WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
·
2025
·Vol. 41
·期 4
·1-15
·
IF 4.2
Zhao Jing-yi,Gao Fan,Wu Mengru,Li Yang,Chen Yong,Xiao Zijun
Enantiopure ( S )-2-methylbutanoic acid [( S )-2-MBA] is a high-value chiral compound with applications in fragrances, pharmaceuticals, and agrochemicals. However, conventional chemical synthesis lacks stereoselectivity, while existing biosynthetic methods suffer from low yield and purity. Here, we report a novel microbial process using Bacillus spizizenii ATCC 6633 for efficient ( S )-2-MBA production via L-isoleucine catabolism. Through targeted screening of rhizospheric soil isolates and Bacillaceae strains, ATCC 6633 demonstrated superior performance, producing 3.67 g/L ( S )-2-MBA with 99.32% enantiomeric excess ( ee ) under optimized conditions (45 °C, 8% inoculation, 5 g/L glucose, and 8 g/L L-isoleucine). A 58.92% conversion efficiency was achieved, and a simplified purification process recovered 63.90% product with 97.32% purity. Mechanistic studies suggested glucose depletion triggered ( S )-2-MBA accumulation, aligning with starvation-induced secondary metabolism. This cost-effective, eco-friendly approach eliminates racemic separation steps and harsh reagents, positioning ATCC 6633 as a promising biocatalyst for sustainable ( S )-2-MBA production.
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Food Bioscience
·
2023
·
IF 5.2
Han Hu,Caiyun Wu,Fanglan Ge,Yao Ren,Wei Li,Jiao Li
Soybean meal (SBM), a high-protein animal feed, contains several anti-nutritional factors, which limit its utilization. Poly-γ-glutamic acid (γ-PGA), a biological macromolecule , can significantly improve the health and growth benefits of animal feed. However, there are rare reports about SBM fermentation with γ-PGA-producing bacteria to improve its nutritional quality . In this study, we isolated a glutamic acid-independent poly γ-glutamic acid (γ-PGA) producing Bacillus velezensis SCP024–S-4 bacterial strain and used it for solid fermentation of SBM. The results showed that the maximum γ-PGA yield in fermented soybean meal (FSBM) reached up to 89.64 g/kg without adding glutamic acid . Meanwhile, the content of proteins, free amino acids and small peptides increased by 12.83%, 347.96 μg/g and 108.11 mg/g, respectively. Soy globulin, β-conglutinin and trypsin inhibitor all decreased below 3 mg/g. Mice were fed with CK (control diet), SBM and FSBM respectively. The average daily food intake in the FSBM, SBM, and CK groups were 7.68, 5.25, and 6.36 g, respectively. Compared with the CK group, the mice weight growth rate was significantly higher in the FSBM group. Mice blood and spleen tests showed no negative effect of FSBM. The high-throughput sequencing of mice intestinal microflora indicated that FSBM increased the richness of intestinal microbiota ; especially, the types and quantities of Lactobacillus were significantly increased. This study provides a feasible strategy to improve the nutritional quality of SBM as an animal feed.
研究领域:
微生物学
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动物营养
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发酵技术
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食品科学
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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
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基因工程
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生物技术
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POULTRY SCIENCE
·
2021
·
IF 3.352
Bing Zhang,Haoran Zhang,Yang Yu,Ruiqiang Zhang,Yanping Wu,Min Yue,Caimei Yang
ABSTRACT This study evaluated the effects of Bacillus coagulans ( B. coagulans ) as an alternative to antibiotics on growth performance, antioxidant capacity, immunity function and gut health in broilers. A total of 480 one-day-old broilers were randomly divided into 3 treatments with 8 replicates comprising 20 broilers each. The experiment lasted 42 d. Treatments included: basal diet without antibiotics (NCO); basal diet supplemented with 75 mg/kg chlortetracycline (ANT) ; basal diet supplemented with 5 × 10 9 CFU/kg B. coagulans (BC) . The B. coagulans enhanced body weight ( BW ) and average daily gain compared with the NCO group ( P < 0.05). However, there were no significant differences in average daily feed intake and feed: gain ratio ( F: G ) among three groups ( P > 0.05). The B. coagulans significantly increased catalase, superoxide dismutase, and glutathione peroxidase levels and reduced malondialdehyde levels ( P < 0.05). The serum immunoglobulins (IgA, IgM, and IgY) were significantly higher in the BC group when compared to the NCO and ANT groups ( P < 0.05). The B. coagulans also markedly reduced serum levels of proinflammatory factors (IL-1β, IL-6, and TNF-α) and enhanced anti-inflammatory factor (IL-10) concentrations compared with control group ( P < 0.05). Moreover, compared with the control group, BC significantly inhibited serum xanthine oxidase activity ( P < 0.05). The levels of acetic acid, propionic acid, butyrate, isobutyric acid and valerate in BC group were significantly increased on d 42 compared with the NCO and ANT groups ( P < 0.05). Furthermore, BC significantly altered cecal microbiota by reducing Desulfovibrio and Parasutterella , and by increasing Alistipes and Odoribacter ( P < 0.05, P < 0.05, P < 0.001, P < 0.01, respectively). In conclusion, dietary B. coagulans , when used as an alternative to antibiotics, improved body weight, average daily gain, antioxidant capacity, immunity function and gut health in broilers.
研究领域:
家禽科学
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微生物学
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免疫学
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营养学
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动物科学
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Foods
·
2026
·Vol. 15
·期 4
·753
·
IF 6
Meilin Zhao,Zhihua Huang,Shuoyang Sun,Quan Gan,Shuang Wu,Xiqi Hu,Delei Xu,Pedro Laborda,Bao Tang,Lingtian Wu
Phytophthora capsici , an invasive oomycete pathogen causing blight in crops like cucurbits, tomatoes, and peppers, has led to significant economic losses. Due to the limitations of conventional control methods, research has focused on effective and eco-friendly biocontrol alternatives. The bacterial strain LT-22, identified as Bacillus velezensis , exhibits broad-spectrum antifungal activity and plant growth-promoting potential. This study investigated the biocontrol efficacy of volatile organic compounds (VOCs) from strain LT-22. LT-22 VOCs strongly inhibited P. capsici PC153 growth in vitro and reduced disease symptoms in pepper fruits. Microscopic observations showed that LT-22 VOCs caused significant morphological changes in P. capsici PC153 mycelium by disrupting cell wall and membrane integrity. The treatment increased malondialdehyde (MDA) levels and reduced cellulose content, further indicating that LT-22 VOCs compromise cell integrity to exert antifungal effects. Furthermore, 2-methylbutanoic acid was identified as the primary antifungal VOC, with an EC 50 value of 0.095 µL/mL against P. capsici PC153, lower than the reported plant-derived compounds. Together, these results indicate that strain LT-22 and its VOCs, especially 2-methylbutanoic acid, have significant potential as an effective biocontrol agent for managing Phytophthora blight.
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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
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环境生物技术
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酶工程
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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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JOURNAL OF HAZARDOUS MATERIALS
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2026
·
IF 10.6
Baiyun Jin,Fan Ye,Wei zhou,Guoyu Xiang,Zhoucheng Li,Yu Ding,Wentao Ma,Yuanzhu Mi
Non-covalent interactions predominated by hydrogen bonding are of vital importance to the demulsification of oil-water emulsions. This study reports a Bacillus subtilis -inspired ionic liquid demulsifier (MEB-12) with multiple hydrogen-bonding sites for the treatment of oil-in-water (O/W) emulsions. The two hydrophobic “tails” of this demulsifier facilitate rapid migration within the emulsion and enable precise localization to the oil droplets, while the four symmetric hydrophilic head groups form a multivalent hydrophilic domain. This domain outcompetes interfacial asphaltenes by forming stronger multidentate hydrogen bonds with water molecules, mediated by its abundant ether, ester, and bromide hydrogen-bonding sites, thereby destabilizing the emulsion film. When treating O/W emulsions with an oil content of 0.2% at ambient temperature, MEB-12 at a concentration of 10 mg/L achieved a light transmittance (T %) of 98.4% and an oil removal efficiency (ORE) of 99.95%, demonstrating excellent demulsification performance at low dosage. Meanwhile, MEB-12 exhibited remarkable tolerance toward high salinity. To investigate the competitive adsorption mechanism between demulsifier and asphaltenes, zeta potential, dynamic interfacial tension, droplet coalescence time, and three-phase contact angle were employed as characterization techniques. Additionally, density functional theory was utilized to analyze the electrostatic potential on the molecular surface. Meanwhile, the independent gradient model based on Hirshfeld partition analysis and bond critical point were adopted to explore hydrogen bonding interactions. These characterization and computational simulation results demonstrated that MEB-12 exhibited excellent interfacial activity. Specifically, MEB-12 triggered the reconfiguration of the oil-water interfacial film via non-covalent interactions, thereby disintegrating the rigid film, promoting the coalescence of emulsified droplets, and achieving efficient demulsification.
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Journal of Agriculture and Food Research
·
2026
·
IF 7.2
Zhuoyu Li,Yuchen Li,Nengwu Wang,Guanhong Wu,Xinying Li,Jinqin Deng,Xin Wang,Yan Zhang,Zhaoming Dong
The silkworm ( Bombyx mori ), a well-established model organism in the order Lepidoptera, is widely used to investigate insect immunity. Currently, our understanding of the innate immunity of silkworm is primarily based on genomic and transcriptomic studies. To explore potential antimicrobial proteins/peptides (AMPs), this study infected silkworm pupae with the probiotic Bacillus subtilis and collected hemolymph for proteomic analysis. A total of 669 proteins were identified, of which 214 were significantly changed after infection. GO and KEGG analyses revealed several immune- and metabolism-related processes and pathways, while protein interaction analysis identified clusters associated with immune-related functions as well as clusters associated with protein synthesis. Finally, we mapped the immune pathways of silkworm pupae in response to bacterial infection and found that immune recognition proteins were primarily upregulated in the early stage of infection, whereas immune effector proteins were upregulated in the late stages. This study provides a comprehensive proteomic profile of immune-related proteins in pupal hemolymph and valuable insights for the development and application of AMPs in the silkworm.
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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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3 Biotech
·
2026
·Vol. 16
·期 10
·411
·
IF 3.1
Rahman Mujeeb ur,Shah Junaid Ali,Haroon,Gul Hansa,Nasreen Zahida,Shoaib Muhammad
This study reports the development of a recombinant Bacillus ligniniphilus L1 laccase–rice straw biochar composite and its application in deltamethrin bioremediation. The L1-laccase was heterologously expressed in E. coli BL21 (DE3), purified to a specific activity of 186 U/mg, and covalently immobilized on NaOH- and glutaraldehyde-modified rice straw biochar. Molecular docking against the AlphaFold2-predicted structure (UniProt E9PZ36) yielded a binding affinity of − 6.9 kcal/mol, with one hydrogen bond to MET216 (2.77 Å) and hydrophobic contacts with MET197, MET202, and MET210. However, 100 ns molecular dynamics simulations confirmed a conformationally stable complex (RMSD ~0.42 nm, radius of gyration ~2.21 nm, SASA ~215 nm 2 ). Immobilization shifted the activity optima from pH 7.5 and 45 °C (free) to pH 8.0 and 50 °C (immobilized) and markedly improved stability: the immobilized enzyme retained 74% of its initial activity after 6 days at 4 °C and 18% after 8 catalytic cycles. The apparent Kₘ increased from 0.38 mM (free) to 0.54 mM (immobilized), with kcat values of 178 s⁻ 1 and 130.7 s⁻ 1 , respectively. Under an aggressive deltamethrin load (1000 mg/L), the immobilized laccase achieved 92.3% degradation within 18 h at 50 °C, compared with 68.7% for the free enzyme, producing two major phenolic metabolites (tR = 1.854 and 2.436 min) and one minor unidentified product. These results establish a cost-effective, reusable, and scalable biocatalyst for the bioremediation of pyrethroid-contaminated environments.
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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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