PLANT DISEASE
·
2022
·Vol. 106
·期 9
·
IF 4.614
Chunwei Wang,Tiankun Duan,Luxin Shi,Xiqian Zhang,Weixin Fan,Meiqin Wang,Jianming Wang,Lu Ren,Xiaojun Zhao,Yan Wang
To develop an effective and environmentally safe strategy to control postharvest gray mold caused by Botrytis cinerea , Bacillus siamensis strain YJ15 isolated from blueberry was used to test the biocontrol potential. It is interesting to find that the volatile organic compounds (VOCs) emitted from strain YJ15 exhibited significant antifungal activity against Botrytis cinerea as well as 11 other plant-pathogenic fungi, with a percentage of mycelial growth inhibition from 74.96 to 92.81%. Additionally, VOCs from strain YJ15 could reduce significantly the disease incidence and lesion diameter with the increasing of fermentation time, indicating great biocontrol potential for controlling blueberry postharvest gray mold. Furthermore, the VOCs were collected by using headspace solid-phase microextraction fiber, and the composition of VOCs was further revealed by using gas chromatography coupled with quadruple mass spectrometry. In total, 24 kinds of VOCs, including 5 alkanes, 2 aldehydes, 3 ketones, 5 alcohols, 1 alkene, 5 acids and esters, 2 aromatic compounds, and 1 sulfur compound, were emitted at 1, 3, 5, and 7 days after inoculation. Among these VOCs, eight antifungal VOCs could inhibit mycelial growth of B. cinerea . It is important to highlight that, although 1-butanol and 3-methyl-1-butanol were the most abundant compounds, 2-ethylhexanol, 1-heptanol, and 1,3-xylene have proved to be more toxic to B. cinerea than 3-methyl-1-butanol, propanethioic acid, 2,2-dimethyl-, ethyl 2-methylbutyrate, 2-heptanone, and 1-butanol, which provide new, promising biofumigants for the control of postharvest gray mold caused by B. cinerea .
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APPLIED AND ENVIRONMENTAL MICROBIOLOGY
·
2022
·Vol. 88
·期 19
·
IF 5.005
Shuai Liu,Yuwei Gao,Lin Quan,Mei Yang,Yong-Zhong Wang,Changjun Hou
An increase in exoenzyme production can be enhanced by environmental stresses such as graphene oxide (GO) stress, but the link between the two events is still unclear. In this work, the effect of GO as an environmental stress factor on exoenzyme (lignocellulolytic enzyme, amylase, peptidase, and protease) biosynthesis was investigated in Bacillus subtilis Z2, and a plausible mechanism by which cytosolic Ca2+ regulates lignocellulolytic enzyme production in B. subtilis Z2 subjected to GO stress was proposed. The filter paper-hydrolyzing (FPase [representing total cellulase]), carboxymethylcellulase (CMCase [representing endoglucanase]), and β-glucosidase activities and extracellular protein concentration of the wild-type strain under 10 μg/mL GO stress were 1.37-, 1.64-, 1.24-, and 1.16-fold those of the control (without GO stress), respectively. Correspondingly, the transcription levels of lignocellulolytic enzyme genes, cytosolic Ca2+ level, and biomass concentration of B. subtilis were all increased. With lignocellulolytic enzyme from B. subtilis used to hydrolyze alkali-pretreated rice straw, the released reducing sugar concentration reached 265.53 mg/g, and the removal rates of cellulose, hemicellulose, and lignin were 52.4%, 30.1%, and 7.5%, respectively. Furthermore, transcriptome data revealed that intracellular Ca2+ homeostasis played a key role in regulating the levels of gene transcription related to the synthesis of lignocellulolytic enzymes and exoenzymes. Finally, the use of Ca2+ inhibitors (LaCl3 and EDTA) and deletion of spcF (a calmodulin-like protein gene) further demonstrated that the overexpression of those genes was regulated via calcium signaling in B. subtilis subjected to GO stress.IMPORTANCE To effectively convert lignocellulose into fermentable sugars, high lignocellulolytic enzyme loading is needed. Graphene oxide (GO) has been shown to promote exoenzyme (lignocellulolytic enzyme, amylase, peptidase, and protease) production in some microorganisms; however, the regulatory mechanism of the biosynthesis of lignocellulolytic enzymes under GO stress remains unclear. In this work, the lignocellulolytic enzyme production of B. subtilis under GO stress was investigated, and the potential mechanism by which B. subtilis enhanced lignocellulolytic enzyme production through the calcium signaling pathway under GO stress was proposed. This work revealed the role of calcium signaling in the production of enzymes under external environmental stress and provided a direction to facilitate lignocellulolytic enzyme production by B. subtilis.
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JOURNAL OF HAZARDOUS MATERIALS
·
2020
·
IF 9.038
Ning Hou,Qiaoruo Wang,Yang Sun,Xianyue Li,Qiuying Song,Xinxin Jiang,Baoxin Li,Xinyue Zhao,Hailian Zang,Dapeng Li,Chunyan Li
In recent years, special attention has been devoted to biodemulsifiers as a new type of environment-friendly demulsifiers. A novel biodemulsifying oxalate decarboxylase (OxdC) secreted by Bacillus mojavensis XH1 is reported in the present study. A genome-wide comparison showed that strains with high demulsification efficiencies all possess alkane degradation genes. An analysis of the differentially expressed genes and proteins induced by different substrates showed that OxdC secreted by XH1 was an effective demulsifier. Moreover, the demulsification ability was verified by prokaryotic gene expression, knockout and complementation analyses. OxdC from XH1 exhibited a strong demulsification capacity and significantly outperformed the model protein Bacillus subtilis 168 OxdC (Yvrk), which shared a high amino acid similarity but showed limited demulsification ability. Based on a comparison of the structural characteristics, the hydrophobic amino acids on the surface of OxdC were identified as a key factor driving the favorable demulsification activity of XH1. The metabolic pathways of XH1 used liquid paraffin and glucose as substrates, illustrating that hydrocarbons are necessary for biodemulsifier secretion. The present study provides new insight into the application of OxdC as an additional genetic resource in biodemulsification.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2020
·
IF 4.192
Lei Wang,Yongmei Xia,Lingqia Su,Jing Wu
γ-Cyclodextrin (γ-CD), a cyclic oligosaccharide containing eight glucose units linked by α-1,4-glycosidic bonds, can be produced from starch using cyclodextrin glycosyltransferase (CGTase). Unfortunately, this enzymatic process produces mixtures of α-, β-, and γ-CD. In this study, amino acid residues in the subsite −3 (T47 and F91) and the central subsite (Y186) of Bacillus clarkii γ-CGTase were modified to improve the γ-CD production. The cyclization activities and product specificities of mutants T47H and F91W were similar to those of the wild-type. The cyclization activities of mutants F91N and F91L were significantly greater than those of the wild-type but their γ-CD product specificities were lower. Finally, the central subsite mutant Y186W displayed a γ-CD specificity (94.6%) significantly greater than that of the wild-type (77.1%). To maximize the γ-CD yield, the effects of added complexing agents were investigated. Among the cyclic complexing agents tested, low-boiling cyclododecanone was the smallest that precipitated with γ-CD. When cyclododecanone was used with Y186W, the total CD yield reached 72.6%, and 96.6% of the product was γ-CD. These results, which represent the highest γ-CD yield ever reported, may provide a way to improve large-scale γ-CD preparation and expand the uses of γ-CD in the future.
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BIOPROCESS AND BIOSYSTEMS ENGINEERING
·
2020
·Vol. 44
·期 4
·727-735
·
IF 2.419
Wang Hongrao,Han Shuran,Wang Jiayi,Yu Shuyu,Li Xiaoyan,Lu Lei
Laccases are versatile oxidases that are capable of decolorizing various synthetic dyes. Recombinant Bacillus amyloliquefaciens laccase was immobilized as magnetic cross-linked enzyme aggregates (M-CLEAs) for application in dye decolorization. Several parameters influencing the activity recovery were evaluated during the synthesis of M-CLEAs. With ammonium sulfate as precipitant, maximum activity was recovered by cross-linking with 0.16% glutaraldehyde for 1 h. The prepared M-CLEAs exhibited improved activity under alkaline conditions. It remained 74% activity after incubation at 60 °C for 5 h. Enhanced tolerance towards NaCl was also observed for the M-CLEAs, with 68% activity remaining in the presence of 1 M NaCl. The immobilized laccase could rapidly decolorize more than 93% of reactive black 5 and indigo carmine in 1 h, while its catalytic efficiency towards reactive blue 19 was relatively low. After four cycles of consecutive reuse, the M-CLEAs could decolorize 92% of indigo carmine. The easy recovery and reusability of M-CLEAs facilitate the potential application of bacterial laccase in dye decolorization.
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BIOCHEMICAL ENGINEERING JOURNAL
·
2021
·
IF 3.978
Yaru Zhang,Jing Ren,Quan Wang,Shuxiang Wang,Shuna Li,Hongya Li
In the current work, studies were conducted for understanding oxidation characteristics and biodegradation potential of a dye-decolorizing peroxidases from Bacillus amyloliquefaciens (BaDyP) for triphenylmethane dye crystal violet (CV). Recombinant BaDyP from Bacillus amyloliquefaciens MN-13 exhibits a broad substrates profile. It can oxidize ABTS, veratryl alcohol, Mn 2+ , azo dye reactive black 5, anthraquinone dye reactive blue 19, but cannot oxidize guaiacol. The maximum BaDyP activity against ABTS was obtained at 30 ℃ and pH 4.0. When CV was treated by BaDyP for 72 h, the degradation percentage of 63.0 % was observed. UV–vis spectra and HPLC/MS analysis indicated that CV degradation occurred via N -demethylation and cleavage of conjugated chromophore structure. The identified degradation products included stepwise N -demethylated intermediates during the N -demethylation of CV and, 4-( N -methylamino)-4′-( N ’-methylamino) benzophenone and 4-( N -methylamino)-4′-amino benzophenone after cleavage of conjugated chromophore structure. Based on these data, a hypothetic mechanism of CV degradation mediated by BaDyP was proposed in this study. Finally, it is worth to be noted that BaDyP possessed high decolorization efficiency (72.3 % removal ADMI) for actual dye effluent. These results demonstrated that BaDyP might be used as an efficient biocatalyst for the enzymatic pretreatment of dyes in wastewater.
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ANTONIE VAN LEEUWENHOEK INTERNATIONAL JOURNAL OF GENERAL AND MOLECULAR MICROBIOLOGY
·
2020
·Vol. 113
·期 10
·1467-1477
·
IF 2.674
Yin Yi-Rui,Sang Peng,Yang Feng-Li,Li Tao,Yang Run-Feng,Liu Hong-Yan,Luo Zong-Long,Li Wen-Jun,Yang Li-Quan
A β-glucosidase gene ( bsbgl1a ) from Bacillus sp. CGMCC 1.16541 was expressed in Escherichia coli BL21 and subsequently characterized. The amino acid sequence shared 83.64% identity with β-glucosidase (WP_066390903.1) from Fictibacillus phosphorivorans . The recombinant β-glucosidase (BsBgl1A) had a molecular weight of 52.2 kDa and could hydrolyze cellobiose, cellotriose, cellotetrose, p-nitrophenyl-β- d -glucopyranoside ( p NPG), and p-nitrophenyl-β- d -xylopyranoside ( p NPX). Optimal activity for BsBgl1A was recorded at 45 °C with a pH between 5.6 and 7.6, and 100% of its activity was maintained after a 24 h incubation between pH 4 and 9. Kinetic characterization revealed an enzymatic turnover (Kcat) of 616 ± 2 s −1 (with cellobiose) and 3.5 ± 0.1 s −1 (with p-nitrophenyl-β- d -glucopyranoside). Interestingly, the recombinant enzyme showed cupric ion (Cu 2+ ), sodium dodecyl sulfate (SDS) and alcohol tolerance at 10 mM for Cu 2+ and 10% for both SDS and alcohol. Additionally, BsBgl1A had high tolerance for glucose (Ki = 2095 mM), which is an extremely desirable feature for industrial applications. Following the addition of BsBgl1A (0.05 mg/ml) to a commercial cellulase reaction system, glucose yields from sugarcane bagasse increased 100% after 1 day at 45 °C. This work identifies a Cu 2+ , SDS, alcohol, and glucose tolerant GH1 β-glucosidase with potential applications in the hydrolysis of cellulose for the bioenergy industry.
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Microorganisms
·
2025
·Vol. 13
·期 3
·536
·
IF 4.2
Lina Yang,Can Tang,Yan Cui,Jianhua Zhang
Menaquinone-7 (MK-7) is recognized for its important biological activity, andBacillus subtilisis the preferred strain for its fermentative production. However, the limited phenotypic diversity among high-yielding strains complicates the development of rapid screening methods. To address this, we utilized the effect of MK-7 on transmembrane potential to develop a high-throughput screening (HTS) strategy for efficiently identifying strains with improved MK-7 production. Among various membrane potential fluorescent dyes tested, Rhodamine 123 was selected for quantifying intracellular MK-7 levels due to its effective staining and minimal impact on cell growth. By optimizing pretreatment protocols and staining conditions, we established an HTS protocol that combines fluorescence-activated cell sorting with HPLC to identify strains with increased MK-7 production. A linear correlation was observed between mean MK-7 content and average fluorescence intensity (R2= 0.9646). This approach was applied to mutant libraries generated through atmospheric room temperature plasma mutagenesis. After three cycles of mutagenesis and screening, the mutant AR03-27 was identified, showing an 85.65% increase in MK-7 yield compared to the original SJTU2 strain. Resequencing analysis revealed that the top three mutants contained mutations in genes related to membrane transport, suggesting their potential role in enhancing MK-7 yield.
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Biomed Research International
·
2014
·
IF 0
Jianhua Hao,Junzhong Liu,Mi Sun
A bacterial strain C5 that can produce new type of marine esterase was isolated and screened from marine sludge. According to 16S rRNA sequence analysis and physiological and biochemical experiments, the strain was identified as Bacillus subtilis. A single isozyme with a molecular weight of 86 kDa was observed by SDS-PAGE and native-PAGE. On this basis, the mechanism of esterase B1 secreted by strain C5 degrading parathion-methyl was explored, and the effects of temperature and pH on the degradation rate were investigated. From the results, p-nitrophenol was one of the degradation products of B1 degrading parathion-methyl, and the best degradation effect could be achieved at the temperature of 40°C and the neutral pH value.
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BIOTECHNOLOGY LETTERS
·
2017
·Vol. 39
·期 8
·1229-1235
·
IF 1.73
Wang Qianqian,Xu Yixiang,Xu Jiaqi,Wang Xudong,Shen Chen,Zhang Yan,Liu Xiufeng,Yu Boyang,Zhang Jian
Objectives To characterize glycosyltransferases from Bacillus subtilis ATCC 6633 and investigate their substrate specificity towards plant polyphenols. Results Among the cloned and expressed six UDP-glycosyltransferases (BsGT1-6), BsGT-1 showed activity with a wide range of polyphenols: morin, quercetin, alizarin, rehin, curcumin and aloe emodin. The gene of BsGT-1 has an ORF of 1206 bp encoding 402 amino acids. The recombinant enzyme was purified to homogeneity by Ni–NTA affinity chromatograph, and its biochemical characteristics were identified by HPLC–UV/MS, 1 H-NMR and 13 C-NMR. BsGT-1 has an MW of approx. 46 kDa as indicated by SDS-PAGE; its activity was optimal at 40 °C and pH 8.5. The Km value of BsGT-1 towards morin was 110 μM. Conclusions BsGT-1 from B. subtilis was cloned. It had high catalytic capabilities towards polyphenols which would make it feasible for the structural modification of polyphenols.
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Heliyon
·
2024
·
IF 3.4
Jingya Zhang,Yifan Wu,Wei Li,Honglin Xie,Jingyan Li,Yongqiang Miao,Zengqi Yang,Yefei Zhou,Xinglong Wang
The increased rate of antibiotic resistance strongly limits the resolution of Salmonella enterica serovar Typhimurium (S. Typhimurium) infection. Therefore, new strategies to control bacterial infections are urgently needed. Bacillus subtilis (B. subtilis) and its metabolites are desirable antibacterial agents. Here, we aimed to evaluate the antibacterial activity of the novel B. subtilis strain GXYX (No: PRJNA940956) crude lipopeptide against S . Typhimurium. In vitro, GXYX crude lipopeptides affected S . Typhimurium biofilm formation and swimming and attenuated the adhesion and invasion abilities of S . Typhimurium toward BHK-21 cells; in addition, it inhibited the mRNA expression of the filA , filC , csgA , and csgB genes, which are related to the adhesion and invasion ability of S . Typhimurium. In vivo, pretreatment with GXYX crude lipopeptide via intragastric administration improved the survival rate by 30%, which was related to reductions in organ bacterial loads and clinical signs in mice. Intragastric administration of GXYX crude lipopeptide significantly downregulated the mRNA levels of TNF-α, IL-1β, IL-12 and IL-6 in response to S . Typhimurium-induced inflammation compared with intraperitoneal injection. Moreover, it significantly improved the intestinal barrier-related gene (ZO-1, claudin-1, occludin-1) mRNA levels in intestinal tissue damaged by S. Typhimurium infection. In conclusion, GXYX crude lipopeptides were effective at reducing S . Typhimurium colonization, laying a foundation for the further development of novel antibacterial agents.
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Microorganisms
·
2025
·Vol. 13
·期 4
·798
·
IF 4.2
Xiaoyu Cao,Yu Zhang,Qianqian Xu,Hai Yan
Hyperuricemia is a metabolic disorder owing to the underexcretion and/or overproduction of uric acid (UA). Recent studies have shown that probiotics have many potential applications as functional foods to ameliorate hyperuricemia. In this study, we have successfully isolatedBacillus paranthracisYC03 with urate-lowering potential from Jiangshui. The in vitro results indicated that YC03 exhibited strong biodegradation capacities toward UA and its precursors (inosine and guanosine). Meanwhile, the cell-free extracts of YC03 were also found to catalyze and remove inosine and guanosine. To further explore the application potential of this strain in developing functional foods, we evaluated its probiotic properties through in vitro assays and whole genome analysis.B.paranthracisYC03 has excellent abilities, with tolerance to acid and bile salt and good adhesion. In addition, hemolytic assays, along with antibiotic resistance and biogenic amine production tests, have also preliminarily confirmed the safety of using YC03 in food. We have also annotated the key enzyme genes, includingauaG,hpt,rih,punAanddeoD, which are involved in the biodegradation of UA and nucleosides. The results of nucleoside biodegradation product detection will be valuable for exploring the metabolic pathway for biodegrading nucleosides with YC03. These findings provide meaningful insights for the development of functional foods to improve hyperuricemia usingB.paranthracisYC03.
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