INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2025
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IF 8.5
Xiaotong Xu,Xiaoshuang Yan,Xuerong Jin,Juyi Li,Qiaoxia Hu,Dong-Hyun Ahn,Chao Zhong
Colanic acid (CA), an exopolysaccharide synthesized by bacteria, is known for its potential to extend the lifespan of nematodes. However, its high viscosity poses practical application challenges, which can be mitigated by employing colanic acid-degrading enzymes (CAEs). This study presents the expression and characterization of a novel CAE from Escherichia phage phi92, used for the production of CA oligosaccharides (CAOSs). Optimal CAE expression conditions were identified as induction at 20 °C for 24 h with 0.1 mM IPTG. The enzyme showed maximal activity at 55 °C and pH 6.0, with stability in the range of 4–50 °C and pH 4.0–7.0. Structural characterization of CAOSs was performed using GC–MS, LC–MS and NMR spectroscopy, revealing the composition of hexasaccharide and dodecasaccharide units, with CAE cleaving the β-1,4 glycosidic linkage between glucose and fucose. CAOSs were found to mitigate oxidative stress and inflammation induced by H 2 O 2 in RAW 264.7 macrophages, inhibiting NO and MDA production while enhancing CAT activity. Additionally, CAOSs modulated the mRNA expression of pro-inflammatory (IL-1β, COX-2) and anti-inflammatory (IL-10, HO-1) factors. This study deepens the understanding of CAE and facilitates the preparation of CA oligomers for applications in food, cosmetics, and pharmaceuticals.
研究领域:
微生物学
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生物化学
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医药科学
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DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
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2025
·
IF 3.8
Zhi Wang,Jiaxing Zhang,Shengping You,Rongxin Su,Wei Qi
The extensive application of polyethylene terephthalate (PET) has led to severe environmental pollution. Enzymatic degradation provides a sustainable solution for PET recycling, but the limited hydrolytic activity of current PET hydrolases hinders its practical application. Here, we designed an Energy-Guided Accumulate Mutation Strategy (EGAMS) for the rational design of PET enzyme PHL7. By applying multiple rounds of evolution to enhance the degradation activity and thermostability of PHL7, we achieved the excellent PET degradation enzyme FlashPETase (PHL7 E148K/T158P/S184E/H185Y ) with 238.8% PET degradation activity of PHL7 and 82.93 °C melting temperature. FlashPETase completely degrades PET at 72 °C with an enzyme loading of 1.5 mg enzyme g PET −1 , without any substrate pretreatment or additional processing. As for PET substrates with high crystallinity, FlashPETase still performs well. Furthermore, molecular simulations revealed the structural mechanisms of FlashPETase’s enhanced activity and stability. In summary, the EGAMS protocol and the variants discussed in this work advance the fields of PET degradation and enzyme engineering, offering valuable insights for future research.
研究领域:
酶工程
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环境科学
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分子生物学
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DOI
高影响因子
Frontiers in Microbiology
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2022
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IF 6.064
Yang Peizhou,Xiao Wei,Lu Shuhua,Jiang Shuying,Jiang Suwei,Chen Jianchao,Wu Wenjing,Zheng Zhi,Jiang Shaotong
Aflatoxin B1 (AFB1) contaminates rice during harvest or storage and causes a considerable risk to human and animal health. In this study, Trametes versicolor AFB1–degrading enzyme (TV–AFB1D) gene recombinantly expressed in engineered E. coli BL21 (DE3) and Saccharomyces cerevisiae. The TV–AFB1D enzymatic characteristics and AFB1 degradation efficiency in contaminated rice were investigated. Results showed that the size of recombinant TV-AFB1D expressing in E. coli BL21 (DE3) and S. cerevisiae was appropriately 77 KDa. The kinetic equation of TV-AFB1D was y = 0.01671x + 1.80756 (R2 = 0.994, Km = 9.24 mM, and Vmax = 553.23 mM/min). The Kcat and Kcat/Km values of TV-AFB1D were 0.07392 (s−1) and 8 M−1 s−1, respectively. The AFB1 concentration of contaminated rice decreased from 100 μg/ml to 32.6 μg/ml after treatment at 32°C for 5 h under the catabolism of TV-AFB1D. S. cerevisiae engineered strains carrying aldehyde oxidase 1 (AOX1) and Cauliflower mosaic virus 35 S (CaMV 35 S) promoters caused the residual AFB1 contents, respectively, decreased to 3.4 and 2.9 μg/g from the initial AFB1 content of 7.4 μg/g after 24 h of fermentation using AFB1-contaminated rice as substrate. The AFB1 degradation rates of S. cerevisiae engineered strains carrying AOX1 and CaMV promoters were 54 and 61%, respectively. Engineered S. cerevisiae strains integrated with TV-AFB1D expression cassettes were developed to simultaneously degrade AFB1 and produce ethanol using AFB1-contaminated rice as substrate. Thus, TV-AFB1D has significant application potential in the AFB1 decomposition from contaminated agricultural products.
研究领域:
食品科学
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微生物学
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生物化学
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DOI
PubMed
高影响因子
Royal Society Open Science
·
2020
·Vol. 7
·期 9
·
IF 2.646
Siqi Liu,Xiaolin Xu,Yanshun Kang,Yingtian Xiao,Huan Liu
Ligninolytic enzymes, including laccase (Lac), manganese peroxidase (MnP) and lignin peroxidase (LiP), have attracted much attention in the degradation of contaminants. Genes of Lac (1827 bp), MnP (1134 bp) and LiP (1119 bp) were cloned from Aspergillus sp. TS-A, and the recombinant Lac (69 kDa), MnP (45 kDa) and LiP (35 kDa) were secretory expressed in Pichia pastoris GS115, with enzyme activities of 34, 135.12 and 103.13 U l−1, respectively. Dyes of different structures were treated via the recombinant ligninolytic enzymes under the optimal degradation conditions, and the result showed that the decolourization rate of Lac on Congo red (CR) in 5 s was 45.5%. Fourier-transform infrared spectroscopy, gas chromatography–mass spectrometry analysis and toxicity tests further proved that the ligninolytic enzymes could destroy the dyes, both those with one or more azo bonds, and the degradation products were non-toxic. Moreover, the combined ligninolytic enzymes could degrade CR more completely compared with the individual enzyme. Remarkably, besides azo dyes, ligninolytic enzymes could also degrade triphenylmethane and anthracene dyes. This suggests that ligninolytic enzymes from Aspergillus sp. TS-A have the potential for application in the treatment of contaminants.
研究领域:
环境科学
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生物技术
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微生物学
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产品关联
DOI
PubMed
高影响因子
POSTHARVEST BIOLOGY AND TECHNOLOGY
·
2013
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IF 2.454
Jianwen Bu,Yucong Yu,Guzhanuer Aisikaer,Tiejin Ying
Mature green cherry tomato fruit were harvested and treated with ultraviolet-C (UV-C) irradiation at a predetermined dose of 4.2 kJ m −2 and stored at 18 °C for 35 days. The effects of UV-C treatment on firmness, cell wall compositions (pectin and cellulose), cell wall ultra-structure, levels of ethylene production, and activities of cell wall degrading enzymes in the fruit were investigated during storage. Major genes involved in cell wall degradation ( PME 2.1, Cel 1, PG cat and Exp 1) were also checked. Fruit firmness was better maintained in UV-C treated fruit corresponding with higher contents of cellulose and acid-soluble pectin. Transmission electron microscopy indicated that UV-C irradiation retarded cell wall disassembly in the pericarp. Ethylene production was significantly inhibited by UV-C treatment. The UV-C treatment also suppressed the transcriptional expression of PME2.1 , Cel1 , PGcat and Exp1 genes, and inhibited pectin methylesterase (PME; EC 3.1.1.11), polygalacturonase (PG; EC3.2.1.15) and cellulase (EC3.2.1.4) activities during storage. These results suggested that the inhibition of ethylene production, which in turn down-regulated expression of genes encoding cell wall degrading enzymes, might be one of the possible mechanisms of UV-C delaying tomato fruit softening.
研究领域:
植物学
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园艺学
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DOI
高影响因子
FOLIA MICROBIOLOGICA
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2022
·Vol. 67
·期 4
·633-640
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IF 2.629
Gao Huihui,Lu Dan,Xing Mingyan,Xu Qing,Xue Feng
Zearalenone (ZEN) is a toxic secondary metabolite of Fusarium sp. commonly found in wheat, corn, and other crops. In addition to economic losses, ZEN can seriously endanger the health of both humans and livestock, thus presenting an urgent need for ZEN-detoxifying enzymes that function in the extreme heat or pH conditions of industrial fermenters. Here, we identify and characterize the activity of the ZEN-degrading enzyme from Exophiala spinifera , ZHD_LD, which shares 60.15% amino acid identity and a conserved catalytic triad with the well-characterized ZEN-detoxifying protein ZHD101 from Clonostachys rosea . Biochemical activity and stability assays indicated that purified recombinant ZHD_LD exhibited high activity against ZEN with optimal reaction conditions of 50 ℃ and pH 7.0–10.0. Structural modeling of the ZHD_LD active site and comparison with ZHD101 revealed its likely mechanism of ZEN degradation. This research provides an industrially valuable candidate enzyme for ZEN detoxification in food and livestock feed.
研究领域:
微生物学
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食品安全
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生物化学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2023
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IF 6.1
Yun-Feng Ma,Lang-Lang Gong,Meng-Qi Zhang,Xuan-Zheng Liu,Huan Guo,J. Joe Hull,Gui-Jun Long,Hong Wang,Youssef Dewer,Fan Zhang,Ming He,Peng He
Insects have evolved an extremely sensitive olfactory system that is essential for a series of physiological and behavioral activities. Some carboxylesterases (CCEs) comprise a major subfamily of odorant-degrading enzymes (ODEs) playing a crucial role in odorant signal inactivation to maintain the odorant receptor sensitivity. In this study, 93 CCEs were annotated in the genome of the German cockroach Blattella germanica, a serious urban pest. Phylogenetic and digital tissue expression pattern analyses identified two antenna-enriched CCEs, BgerCCE021e3 and BgerCCE021d1, as candidate ODEs. RNA interference (RNAi)-mediated knockdown of BgerCCE021e3 and BgerCCE021d1 resulted in partial anosmia with experimental insects exhibiting reduced attraction to ester volatile resources and slower olfactory responses than controls. Furthermore, enzymatic conversion of geranyl acetate by crude male antennal extracts from BgerCCE021e3 and BgerCCE021d1 RNAi insects was also significantly reduced. Our results provide evidence for CCE function in German cockroach olfaction and provide a basis for further exploring behavioral inhibitors that target olfactory-related CCEs.
研究领域:
昆虫学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
FOOD RESEARCH INTERNATIONAL
·
2024
·
IF 7
Chenyu Song,Wei Xu,Cuie Guang,Ting Xue,Wanmeng Mu
Patulin (PAT) is a mycotoxin commonly found in fruits and vegetables, prompting the need for effective removal and detoxification methods, which have garnered significant research attention in recent years. Among these methods, the utilization of microbial-derived enzymes stands out due to their mild operating conditions, specificity in targeted functional groups, and the production of non-toxic by-products, making it a preferred degradation approach. In this study, a novel PAT-degrading enzyme derived from Cyberlindnera fabianii ( Cyfa -SDR) was identified, demonstrating its highest catalytic activity at pH 7.0 and 80 °C against PAT. This temperature tolerance level represents the highest reported for PAT-degrading enzymes to date. The enzyme was further characterized as a short-chain dehydrogenase through analysis of its amino acid composition, conserved GXXXGXG motif, and dependency on NADPH. Moreover, the study evaluated the efficiency of PAT degradation by Cyfa -SDR at varying substrate and enzyme concentrations, surpassing the performance of other PAT-degrading enzymes, thus highlighting its substantial potential for the biological control of PAT. In conclusion, the enzymatic treatment using the PAT-degrading enzyme Cyfa -SDR presents a viable and promising solution for enhancing the quality and safety of fruit juice.
研究领域:
食品科学
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微生物学
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酶学
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产品关联
DOI
PubMed
高影响因子
SCIENTIA HORTICULTURAE
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2019
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IF 1.961
Yonghong Ge,Junhu Zhang,Canying Li,Wenjun Xue,Shiyu Zhang,Jingyi Lv
The purpose of this study was to evaluate the effects of trisodium phosphate (TSP) (0.5 g L −1 ) treatment on softening of harvested jujube fruit during storage at 20 ± 2 °C with a relative humidity of 50–55 %. The results showed that jujube fruit treated with TSP showed a higher firmness. The activity of pectin methylesteras, polygalacturonase, cellulase and β-glucosidase was also suppressed by TSP treatment. The results demonstrated that the content of water soluble pectin in jujube fruit treated with TSP was lower than in the control (dipped in distilled water) during storage. TSP treated fruits had higher pectin content than control fruits. The results of atomic force microscopy showed that water soluble pectin structure treated by TSP was denser compared with the control. Taken together, these results demonstrated that TSP could delay softening of jujube fruit by regulating cell wall-degrading enzymes activities and controlling the pectin substances.
研究领域:
食品科学
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植物生理学
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产品关联
DOI
高影响因子
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
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2023
·Vol. 39
·期 11
·1-17
·
IF 4.1
Song Xiaohui,Li Tang,Gu Hui,Yin Heng
Aspergillus is a well-studied fungal genus that is widely used in the processing of plant biomass in industries. This study investigated the effects of space exposure on the ability of Aspergillus costaricaensis , a filamentous fungus isolated from rotten orange peel, to degrade pectin. These fungal spores were carried into space by the Long March 5B carrier rocket and exposed to cosmic radiation for 79 h. After the flight, these spores were resuscitated, and then the growing strains were screened with pectin as the sole carbon source, and the pectinase activity was evaluated. A mutant with increased biomass accumulation ability and pectin-degrading activity compared to the ground control strain was obtained. Comparative transcriptome analysis revealed that several CAZymes genes were significantly upregulated in the mutant, especially those related to pectin degradation. Among the 44 pectinases identified from the annotated genome, 42 were up-regulated. The activities of these pectinases are able to synergistically break down the structure of pectin. In addition, the expression of some genes involved in metabolism, sugar transport, and stress response was altered. These results imply that space exposure might serve as a potential mutagenesis breeding technique, offering the opportunity to acquire biomass-degrading microbial strains with potential for industrial application.
研究领域:
微生物学
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分子生物学
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生物技术
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DOI
高影响因子
JOURNAL OF ENVIRONMENTAL MANAGEMENT
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2025
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IF 8.4
Qing Qiu,Xuejian Sun,Han Li,Fenglin Zhang,Dandan Zhou,Kejian Tian,Xinwen Zhang,Hongliang Huo
Polystyrene (PS), being resistant to biodegradation, poses a significant environmental challenge. This study isolated highly effective lignin-degrading microbial consortia from samples collected at six sites rich in lignin-degrading bacteria. After 360 days of enrichment, a stable lignin-degrading microbial consortium, LQX-03, was successfully established. LQX-03 demonstrated notable degradation efficiency not only for lignin (21-day degradation rate of 54.6 %) but also for PS (21-day degradation rate of 13.1 %). Importantly, PS-induced LQX-03 communities overlapped with the original lignin communities in 13 genera, revealing a close relationship between the degrading microbial compositions of the two substrates.Additionally, Pseudomonas putida Q1, isolated from LQX-03, exhibited significant capability in simultaneously degrading lignin and PS, achieving degradation rates of 36.1 % and 4.4 %, respectively. The strain was also able to alter the functional groups of PS, increasing its hydrophilicity. Gene and enzyme expression analyses revealed that key lignin-degrading enzymes, such as laccase (CopA) and DyP peroxidase, were significantly upregulated when PS was the sole substrate. Laccase CopA expression increased by 1.76-fold and 1.41-fold, while DyP expression increased by 1.24-fold. These results indicate that these enzymes likely play a crucial role in PS depolymerization and biodegradation. Further molecular docking analysis confirmed that laccase CopA could bind to PS. In summary, this study provides preliminary insights into the potential links between lignin-degrading and plastic-degrading microorganisms and their enzymes. It suggests that the biodegradation of synthetic plastics may rely on ancient natural lignin-degrading enzymes. These findings offer a new perspective and valuable data for developing efficient plastic biodegradation strategies.
研究领域:
环境科学
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微生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
iScience
·
2025
·Vol. 28
·期 12
·
IF 4.1
Yao Di,Wenfei Luan,Lihua Sun,Jing Qi,Hongmei Xia,Fan Li,Niu Zhai
Poly(ε-caprolactone) (PCL) is a widely used synthetic polymer with significant commercial applications, and its degradation and synthesis have become the focus of considerable research. In this study, we purified a PCL-degrading enzyme, PCLase0801, produced by Pseudomonas sp. DS0801. The enzyme was purified to homogeneity and had a molecular weight of 30.4 kDa. It exhibited optimal activity at 40°C and pH 8.0, hydrolyzing PCL into monomers, dimers, and trimers. The enzyme was identified as a lipase and showed good tolerance to organic solvents. Additionally, PCLase0801 catalyzed the ring-opening polymerization of ε-caprolactone, producing PCL with a molecular weight of 6050 g/mol and favorable structural properties. This work provides new insights into the potential applications of PCL-degrading enzymes in PCL treatment and biosynthesis.
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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）