FOOD CHEMISTRY
·
2017
·
IF 4.529
Lingtian Wu,Shanshan Wu,Juanjuan Qiu,Chuanxue Xu,Sha Li,Hong Xu
A green process and environmentally benign process is highly desirable in the development of enzymatic catalysis. In this work, the shuttle plasmid pHA01 was constructed and the sucrose isomerase (SIase) was expressed in Bacillus subtilis WB800. The optimal nitrogen and carbon sources for SIase expression were yeast extract (15 g/L) and un-pretreated cane molasses (UCM, 20 g/L), respectively. After the UCM fed, the whole cell activity reached 5.2 U/mL in a 7.5 L fermentor. Optimum catalytic temperature and pH of whole cell were 35 °C and 5.5, respectively. Although the biologic membrane reactor (BMR) system consecutively worked for 12 batches, the sucrose conversion remained higher than 90%, indicating the BMR system had a greater operational stability. Furthermore, isomaltulose production using the BMR system with low-cost cane molasses as its substrate not only reduces the production cost and mediates environmental pollution, but also solves the genetic background problem of the non-food-grade strains.
研究领域:
生物技术
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酶工程
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可持续化学
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微生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2022
·Vol. 23
·期 12
·6588
·
IF 6.208
Jiafeng Niu,Ruxue Yan,Juan Shen,Xiaoyu Zhu,Fanqiang Meng,Zhaoxin Lu,Fengxia Lu
Type I L-asparaginase fromBacillus licheniformisZ-1 (BlAase) was efficiently produced and secreted inBacillus subtilisRIK 1285, but its low yield made it unsuitable for industrial use. Thus, a combined method was used in this study to boost BlAase synthesis inB. subtilis. First, fifteen single strong promoters were chosen to replace the original promoter P43, with PyvyD achieving the greatest BlAase activity (436.28 U/mL). Second, dual-promoter systems were built using four promoters (PyvyD, P43, PaprE, and PspoVG) with relatively high BlAase expression levels to boost BlAase output, with the engine of promoter PaprE-PyvyD reaching 502.11 U/mL. The activity of BlAase was also increased (568.59 U/mL) by modifying key portions of the PaprE-PyvyD promoter. Third, when the ribosome binding site (RBS) sequence of promoter PyvyD was replaced, BlAase activity reached 790.1 U/mL, which was 2.27 times greater than the original promoter P43 strain. After 36 h of cultivation, the BlAase expression level in a 10 L fermenter reached 2163.09 U/mL, which was 6.2 times greater than the initial strain using promoter P43. Moreover, the application potential of BlAase on acrylamide migration in potato chips was evaluated. Results showed that 89.50% of acrylamide in fried potato chips could be removed when combined with blanching and BlAase treatment. These findings revealed that combining transcription and translation techniques are effective strategies to boost recombinant protein output, and BlAase can be a great candidate for controlling acrylamide in food processing.Keywords:L-asparaginase;Bacillus subtilis;promoter;dual-promoter systems;ribosome binding site;scale-up fermentation;acrylamide
研究领域:
生物技术
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分子生物学
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食品科学
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微生物学
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DOI
PubMed
高影响因子
FOOD AND BIOPRODUCTS PROCESSING
·
2023
·
IF 4.6
Yulu Du,Yue Wang,Tianyou Cui,Lianpeng Ge,Fan Yu,Mengqian Zhao,Qihang Xu,Mingjie Jin,Bo Chen,Xuwei Long
Biosurfactant surfactin has a high market value and broad application prospects in the food industry and various industrial fields. However, its production efficiency limits its suitability for real applications. In this study, a new Bacillus subtilis (sp.) 50499 strain was isolated from crude oil-contaminated soil and used to efficiently produce surfactin. The initial surfactin titer was 2.25 g/L after three days of fermentation. Adding 2 g/L of L -Leu significantly improved the titer to over 5.5 g/L after optimizing the culture medium at 37 °C. However, adding D -Leu had the opposite effect on surfactin production , and the surfactin titer dropped to below 0.2 g/L. Furthermore, the product of this strain was analyzed to confirm the identity of surfactin. The surfactin obtained in the present study reduced the surface tension of water to 28.6 mN/m at a critical micelle concentration of 20 mg/L. The emulsification index against n -hexane, corn , and crude oil was as high as over 90%. Hence, the present study provides a newly over-producing strain for surfactin production.
研究领域:
微生物学
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生物技术
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DOI
高影响因子
POLYMER INTERNATIONAL
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2020
·Vol. 70
·期 2
·185-192
·
IF 2.574
Yuanyuan Hou,Fei Huang,Hui Yang,Hao Cong,Xi Zhang,Xiaoxu Xie,Haibo Yang,Qiuping Tong,Ning Luo,Ping Zhu,Jianzong Meng
Levansucrase (EC 2.4.1.10) catalyzes both levan polymerization and sucrose hydrolysis. Levan production by recombinant Bacillus subtilis levansucrase SacB-T305A was investigated. The maximum conversion yield (33.09%) of sucrose to levan and levan yield (115.8 g L –1 ) were obtained as 30% (wt/vol) and 40% (wt/vol) initial sucrose, respectively. Under highly acidic conditions (pH 3.5 and below), the polymerization not only shifted distinctly to high molecular weight levan but also enlarged the size of low molecular weight levan. In particular, the proportion of high molecular weight levan to total products increased to 9-fold at pH 3.5. The presence of Fe 2+ , Mg 2+ , Mn 2+ and Zn 2+ ions at common ion strength (5 mmol L –1 ) increased only the size of low molecular weight levan from about 20 kDa to about 35 kDa. Our findings provide a supplementary view about the mechanism of levan molecular weight distribution in enzymatic synthesis and offer a possibility for producing levan of different molecular weights by modulating enzymatic reaction conditions. © 2020 Society of Industrial Chemistry
研究领域:
生物化学
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工业微生物学
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DOI
高影响因子
PROCESS BIOCHEMISTRY
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2021
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IF 3.757
Huabiao Miao,Rui Jiang,Nanyu Han,Yu Ma,Qian Wu,Yuelin Mu,Zunxi Huang
Amylases have been widely applied for starch saccharification in the food, paper, pharmaceutical, and textile industries. In the present study, α-amylase DL-3-4−1 from B. amyloliquefaciens was successfully expressed in B. subtilis. The enzymatic activity exhibited high stability in a pH range from 4.0 to 11.0 and moderate thermostability. Furthermore, excellent stability was observed when incubated with metal ions, detergents, inhibitors, and organic solvents. Resistance against ethanol, urea, and guanidine−HCl was also observed. More importantly, a strategy to enhance the extracellular expression of α-amylase DL-3-4−1 was developed using 173 Sec-type signal peptides (SPs) as a library based on a fast and sequence-independent method. Fourteen SPs were identified with significantly increased yields of DL-3-4−1 via a high-throughput assay. Specifically, the enzymatic activity of the α-amylase-producing strain containing the signal peptide SP5D10 (YomL) was 3.74-fold higher than that of the strain with the wild type signal peptide (87.30 U/OD). In brief, the maximum specific activity of an α-amylase-producing strain containing the YomL signal peptide was 326.45 U/OD. This is the first report of an α-amylase that exhibits ethanol, urea, and guanidine−HCl tolerance. Hence, this α-amylase represents a promising candidate for future use in industry.
研究领域:
生物技术
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酶学
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微生物学
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DOI
高影响因子
Nanomaterials
·
2024
·Vol. 14
·期 4
·340
·
IF 4.4
Xiaozhou Li,Jie Chen,Bin Wu,Zhen Gao,Bingfang He
Exploring an appropriate immobilization approach to enhance catalytic activity and reusability of cellulase is of great importance to reduce the price of enzymes and promote the industrialization of cellulose-derived biochemicals. In this study, Fe3O4magnetic nanoparticles (MNPs) were functionalized with meso-2,3-dimercaptosuccinic acid to introduce carboxyl groups on the surface (DMNPs). Then, melamine–glutaraldehyde dendrimer-like polymers were grafted on DMNPs to increase protein binding sites for the immobilization of processive endoglucanase EG5C-1. Moreover, this dendrimer-like structure was beneficial to protect the conformation of EG5C-1 and facilitate the interaction between substrate and active center. The loading capacity of the functionalized copolymers (MG-DMNPs) for EG5C-1 was about 195 mg/g, where more than 90% of the activity was recovered. Immobilized EG5C-1 exhibited improved thermal stability and increased tolerability over a broad pH range compared with the free one. Additionally, MG-DMNP/EG5C-1 biocomposite maintained approximately 80% of its initial hydrolysis productivity after five cycles of usage using filter paper as the substrate. Our results provided a promising approach for the functionalization of MNPs, enabling the immobilization of cellulases with a high loading capacity and excellent activity recovery.
研究领域:
生物化学
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纳米生物学
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酶工程
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF HAZARDOUS MATERIALS
·
2026
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IF 10.6
Mingliang Zhang,Fanghui Ding,Xi Xiao,Minggen Cheng,Qing Hong,Xin Yan
Enzymatic depolymerization is a potentially efficient and environmentally friendly solution for poly(butylene adipate-co-terephthalate) (PBAT) waste recycling and utilization. However, the depolymerization capacity of reported depolymerases against diverse, real-world commercial PBAT films remains unclear, hindering accurate evaluation of their utility. In this study, a mutant PBAT depolymerase (Tfcut-B3) with enhanced thermal stability and improved activity was obtained through directed evolution of Tfcut-DM. Tfcut-B3 completely depolymerized PBAT white film into terephthalic acid (TPA) within 48 h, yielding 2.0 and 1.3 times more TPA than Tfcut-DM and Tfcut-DM(Q132Y), respectively. Tfcut-B3 completely degraded 8/10 commercial PBAT-based films into TPA within 48 h, with the remaining two (PB-6 and PM-4) being broken down into small fragments; Tfcut-DM was unable to completely degrade any of the films. Tfcut-B3 overexpression and extracellular secretion in Bacillus subtilis was achieved using chromosomal integration; in flask fermentation, a Tfcut-B3 expression level of 274 mg/L was achieved through promoter and host optimization. Tfcut-B3 fermentation supernatant completely degraded up to 64.9 mg of PBAT white film within a 20 mL system at 72 h. These findings provide a theoretical basis for the large-scale application of PBAT depolymerases.
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DOI
高影响因子
FOOD CHEMISTRY
·
2026
·
IF 10.4
Sheng-Dong Chen,Hsin-Ya Tsai,Chen Hsu,Nan-Wei Su
An assay enables high-throughput screening of phosphorylation activity. • Umbelliferone enables activity readout via phosphorylation-induced fluorescence quenching. • The assay was validated with high accuracy, precision, and reproducibility. • Directed evolution using the HTS platform identified a thermostable Bs PPS variant (G145D). To date, enzymatic systems capable of phosphorylating phenolic compounds are rare, and phenolic phosphate synthetase from Bacillus subtilis ( Bs PPS) is distinguished by its ability to regioselectively phosphorylate a broad range of structurally diverse natural polyphenols, a transformation valuable for improving the aqueous solubility and bioavailability of phytochemicals. However, the lack of an efficient high-throughput screening (HTS) platform has limited its engineering and application. Here, we developed a fluorescence-based HTS assay for rapid estimation of Bs PPS activity using umbelliferone as a fluorogenic probe. Phosphorylation of umbelliferone causes pronounced fluorescence quenching, providing an inverse correlation between fluorescence intensity and catalytic activity. The assay was systematically optimized and validated, demonstrating high accuracy, precision, and reproducibility. Application to an error-prone PCR library identified Bs PPS variants with improved thermostability while maintaining catalytic activity. Overall, this assay fills a critical methodological gap and provides a practical foundation for future exploration of related phenolic compound–phosphorylating enzymes. Download: Download high-res image (211KB) Download: Download full-size image
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DOI
高影响因子
Journal of Environmental Sciences
·
2025
·
IF 6.3
Wei Zhang,Xu Zeng,Jiayi Dong,Yibin Su,Erik Jeppesen,Liqing Wang,Sicheng Yin
Microorganism-based biocontrol strategies for harmful cyanobacterial blooms have gained increasing attention due to their ecological compatibility and species-specific targeting. Despite this, research on algicidal bacteria effectiveness against the invasive cyanobacterium Raphidiopsis raciborskii remains limited. Research has been conducted on the Bacillus subtilis strain CH21, which was isolated from Chaohu Lake in China. exhibiting potent indirect anticyanobacterial activity against R. raciborskii . Within 48 h of exposure, the cell-free filtrate of strain CH21 achieved 96.2 % removal efficiency of R. raciborskii cells. Physiological and morphological analyses revealed severe disruption of the photosynthetic apparatus (Fv/Fm and Yield values reduced to zero within 16 h), oxidative stress (elevation in malondialdehyde content and reactive oxygen species, reduction in superoxide dismutase and catalase activity), and cellular damage, including pseudovacuole collapse, the degradation of polyphosphate granules and cytoplasmic leakage. Metabolomic profiling identified surfactin-class lipopeptides as putative bioactive agents, which demonstrated exceptional stability across a wide thermal (25–121 °C) and pH (3–13) range. These findings highlight B. subtilis CH21 as a promising candidate for targeted biocontrol of R. raciborskii blooms, supporting the development of environmentally mitigation technologies to be used in cases where it is difficult to control R. raciborskii by external nutrient loading reduction or if the response to such loading reductions are slow.
研究领域:
微生物学
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环境科学
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应用生态学
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生物技术
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藻类学
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代谢组学
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产品关联
DOI
高影响因子
Microbial Cell Factories
·
2019
·Vol. 18
·期 1
·1-10
·
IF 4.402
Tao Weiyi,Lin Junzhang,Wang Weidong,Huang He,Li Shuang
Background Bioemulsifiers are surface-active compounds, which exhibit advantages including low toxicity, higher biodegradability and biocompatibility over synthetic chemical surfactants. Despite their potential benefits, some obstacles impede the practical applications of bioemulsifiers, including low yields and high purification costs. Here, we aimed to exploit a novel protein bioemulsifier with efficient emulsifying activity and low-production cost, as well as proposed a design-bioemulsifier system that meets different requirements of industrial emulsification in the most economical way. Results The esterase AXE was first reported for its efficient emulsifying activity and had been studied for possible application as a protein bioemulsifier. AXE showed an excellent emulsification effect with different hydrophobic substrates, especially short-chain aliphatic and benzene derivatives, as well as excellent stability under extreme conditions such as high temperature (85 °C) and acidic conditions. AXE also exhibited good stability over a range of NaCl, MgSO 4 , and CaCl 2 concentrations from 0 to 1000 mM, and the emulsifying activity even showed a slight increase at salt concentrations over 500 mM. A design-bioemulsifier system was proposed that uses AXE in combination with a variety of polysaccharides to form efficient bioemulsifier, which enhanced the emulsifying activity and further lowered the concentration of AXE needed in the complex. Conclusions AXE showed a great application potential as a novel bioemulsifier with excellent emulsifying ability. The AXE-based-designer bioemulsifier could be obtained in the most economical way and open broad new fields for low-cost, environmentally friendly bioemulsifiers.
研究领域:
生物化学
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微生物学
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环境科学
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产品关联
DOI
PubMed
高影响因子
BRAZILIAN JOURNAL OF MICROBIOLOGY
·
2022
·Vol. 53
·期 2
·547-556
·
IF 2.214
Sun Xiaodong,Kong Xiangxiang,Li Chunxia,Wang Minjia,Yi Jialin,Deng Zhirui,Niu Bing,Chen Qin
Previous studies have shown that the combination disinfectant, Ortho -phthalaldehyde and benzyldimethyldodecylammonium chloride (ODB), can effectively kill a variety of microorganisms, such as Escherichia coli , Staphylococcus aureus , and Candida albicans . To observe the sporicidal ability and mechanism of ODB for spores, Bacillus subtilis spores were used as the research object in this experiment. TEM images revealed that ODB destroyed the integrity of the coat, cortex, and inner membrane of the spores after 0.5-h treatment, and the nuclear material was also broken and exuded after 4-h treatment. The broken structure led to the release of dipicolinic acid (DPA) in large amount. The results show that B. subtilis spores can be effetely killed by ODB through destroying the structure of the spores.
研究领域:
微生物学
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消毒科学
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生物技术
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DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2023
·
IF 3
Ma Lingling,Wang Ying,Wang Xin,Lü Xin
The process of tobacco aging plays a significant role in enhancing the smoking experience by improving the flavor and quality of tobacco leaves. During natural aging, the metabolic activity of the microbes on the surface of tobacco leaves will be greatly changed. Besides, starch and protein are two of the main macromolecular compounds causing the poor smoking quality of tobacco leaves which to be degraded for better tobacco quality. In this study, a bacterium with the simultaneously degrading ability of starch (degradation rate of 33.87%) and protein (degradation rate of 20%) has been screened out from high-class tobacco leaf and then inoculated into low-class tobacco leaf by solid-state fermentation for quality improvement. The changes in components related to carbon and nitrogen showed that the strain had an obvious effect on the quality improvement of tobacco leaves. After that, GC-MS analyses displayed the volatile flavor compounds which become rich and the flavor has been improved. It has been proved that inoculation solid-state fermentation by dominant strain could improve tobacco quality, as well as instead of the traditional natural aging process which greatly shortens the aging process. The work also offers a helpful strategy for solid-state products for deep fermentation.
研究领域:
生物技术
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食品科学
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农业科学
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）