment showed that compared with non-surfactant, the degradation rate of Tr-Rh increased by 25.4 % and the degradation time was shortened by 2 days. The bacterial growth characteristics experiment found that in the presence of Tr-Rh, the highest OD 600 of Bacillus sp. XK1 reached 1.311 and the alkaline protease activity was 93.09 U/mL, indicating that Tr-Rh promoted the growth of Bacillus sp. XK1 , secreting more alkaline proteases, and increasing the contact with coal. The surface property experiments of coal showed that, the contact angle and adsorption rate of coal sample were 16.2 ° and 68.3 % in the presence of Tr-Rh respectively, which indicated that Tr-Rh improved the hydrophilicity and charge of coal surface, and increased the adsorption of bacteria on the coal surface. Furthermore, the degradation products were characterized by GC–MS and FTIR. Compared with non-surfactant, the chain alkanes of the liquid bioproducts increased from 48.22 % to 66.10 % in the presence of Tr-Rh. FITR analysis indicated that Bacillus sp. XK1 , in the presence of Tr-Rh, degraded certain aromatic rings and side-chain groups in coal samples. This study provides a new method and theoretical basis for the efficient biodegradation of coal by microorganisms to produce chain alkanes.
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WATER SCIENCE AND TECHNOLOGY
·
2017
·Vol. 76
·期 1
·147-154
·
IF 1.197
Zhou Wen,Guan Zheng-Bing,Chen Yu,Zhang Feng,Cai Yu-Jie,Xu Cheng-Wen,Chen Xu-Sheng,Liao Xiang-Ru
Given that spore laccase from the Bacillus genus is heat- and alkali-resistant, it is more suitable for industrial applications than fungal laccase. To determine the optimal culture conditions for spore laccase production, the effects of Cu 2+ concentration, oxygen content, and culture time on spore laccase production from Bacillus pumilus W3 were investigated. The optimal production parameters were 0.2 mM of Cu 2+ , 200 rpm shaking speed, 100 mL liquid loading, and 5 days of cultivation. Spore laccase was efficiently immobilized on amino-functionalized celite. When used in dye decolorization, the immobilized spore laccase removed 84.15% of methyl green and 69.70% of acid red 1 after 48 h of treatment. Moreover, the immobilized spore laccase retained 87.04% of its initial decolorization activity after six cycles in the decolorization of acid red 1. These insights into the culture conditions and immobilization of spore laccases should be useful in the development of spore laccase as a biocatalyst in the treatment of textile wastewater.
研究领域:
生物催化
·
环境生物技术
·
酶工程
·
工业微生物学
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ACS Synthetic Biology
·
2026
·
IF 4.5
Yunlong Wu,Yuhao Sun,Huan Miao,Xiaole Xia,Ling Gao
The biosynthesis of flavin adenine dinucleotide (FAD), an essential redox coenzyme, is limited by inefficient substrate utilization and low catalytic activity. To address this, an “Uptake-Catalysis” system was constructed in Bacillus subtilis for FAD synthesis from riboflavin (RF). First, the Streptomyces davawensis transporter (RibM) was coexpressed with native FAD synthase (BsFADs) at a 2:1 ratio, which increased RF consumption to 51%. Subsequently, the isothermal compressibility perturbation engineering (ICPE) strategy was adopted, generating the optimum mutant BsFADsP61W+R202Y, which increased catalytic efficiency (kcat/Km) toward RF and FMN by 83% and 73%, respectively. Mechanistic analysis revealed that this improvement was due to a 39% shortening of RF substrate channel. Finally, FAD titer peaked at 563 mg/L with a conversion rate (cr-FAD) of 92% in shaking-flask fermentation optimization. This work achieved a 225-fold increase in FAD production, validating the effectiveness of the uptake-catalysis synergistic strategy, thereby establishing an efficient and scalable synthesis paradigm.
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Microbial Cell Factories
·
2022
·Vol. 21
·期 1
·1-13
·
IF 6.352
Xin Qinglong,Chen Yudan,Chen Qianlin,Wang Bin,Pan Li
Background Bacillus amyloliquefaciens is generally recognized as food safe (GRAS) microbial host and important enzyme-producing strain in the industry. B.amyloliquefaciens LB1ba02 is a production strain suitable for secreting mesophilic α-amylase in the industry. Nevertheless, due to the low transformation efficiency and restriction-modification system, the development of its CRISPR tool lags far behind other species and strains from the genus Bacillus . This work was undertaken to develop a fast and efficient gene-editing tool in B.amyloliquefaciens LB1ba02. Results In this study, we fused the nuclease-deficient mutant Cas9n (D10A) of Cas9 with activation-induced cytidine deaminase (AID) and developed a fast and efficient base editing system for the first time in B. amyloliquefaciens LB1ba02. The system was verified by inactivating the pyrF gene coding orotidine 5'-phosphate decarboxylase and the mutant could grow normally on M9 medium supplemented with 5-fluoroorotic acid (5-FOA) and uridine (U). Our base editing system has a 6nt editing window consisting of an all-in-one temperature-sensitive plasmid that facilitates multiple rounds of genome engineering in B. amyloliquefaciens LB1ba02. The total editing efficiency of this method reached 100% and it achieved simultaneous editing of three loci with an efficiency of 53.3%. In addition, based on the base editing CRISPR/Cas9n-AID system, we also developed a single plasmid CRISPR/Cas9n system suitable for rapid gene knockout and integration. The knockout efficiency for a single gene reached 93%. Finally, we generated 4 genes ( aprE , nprE , wprA , and bamHIR ) mutant strain, LB1ba02△4. The mutant strain secreted 1.25-fold more α-amylase into the medium than the wild-type strain. Conclusions The CRISPR/Cas9n-AID and CRISPR/Cas9n systems developed in this work proved to be a fast and efficient genetic manipulation tool in a restriction-modification system and poorly transformable strain.
研究领域:
微生物学
·
基因工程
·
生物技术
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Food Bioscience
·
2023
·
IF 5.2
Chen Ji,Yufang Gao,Xuedi Huang,Fujie Yan,Ting Yu
Microencapsulation is a promising and feasible method to help probiotics against adverse situation and overcome storage instability. In this study, we prepared Bacillus smithii XY1 microcapsules by spray drying. Firstly, wall materials of the microcapsules were screened and the ratio between wall materials and protective agent was optimized. The results showed that 9% gum arabic and 5% soluble starch along with 4% skim milk powder were the best condition for spray drying through response surface method . Through scanning electron microscope (SEM), bacteria were successfully encapsuled in the wall materials and microcapsules presented complete structure without cracks. Compared to non-encapsulated bacteria, encapsulation increased tolerance of B. smithii XY1 in gastrointestinal environment, improved shelf time and showed a better effect on alleviating intestine inflammation of zebrafish induced by dextran dulfate sodium (DSS). Thus, this encapsulation approach may have the potential application for improving efficacy of B. smithii XY1 and other probiotics.
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RSC Advances
·
2022
·Vol. 12
·期 53
·34208-34216
·
IF 4.036
Yilan Zhang,Xiaoguang Ying,Bo Liu,Bo Yang,Xiao Li
Nanofiber membranes have recently been considered as promising supports for the immobilization of microorganisms due to the simplicity and cost-effectiveness of electrostatic spinning technology and the ability to control fiber morphology, such as obtaining higher surface area and porosity. In this study, electrospun polyvinyl alcohol/sodium alginate/attapulgite (PVA/SA/ATP) nanofiber membrane was prepared as support for immobilized Bacillus cereus LY-1 for diesel degradation in an aqueous medium and a significant improvement in diesel removal efficiency was realized. The effect of modified ATP concentration on diesel removal was investigated. The results showed that the nanofiber membranes complexed with cetyl trimethyl ammonium bromide (CTAB) and 1% ATP (w/w) had the best capacity for diesel removing. When the initial diesel concentration was 2 g L−1, about 87.8% of diesel was removed by the immobilized LY-1 cells after 72 h. Immobilization of bacteria improves the ability of bacteria to survive in adverse environments. Immobilized LY-1 cells maintain the nature to remove diesel at high salinity or pH range of 6–9. Furthermore, the reusability of the LY-1 cells-immobilized PVA/SA/CTAB–ATP nanofiber membrane was tested. A diesel removal rate of 64.9% could be achieved after 4 times of use. PVA/SA/CTAB–ATP nanofibrous membranes with immobilized LY-1 cells are feasible, economical and environmentally friendly for remediation of diesel contamination in the aqueous medium, and have potential applications in the future.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2023
·
IF 8.2
Shiyi Wang,Yiwen Zhao,Shufen Mao,Jiang Zhu,Yangyang Zhan,Dongbo Cai,Xin Ma,Dong Wang,Shouwen Chen
Disulfide bonds in proteins have strongly influence on the folding efficiency by constraining the conformational space. The inefficient disulfide bond formation of proteins is the main limiting factor of enzyme activity and stability. This study aimed to increase the activity of disulfide-bond-containing proteins via promoting disulfide bonds formation in Bacillus licheniformis . Initially, the glutamate decarboxylase GAD from Escherichia coli was selected as the model protein and introduced into the B. licheniformis . Then, the disulfide isomerase and oxidoreductase from different sources were excavated and overexpressed successively to improve the catalytic efficiency of GAD. The final engineered B. licheniformis showed significantly improved GAD specific activity (from 10.4 U/mg to 80.0 U/mg), which also presented perfect adaptability for other disulfide-bond-containing proteins, for instance, UDP-glucosyltransferase from Arabidopsis thaliana . Taken together, our work demonstrated that the activity of GAD in B. licheniformis was regulated by the disulfide bonds formation status and provided a promising platform for the expression of disulfide-bond-containing proteins.
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ENVIRONMENTAL POLLUTION
·
2023
·
IF 8.9
Guoqiang Zhao,Jing Zhou,Yanning Tian,Qifeng Chen,Dongmei Mao,Jianchun Zhu,Xing Huang
Fomesafen is a diphenyl ether herbicide used to control the growth of broadleaf weeds in bean fields. The persistence, phytotoxicity, and negative impact on crop rotation associated with this herbicide have led to an increasing concern about the buildup of fomesafen residues in agricultural soils . The exigent matter of treatment and remediation of soils contaminated with fomesafen has surfaced. Nevertheless, the degradation pathway of fomesafen in soil remains nebulous. In this study, Bacillus sp. Za was utilized to degrade fomesafen residues in black and yellow brown soils. Fomesafen's degradation rate by strain Za in black soil reached 74.4%, and in yellow brown soil was 69.2% within 30 days. Twelve intermediate metabolites of fomesafen were identified in different soils, with nine metabolites present in black soil and eight found in yellow brown soil. Subsequently, the degradation pathway of fomesafen within these two soils was inferred. The dynamic change process of soil bacterial community structure in the degradation of fomesafen by strain Za was analyzed. The results showed that strain Za potentially facilitate the restoration of bacterial community diversity and richness in soil samples treated with fomesafen, and there were significant differences in species composition at phylum and genus levels between these two soils. However, both soils shared a dominant phylum and genus, Actinobacteriota, Proteoobacteria, Firmicutes and Chloroflexi dominated in two soils, with a high relative abundance of Sphingomonas and Bacillus . Moreover, an intermediate metabolite acetaminophen degrading bacterium, designated as Pseudomonas sp. YXA-1, was isolated from yellow brown soil. When strain YXA-1 was employed in tandem with strain Za to remediate fomesafen contaminated soil, the degradation rate of fomesafen markedly increased. Overall, this study furnishes crucial insights into the degradation pathway of fomesafen in soil, and presents bacterial strain resources potentially beneficial for soil remediation in circumstances of fomesafen contamination.
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2022
·
IF 8.025
Fengshu Li,Xin Hu,Liying Qin,Haoshuai Li,Yan Yang,Xiuli Zhang,Jinren Lu,Yiming Li,Mutai Bao
Although exopolysaccharide (EPS) has been applied to various fields, EPS for UVR-mediated oxidative stress repair still needs further exploration. In this study, a novel EPS was isolated from the fermentation medium of Bacillus sp. QDR3-1 and its yield was 4.8 g/L (pH 8.0, 12 % glucose, 30 °C and 6 % NaCl). The pure fraction (named EPS-M1) was purified by DEAE-cellulose and Sephadex G-100 column. EPS-M1 was a heteropolysaccharide composed of Man, Glc, Gal, and Fuc with a molecular weight of 33.8 kDa. Scanning electron microscopy (SEM) observed a rough surface and reticular structure of EPS-M1, and EPS-M1 formed spherical aggregates in aqueous solution observed in atomic force microscopy (AFM). Thermal analysis revealed that the degradation temperature of EPS-M1 was 306 °C. Moreover, methylation and NMR analysis determined that EPS-M1 was consisted of →3)-Man p -(1→, →2,6)-Man p -(1→, →4,6)-Glc p -(1→, →3)-Glc p -(1→, →4)-Gal p -(1→, →4)-Fuc p -(1→, and T-Man p -(1→. Furthermore, the cytotoxicity and the repair ability of UVR-mediated cell damage of EPS-M1 were studied with L929 cells. The results showed that EPS-M1 had good biocompatibility and it could mitigate UVR-mediated cell damage by regulating the levels of cellular reactive oxygen species (ROS), depolarization of mitochondrial membrane potential (MMP) and Caspase-3/7 activity. Overall, the structure analysis and the protective effects of EPS against L929 cells exposed to UVR provided an experimental basis for EPS in practical applications.
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INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2022
·Vol. 23
·期 17
·9663
·
IF 6.208
Yawen Zhou,Linshu Jiao,Juan Shen,Huibing Chi,Zhaoxin Lu,Huawei Liu,Fengxia Lu,Ping Zhu
Low catalytic activity is a key factor limiting the widespread application of type II L-asparaginase (ASNase) in the food and pharmaceutical industries. In this study, smart libraries were constructed by semi-rational design to improve the catalytic activity of type II ASNase fromBacillus licheniformis. Mutants with greatly enhanced catalytic efficiency were screened by saturation mutations and combinatorial mutations. A quintuple mutant ILRAC was ultimately obtained with specific activity of 841.62 IU/mg andkcat/Kmof 537.15 min−1·mM−1, which were 4.24-fold and 6.32-fold more than those of wild-type ASNase. The highest specific activity andkcat/Kmwere firstly reported in type II ASNase fromBacillus licheniformis. Additionally, enhanced pH stability and superior thermostability were both achieved in mutant ILRAC. Meanwhile, structural alignment and molecular dynamic simulation demonstrated that high structure stability and strong substrate binding were beneficial for the improved thermal stability and enzymatic activity of mutant ILRAC. This is the first time that enzymatic activity of type II ASNase fromBacillus licheniformishas been enhanced by the semi-rational approach, and results provide new insights into enzymatic modification of L-asparaginase for industrial applications.Keywords:L-asparaginase;protein engineering;saturation mutagenesis;catalytic efficiency;molecular dynamics simulation
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危险品化学品经营许可证（带存储）