APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
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2021
·Vol. 193
·期 7
·2225-2244
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IF 2.926
Wan Yin,Fan Haowei,Gao Lin,Li Ruyi,Xie Mingyong,Wu ChouFei,Chen Longyan,Fu Guiming
Tannase from Aspergillus niger NL112 was purified 5.1-fold with a yield of 50.44% via ultrafiltration, DEAE-Sepharose Fast Flow column chromatography, and Sephadex G-100 column chromatography. The molecular weight of the purified tannase was estimated as 45 kDa. The optimum temperature and pH for its activity were 45 °C and 5.0, respectively. The results of circular dichroism, FT-IR (Fourier transform infrared) spectroscopy, and fluorescence spectra indicated that high temperature could lead to the change of tannase secondary and tertiary structures. Tannase had a greater affinity for tannic acid at 40 °C with a K m value of 2.12 mM and the greatest efficiency hydrolysis ( K cat / K m ) at 45 °C. The rate of inactivation ( k ) increased with the increase of temperature and the half-life ( t 1/2 ) gradually decreased. It was found to be 1.0 of the temperature quotient ( Q 10 ) value for tannic acid hydrolysis by tannase. The thermodynamic parameters of the interaction system were calculated at various temperatures. The positive enthalpy ( ΔH ) values and decreasing ΔH values with the increase of temperature indicated that the hydrolysis of tannase was an endothermic process. Our results indicated that elevated temperature could change the tertiary structure of tannase and reduce its thermostability, which caused a gradual decrease of tannase activity with an increase in temperature.
研究领域:
生物化学
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微生物学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2018
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IF 3.909
Ruyi Li,Guiming Fu,Chengmei Liu,David Julian McClements,Yin Wan,Shaoman Wang,Ting Liu
The tannase (from Aspergillus niger ) was immobilised by glutaraldehyde conjugation to amino-functionalised chitosan-coated magnetic nanoparticles (Fe 3 O 4 -CS nanoparticles). Fourier-transform infrared spectroscopy and thermo-gravimetric analysis showed that chitosan was coated on the surface of magnetic nanoparticles. Transmission electron microscopy indicated that the synthesised nanoparticles (Fe 3 O 4 -CS) were almost spherical or ellipsoidal with an average diameter of 5.97 ± 1.25 nm. The stability and functionality of free and immobilised tannase were compared. Both forms of tannase exhibited the same optimal temperature of 30 °C, whereas the optimal pH value of immobilised tannase (pH 4.5) was lower than that of the free tannase (pH 5.5). The pH and thermal stabilities of immobilised tannase were significantly better than those of free tannase. Immobilised tannase retained over 50% of its initial activity after repeated utilisation for eight cycles. Furthermore, the immobilised tannase effectively improve the clarity and colour of black and green tea infusions. These results showed that amino-functionalised Fe 3 O 4 -CS nanoparticles are an efficient carrier for immobilising tannase, and immobilised tannase can be used in the clarification of tea infusion.
研究领域:
生物技术
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食品科学
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DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2011
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IF 2.816
Jian Yao,Xin Jiong Fan,Yi Lu,Yu Huan Liu
A novel gene (designated as tan410) encoding tannase was isolated from a cotton field metagenomic library by functional screening. Sequence analysis revealed that tan410 encoded a protein of 521 amino acids. SDS−PAGE and gel filtration chromatography analysis of purified tannase suggested that Tan410 was a monomeric enzyme with a molecular mass of 55 kDa. The optimum temperature and pH of Tan410 were 30 °C and 6.4. The activity was enhanced by addition of Ca2+, Mg2+ and Cd2+. In addition, Tan410 was stable in the presence of 4 M NaCl. Chlorogenic acid, rosmarinic acid, ethyl ferulate, tannic acid, epicatechin gallate and epigallocathchin gallate were efficiently hydrolyzed by recombinant tannase. All of these excellent properties make Tan410 an interesting enzyme for biotechnological application.
研究领域:
微生物学
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生物化学
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分子生物学
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DOI
PubMed
高影响因子
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
·
2025
·
IF 3.5
Qianyu Shen,Jiaxin You,Zhike Xie,Junhui Zhang,Quancheng Zhou,Zheng Ruan
BACKGROUND Phenolic acids, such as chlorogenic acid (CGA) and rosmarinic acid (RA), are added to plant-based beverages as nutritional supplements to enhance their health benefits. However, these compounds can also interfere with the astringency-reducing effect of tannase. This study employed electronic tongue analysis, enzyme inhibition kinetics, spectroscopy, molecular docking and molecular dynamics simulations to investigate the inhibitory mechanisms of CGA and RA on tannase-mediated deastringency. RESULTS Our research results indicate that CGA can inhibit tannase-mediated deastringency. It, along with RA, inhibits tannase activity in a non-competitive manner and quenches its intrinsic fluorescence through static quenching. The binding of CGA and RA to tannase led to the exposure of aromatic amino acid residues and a more polar microenvironment. Fourier transform infrared spectroscopy showed that CGA and RA reduced the α -helix and β -turn content in tannase, while increasing the unordered coil content. Molecular docking and dynamics simulations revealed that CGA and RA bind tightly to tannase primarily through hydrogen bonds and van der Waals interactions, occupying the substrate-binding site and thus inhibiting tannase's astringency-reducing activity. Additionally, other polyphenols, such as epicatechin, hesperidin and naringin, were also found to inhibit tannase activity. CONCLUSION The study demonstrated that CGA and RA inhibit the astringency-removal activity of tannase, offering important mechanistic insights for the development of plant-based beverages and deastringency techniques. © 2025 Society of Chemical Industry.
研究领域:
食品科学
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生物化学
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分子生物学
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PubMed
高影响因子
MICROSCOPY RESEARCH AND TECHNIQUE
·
2016
·Vol. 79
·期 9
·880-889
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IF 1.13
Guobin Xia,Chunfang Lin,Songbai Liu
A large scale isolation and purification of theaflavin (TF) and epigallocatechin (EGC) has been successfully developed by tannase-mediated biotransformation combining high-speed countercurrent chromatography. After tannase hydrolysis of a commercially available theaflavins extract (TE), the content of TF and EGC in tannase-mediated biotransformation product (TBP) achieved approximately 3 times enrichment. SEM studies revealed smooth tannase biotransformation and the possibility of recovery of the tannase. A single 1.5 hours' HSCCC separation for TF and EGC employing a two-phase solvent system could simultaneously produce 180.8 mg of 97.3% purity TF and 87.5 mg of 97.3% purity EGC. However, a preparative HPLC separation of maximum injection volume containing 120 mg TBP prepared 11.2 mg TF of 94.9% purity and 7.7 mg EGC of 89.9% purity. HSCCC separation demonstrated significant advantages over Prep HPLC in terms of sample loading size, separation time, environmental friendly solvent systems, and the production.
研究领域:
化学工程
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药学
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分析化学
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PubMed
高影响因子
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
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2022
·Vol. 102
·期 10
·4054-4064
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IF 4.125
Xianxian Dong,Yin Wan,Yanru Chen,Xiaojiang Wu,Yulong Zhang,Mengfei Deng,Wenqin Cai,Xiaodan Wu,Guiming Fu
BACKGROUND Tannase is an enzyme produced by microbial fermentation and is widely used in the food industry; however, the molecular mechanism of tannase production by Aspergillus has not yet been studied. This study was conducted to reveal the differences in Aspergillus carbonarius tannase enzymatic characterization, secondary structures and molecular mechanisms after treatment of the strain with atmospheric and room temperature plasma (ARTP). RESULTS The results showed that the specific activity of tannase was improved by ARTP treatment, and it showed higher thermostability and tolerance to metal ions and additives. The enzymatic characterization and molecular docking results indicated that tannase had a higher affinity and catalytic rate with tannic acid as a substrate after ARTP treatment. In addition, the docking results indicated that Aspergillus tannases may catalyze tannic acid by forming two hydrogen-bonding networks with neighboring residues. RNA-seq analysis indicated that changes in steroid biosynthesis, glutathione metabolism, glycerolipid metabolism, oxidative phosphorylation pathway and mitogen-activated protein kinase signaling pathways might be crucial reasons for the high production of tannase. CONCLUSION ARTP enhanced the yield and properties of A. carbonarius tannase by changing the enzyme structure and cell metabolism. This study provides a theoretical basis for elucidating the molecular mechanism underlying high production of Aspergillus tannases. © 2022 Society of Chemical Industry.
研究领域:
微生物学
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生物化学
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基因组学
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酶学
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分子生物学
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高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2024
·Vol. 25
·期 19
·10547
·
IF 4.9
Xiaona Zeng,Jiabei Song,Shengqiu Tang,Xiaoying Dong,Sheng Chen,Jie Kong,Liyi Chen,Yajuan Li,Guanming Shao,Yung-Hou Wong,Qingmei Xie
Tannins, one of the most common anti-nutritional factors in feed, can be effectively degraded by various enzymes secreted byAspergillus tubingensis(A. tubingensis). The cultivation method of fungi significantly impacts gene expression, which influences the production of enzymes and metabolites. In this study, we analyzed the tannin biodegredation efficiency and the transcriptomic responses ofA. tubingensisin liquid and solid cultures with tannin added. The observed morphology ofA. tubingensisresembled typical fungal hyphae of mycelium submerged and grown in liquid cultures, while mainly spore clusters were observed in solid cultures. Furthermore, the tannin biodegredation efficiency and protein secretion ofA. tubingensisin liquid cultures were significantly higher than in solid cultures. Additionally, 54.6% of the 11,248 differentially expressed genes were upregulated in liquid cultures, including AtWU_03490 (encoding ABC multidrug transporter), AtWU_03807 (ribonuclease III), AtWU_10270 (peptidyl-tRNA hydrolase), and AtWU_00075 (arabinogalactan endo-1,4-beta-galactosidase). Functional and gene ontology enrichment analyses indicated upregulation in processes including oxidation reduction, drug metabolism, and monocarboxylic acid metabolism. Overall, this study provides insight into the transcriptomic response to tannin biodegradation byA. tubingensisin different cultures and reveals that liquid cultures induce greater transcriptomic variability compared to solid cultures.
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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基因组学
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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）