LWT-FOOD SCIENCE AND TECHNOLOGY
·
2024
·
IF 6
Luyu Shan,Jingjing Yang,Yaling Shi,Yalin Ye,Min Zhang,Ying Cui,Huijuan Yu,Yuefei Wang,Xin Chai
Various enzymes secreted by microorganisms during fermentation of Massa Medicata Fermentata (MMF) play a crucial role in conversion of the chemical substances for better clinical application. However, enzyme-driven transformation of chemical compounds during MMF fermentation has not been fully unveiled. Our findings revealed that MMF fermented with a mixture of wheat bran (WB)-flour (2:1) was observed with high enzymatic activities, alongside distinguished appearance. The activities of protease, ester bond hydrolase for cCGA and EF, and glycoside hydrolase all increased with the extended fermentation time of MMF, reaching 14.84×10 −8 ±0.3436×10 −8 , 0.05456×10 −8 ±0.0017×10 −8 , 0.5715×10 −8 ±0.1285×10 −8 , and 15.25×10 −8 ±0.01389×10 −8 Kat/g on the 8th day of fermentation. Moreover, protease has been proven to drive the conversion of WB proteins into amino acids by precolumn phenyl isothiocyanate derivatization. Besides, transformation patterns of ester bond hydrolase on substrate components were revealed, in which cryptochlorogenic acid was decomposed into caffeic acid and quinic acid, and ethyl ferulate was metabolized into ferulic acid. By glycoside hydrolase, amygdalin was transformed into prunasin and benzaldehyde. This study reveals the enzyme-mediated transformation patterns of the focused components during fermentation of MMF, laying foundation for further elucidation of its fermentation mechanism and elevation of quality control.
研究领域:
药理学
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微生物学
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生物化学
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中医药
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DOI
高影响因子
WATER RESEARCH
·
2024
·
IF 11.4
Xin Qi,Jing-Yu Qin,Shaoguo Ru,Jiu-Qiang Xiong
Microalgae-based biotechnology is one of the most promising alternatives to conventional methods for the removal of antibiotic contaminants from diverse water matrices. However, current knowledge regarding the biochemical mechanisms and catabolic enzymes involved in microalgal biodegradation of antibiotics is scant, which limits the development of enhancement strategies to increase their engineering feasibility. In this study, we investigated the removal dynamics of amphenicols (chloramphenicol, thiamphenicol, and florfenicol), which are widely used in aquaculture, by Chlamydomonas reinhardtii under different growth modes (autotrophy, heterotrophy, and mixotrophy). We found C. reinhardtii removed >92 % chloramphenicol (CLP) in mixotrophic conditions. Intriguingly, gamma-glutamyl hydrolase (GGH) in C. reinhardtii was most significantly upregulated according to the comparative proteomics, and we demonstrated that GGH can directly bind to CLP at the Pro77 site to induce acetylation of the hydroxyl group at C3 position, which generated CLP 3-acetate. This identified role of microalgal GGH is mechanistically distinct from that of animal counterparts. Our results provide a valuable enzyme toolbox for biocatalysis and reveal a new enzymatic function of microalgal GGH. As proof of concept, we also analyzed the occurrence of these three amphenicols and their degradation intermediate worldwide, which showed a frequent distribution of the investigated chemicals at a global scale. This study describes a novel catalytic enzyme to improve the engineering feasibility of microalgae-based biotechnologies. It also raises issues regarding the different microalgal enzymatic transformations of emerging contaminants because these enzymes might function differently from their counterparts in animals.
研究领域:
环境生物技术
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生物化学
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微生物学
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产品关联
DOI
PubMed
高影响因子
APPLIED CATALYSIS B-ENVIRONMENTAL
·
2023
·
IF 22.1
Zhixuan Yu,Jinxing Chen,Daiyong Chao,Xiaoxuan Sun,Ling Liu,Shaojun Dong
Conversing proteinaceous enzyme behavior into nanomaterials is intriguing academically, but the threshold from mimicry to accessing sophisticated biological entities has not yet been crossed. Herein, we disclosed the pan-glycoside hydrolases-like (p-GHs-like) activities of Cu 3 P and Cu 2 O nanoparticles and decently confirmed their consistent catalytic mechanism as natural glycoside hydrolase, that is, the precise bond breaking, anomeric carbon configuration and catalytic attack mode. We then introduced p-GHs-like NPs into Escherichia coli to establish a feedback regulatory model for the β- D -glucuronidase (expressed by uidA) that was initially expressed in trace amounts. An upregulation of uidA from 72 to 1000 counts was induced by the p-GHs-like activities via pre-released β-glucuronides derivatives, further contributing to 1–2 orders enhancement of the enzyme production. We unearthed the specific catalytic mechanism to unlock a black box of enzyme-like inorganic feedback on the natural enzyme synthesis process, pushing mimicry leaped to physiological behavior.
研究领域:
纳米技术
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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
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Ting Wu,Huashan Sun,Wenyao Wang,Bin Xie,Zhengjie Wang,Jianqi Lu,Anming Xu,Weiliang Dong,Jie Zhou,Min Jiang
Enzymatic PET recycling has emerged as a promising green solution in addition to mechanical recycling, but low soluble expression levels of the inherently hydrophobic PET hydrolases hinder large-scale applications. Here, we propose a novel strategy for enhanced production of FastPETase in Escherichia coli using co-expression of molecular chaperones from Ideonella sakaiensis . Co-expression of cognate DnaK and DnaJ chaperones significantly increased soluble FastPETase expression (up to 2.5-fold), surpassing commercial chaperone plasmids. Furthermore, a combinatorial approach employing co-expression of DnaK/DnaJ chaperones and fusion of FastPETase with the VNp6-tag significantly boosted FastPETase secretion, yielding over 2 g/L of target protein in a 5-l bioreactor. Notably, the crude FastPETase in fermentation broth displayed comparable PET hydrolysis effects to the purified enzyme. This work not only provides new insights into the process of chaperones in protein folding but also suggests a novel and efficient strategy for producing recombinant proteins.
研究领域:
生物化学
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环境科学
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蛋白质工程
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产品关联
DOI
PubMed
高影响因子
BIOPHARMACEUTICS & DRUG DISPOSITION
·
2022
·Vol. 43
·期 6
·247-254
·
IF 1.831
Ting Zhu,Yu Wu,Xue-Mei Li,Yu-Meng Jia,Huan Zhou,Li-Ping Jiang,Ting Tai,Qiong-Yu Mi,Jin-Zi Ji,Hong-Guang Xie
As an analog of clopidogrel and prasugrel, vicagrel is completely hydrolyzed to intermediate thiolactone metabolite 2-oxo-clopidogrel (also the precursor of active thiol metabolite H4) in human intestine, predominantly by AADAC and CES2; however, other unknown vicagrel hydrolases remain to be identified. In this study, recombinant human Raf kinase inhibitor protein (rhRKIP) and pooled human intestinal S9 (HIS9) fractions and microsome (HIM) preparations were used as the different enzyme sources; prasugrel as a probe drug for RKIP (a positive control), vicagrel as a substrate drug of interest, and the rate of the formation of thiolactone metabolites 2-oxo-clopidogrel and R95913 as metrics of hydrolase activity examined, respectively. In addition, an IC 50 value of inhibition of rhRKIP-catalyzed vicagrel hydrolysis by locostatin was measured, and five classical esterase inhibitors with distinct esterase selectivity were used to dissect the involvement of multiple hydrolases in vicagrel hydrolysis. The results showed that rhRKIP hydrolyzed vicagrel in vitro, with the values of K m , V max , and CL int measured as 20.04 ± 1.99 μM, 434.60 ± 12.46 nM/min/mg protein, and 21.69 ± 0.28 ml/min/mg protein, respectively, and that an IC 50 value of locostatin was estimated as 1.24 ± 0.04 mM for rhRKIP. In addition to locostatin, eserine and vinblastine strongly suppressed vicagrel hydrolysis in HIM. It is concluded that RKIP can catalyze the hydrolysis of vicagrel in the human intestine, and that vicagrel can be hydrolyzed by multiple hydrolases, such as RKIP, AADAC, and CES2, concomitantly.
研究领域:
药理学
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生物化学
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DOI
高影响因子
CARBOHYDRATE POLYMERS
·
2024
·
IF 10.7
Qianyu Zhang,Ranjun Yan,Yangyang Xiong,Hong Lei,Guanben Du,Antonio Pizzi,Buapan Puangsin,Xuedong Xi
Developing the most abundant cellulose resources in nature as wood adhesives is a challenging but significant work. In this study, an oxidized cellulose-hexamethylene diamine-urea (OCHU) resin adhesive, of high bonding performance, and excellent water resistance has been prepared. The cellulose was firstly enzymatically hydrolyzed to increase its solubility and expose more hydroxyl groups, then oxidized by sodium periodate (NaIO 4 ) to generate a biological compound being rich of aldehyde groups. The OCHU adhesive was prepared by crosslinking oxidized cellulose (OC) with a synthetic reactive polyurea (HU) polymer, which was formed through the deamidation reaction between hexamethylenediamine (H) and urea (U). This adhesive exhibited a 24 h cold water soaking strength of 1.61 MPa, 3 h hot water (63 °C) strength of 1.05 MPa and a dry strength of 1.71 MPa. Compared to oxidized cellulose-hexamethylenediamine (OCH) adhesive, a significant increase of 60 % in this adhesive's wet strength. The Schiff base and addition reactions in the preparation were confirmed by XPS, solid-state NMR, and FTIR. This cellulose-based wood adhesive has great market competitiveness for mass production and application in the plywood industry, due to its excellent bonding properties.
研究领域:
材料科学
·
林业工程
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF BIOLOGICAL CHEMISTRY
·
2016
·Vol. 292
·期 5
·1666-1678
·
IF 4.258
Zhen Qin,Shaoqing Yang,Liming Zhao,Xin You,Qiaojuan Yan,Zhengqiang Jiang
Carbohydrates are complex macromolecules in biological metabolism. Enzymatic synthesis of carbohydrates is recognized as a powerful tool to overcome the problems associated with large scale synthesis of carbohydrates. Novel enzymes with significant transglycosylation ability are still in great demand in glycobiology studies. Here we report a novel glycoside hydrolase family 16 “elongating” β-transglycosylase from Paecilomyces thermophila (PtBgt16A), which efficiently catalyzes the synthesis of higher polymeric oligosaccharides using β-1,3/1,4-oligosaccharides as donor/acceptor substrates. Further structural information reveals that PtBgt16A has a binding pocket around the −1 subsite. The catalytic mechanism of PtBgt16A is partly similar to an exo-glycoside hydrolase, which cleaves the substrate from the non-reducing end one by one. However, PtBgt16A releases the reducing end product and uses the remainder glucosyl as a transglycosylation donor. This catalytic mechanism has similarity with the catalytic mode of amylosucrase, which catalyzes the transglycosylation products gradually extend by one glucose unit. PtBgt16A thus has the potential to be a tool enzyme for the enzymatic synthesis of new β-oligosaccharides and glycoconjugates.
研究领域:
生物化学
·
糖生物学
·
酶学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2025
·
IF 6.2
Shanshan Song,Xina Yu,Changming Xie,Zhanhua Li,Ying Zhang,Yan Liu,Zhongjue Qiu,Tiantian Wang,Hongna Su,Hui Huang,Pei Luo
Panaxynol (PA), a bioactive compound found in carrots and other Apiaceae plants, holds promise for managing diabetes-related vascular complications. This study investigated the long-term effects of PA on diabetes-induced vascular calcification (DVC) in db/db mice for up to 9 weeks. The impact of PA on DVC was evaluated both in vivo and in vitro using immunohistochemical staining, Western blotting, micro-computed tomography, and other analytical techniques. Results demonstrated that PA significantly attenuated arterial calcification and collagen hyperplasia in the aorta. Additionally, PA elevated levels of the lipid signaling molecule 14,15-EET, which preserved the vascular structure and function by reducing inflammatory macrophage infiltration in perivascular adipose tissue. This protective effect was partially attributed to the PA-mediated inhibition of soluble epoxide hydrolase (sEH) through Sirt6 activation. These findings highlight PA as a potential natural therapeutic strategy for mitigating diabetic vascular complications and offer an alternative to conventional pharmacological approaches.
研究领域:
生物学
·
医学
·
药理学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2018
·
IF 3.909
Shu-Ping Zou,Xiu-Ling Xuan,Zhi-Jian Wang,Yu-Guo Zheng
Epoxide hydrolase-mediated biocatalysis has a great prospective in the biosynthesis of optically pure epoxides. The present work targets toward the thermo-stabilization of epoxide hydrolase by covalent conjugation with polysaccharide. An epoxide hydrolase from Agrobacterium radiobacter (ArEH) was modified by covalent coupling to seven oxidized polysaccharides with different chemical structures and characteristics. Among all conjugates, ArEH with ficoll exhibited both the highest specific activity (494 U/mg) and half-life at 60 °C ( t 1/2 = 183 min). The conjugated enzyme also displayed wider optimum pH and temperature ranges, higher catalytic number ( k cat ), and catalytic efficiency ( k cat / K m ) as compared to its native counterpart. The conjugation significantly decreased the enthalpy of activation (Δ H *), free energy of transition state binding (Δ G * E − T ), and free energy of activation (Δ G *) for epichlorohydrin hydrolysis. Moreover, the conjugated ArEH showed enhanced thermal stability as evidenced by its longer half-life ( t 1/2 ), lower thermal deactivation constant ( k d ), and higher D values at 50–70 °C. Furthermore, the enzymatic activity of conjugated ArEH elevated significantly by Ca 2+ and it showed increased tolerance against Co 2+ , Fe 3+ and EDTA inhibitors. Finally, fluorescence and circular dichroism spectroscopic analysis were performed to confirm the noticeable conformational changes in ArEH structure after conjugation with ficoll, which could be responsible for the observed catalytic and stability enhancement.
研究领域:
生物化学
·
酶工程
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DOI
高影响因子
TALANTA
·
2022
·
IF 6.556
Yaping Dong,Xiaojin Luo,Yiqun Liu,Cunling Yan,Haixia Li,Jicheng Lv,Li Yang,Yue Cui
In clinical practice, sera creatinine level is regarded as a crucial biomarker for the diagnosis, staging and monitoring of kidney disease. An amperometric biosensor is rapid, accurate, and cost-effective, with a portability and a simple operation. Herein, we report for the firsttime a disposable, printed amperometric biosensor for the clinical evaluation of creatinine in renal function detection. The sensor is constructed based on Prussian blue/carbon-graphite paste as the working electrode and the immobilization of creatinine amidohydrolase, creatine amidinohydrolase and sarcosine oxidase. The creatinine biosensor shows a linear detection range from 0.05 to 1.4 mM with a detection time of about 3 min. In addition, the sensor shows a high stability that can maintain above 86% of the initial activity after being stored for over 4 months. Moreover, the sensor shows almost the same results as those with the Jaffe method for measuring the real blood samples. We anticipate that the creatinine biosensor could be widely used in the medical and healthcare areas, especially for at-home testing and onsite medical examinations.
研究领域:
临床化学
·
生物医学工程
·
医疗器械
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产品关联
DOI
PubMed
高影响因子
STARCH-STARKE
·
2024
·
IF 2.6
Wenda Wang,Ning Zhang,Yifan Cui,Qiang Wang,Man Zhou,Ping Wang,Yuanyuan Yu
Enzymatic modification, compared with traditional starch (St) chemical modification methods, avoids using toxic solvents and toxic catalysts. The horseradish peroxidase (HRP), being the most commonly employed natural green initiator, has found extensive application in starch grafting modification as a substitute for chemical initiators. HRP-catalyzed grafting of starch with vinyl zwitterionic monomers has not been reported. Here, modified starch (St-PSBMA) is first synthesized by grafting zwitterionic sulfobetaine [2-(Methacryloyloxy)ethyl]dimethyl-(3-sulfopropyl) (SBMA) onto starch by HRP/Acetylacetone(ACAC) binary initiation system catalyzed. The modified starch-based hydrogel is then prepared by the repeated freeze-thaw method. Compared with native starch-based hydrogels, which bind water molecules only through hydrogen bonding, the ability of modified starch-based hydrogels to bind water molecules is enhanced by ionic solvation. The modified starch hydrogels also have excellent antiprotein adsorption and antibacterial properties due to the presence of a strong hydration layer. The modified starch-based hydrogel also has excellent sensitivity and stability when used as a sensor for detecting joint motion. The binary HRP initiating system, employed in this study instead of the traditional three-component HRP initiating system, offers a novel and promising approach for grafting modification of starch without the need for strong oxidant hydrogen peroxide.
研究领域:
生物化学
·
材料科学
·
酶工程
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产品关联
DOI
高影响因子
Journal of Functional Foods
·
2016
·
IF 3.973
Chun-Feng Guo,Di Zhao,Ya-Hong Yuan,Tian-Li Yue,Bin Liu,Jing-Yan Li
The hypocholesterolaemic effects of milk fermented by Lactobacillus casei F0822 with bile salt hydrolase activity were investigated in hamsters fed a cholesterol-rich diet. Fermented milk (FM) exhibited dose-dependent hypocholesterolaemic activity. Sterilized and low-dose FM did not significantly affect the lipid profile of hamsters, whereas high-dose FM significantly reduced the levels of serum total cholesterol (TC) and non-high-density lipoprotein cholesterol (non-HDL-C), as well as hepatic TC and esterified cholesterol (EC). The daily faecal total bile acid excretion was significantly negatively correlated with the serum TC and non-HDL-C, as well as with the hepatic TC and EC. High-dose FM significantly enhanced daily faecal total bile acid excretion, which attributed to the hydrolysis of conjugated bile acids and the binding of free bile acids by the cells of strain F0822. These data suggest that the FM may serve as a functional food and has the potential to regulate cholesterol metabolism in humans.
研究领域:
营养学
·
微生物学
·
代谢组学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）