JOURNAL OF MEMBRANE SCIENCE
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2017
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IF 6.035
Jinxin Fan,Jianquan Luo,Yinhua Wan
Enzyme purification and subsequent immobilization usually require multiple steps and consume plenty of chemicals. Traditional dipping incubation operation is lengthy and the interior of porous carrier is not fully exploited due to the diffusion barrier . To solve these problems, one-step enzyme purification and immobilization was developed based on membrane chromatography concept. A membrane adsorber was prepared by polydopamine coating on a PVDF microfiltration membrane (or on a cheap and disposable cellulose filter paper) and subsequent polyethyleneimine grafting, without any solvent usage. Laccase was selectively immobilized on such membrane adsorber (enzyme activity: 39.9±4.4 U/mL; expressed activity: 19.6±3.0%) by directly capturing it from a crude fermentation broth without pH adjustment under flow-through mode. The immobilized laccase had a high purity of 92.2%, while the membrane permeability decreased due to the adsorption of laccase and impurities (“membrane fouling”). Thanks to the dominant convective transport, the enzyme loading was high and much faster than that by dipping incubation (5 min vs. 80 min). Moreover, enzyme reloading on membrane adsorber was easily realized by reversible desorption-adsorption process. The constructed biocatalytic membrane exhibited a commendable reusability for bisphenol A (BPA) removal only driven by its gravity.
研究领域:
生物化学
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环境科学
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材料科学
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产品关联
DOI
高影响因子
JOURNAL OF ENVIRONMENTAL MANAGEMENT
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2021
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IF 6.789
Ting Yu,Hengyan Ma,Hui Zhang,Minghua Xiong,Yuan Liu,Feng Li
The fragility and high cost of enzymes represent critical challenges limiting their practical application in the removal of pesticides. Herein, an aryloxyphenoxypropionate herbicide-hydrolyzing enzyme, QpeH, was purified via one-step affinity chromatography and embedded into two types of zeolitic imidazolate frameworks (ZIFs) through biomimetic mineralization . The catalytic activity towards the herbicide quizalofop-P-ethyl, the loading capacity and efficiency of the resulting two composites, QpeH@ZIF-10 with cruciate flower-like morphology and QpeH@ZIF-8 with rhombic dodecahedral morphology, were compared. Both composites had excellent stability and reusability after 10 reuse cycles, with QpeH@ZIF-10 having a better performance. More importantly, when applied for the removal of quizalofop-P-ethyl pollution in the watermelon field, QpeH@ZIF-10 (88%) showed a remarkably improved degradation efficiency compared to QpeH@ZIF-8 (84%) despite the latter having a greater loading capacity. Finally, the use of QpeH@ZIF composites was shown to recover the bacterial community in soil. This work provides a new insight into the low-cost synthesis of nanobiocatalysts combining simple purified enzymes and metal-organic frameworks (MOFs) for the remediation of pesticide-contaminated soils.
研究领域:
环境科学
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生物技术
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DOI
高影响因子
INTERNATIONAL JOURNAL OF FOOD SCIENCE AND TECHNOLOGY
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2018
·Vol. 54
·期 2
·480-489
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IF 2.383
Dianyu Yu,Ruchun Zhang,Yuqi Wang,Dezhi Zou,Tingting Li,Honglin Tang,Lianzhou Jiang,Liqi Wang
Summary β-Carotene 15,15′-monooxygenase was isolated and purified from the intestinal mucosa of pigs, and then, it was applied to hydrolyse the pigment in soybean oil, and thus, vitamin A fortified soybean oil was obtained. The pig intestinal mucosal protein solution was purified to a specific activity of 2.487 × 10 −4 U mg −1 , maximum reaction velocity ( V max ) of 8.42 × 10 −9 mol/h and Michaelis constant (K m ) of 2.03 × 10 −5 m . The protein solution had a molecular mass of 156 kDa by gel filtration. The sodium deoxycholate concentrations, optimum pH, Tween 40 amount and enzyme amount for vitamin A production in soybean oil were determined to be 6.0 m m , 8.0, 3.0% (w/v) and 0.2 U/mL enzyme, respectively. Under these conditions, β-carotene 15,15′-monooxygenase produced 14.65 mg/L vitamin A after 20 h, with a conversion yield of β-carotene of 33.29% (w/w). Therefore, the nutrients in soybean oil were improved.
研究领域:
生物化学
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食品科学
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营养学
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产品关联
DOI
高影响因子
Food Chemistry-X
·
2023
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IF 6.1
Jialin Wu,Guangqun Lan,Na He,Laping He,Cuiqin Li,Xiao Wang,Xuefeng Zeng
A producing-fibrinolytic enzyme strain was isolated with high yield. The strain was identified as Bacillus amyloliquefaciens . B. amyloliquefaciens GUTU06 fibrinolytic enzyme was purified by acetone precipitation and reverse micelle. Acetone precipitation condition and reverse micelle condition were examined. Results showed that the total reverse micelle extraction efficiency was 64.49 % ± 1.6 %. The purification fold of the entire process reached 13.38. The optimum pH of purified enzyme is 5, and the optimum temperature is 45 °C. Fe 3+ and K + can enhance the fibrinolytic activity of the enzyme. Compared to commercial fibrinolytic enzymes such as urokinase and lumbrukinase, GUTU06 fibrinolytic enzymes have a lower pH optimal range and higher temperature stability. The molecular weight of the enzyme was approximately 28 kDa. Reverse micelle extraction with cetyl trimethylammonium bromide as a surfactant combined with acetone precipitation is suitable for separating and purifying fibrinolytic enzymes and a promising technique for obtaining active proteins.
研究领域:
生物化学
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酶学
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微生物学
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生物技术
✓ 已验证
DOI
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
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2021
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IF 13.273
Jiasheng Shi,Henan Zhang,Qian Wang,Zhengyu Duan,Li Xu,Fenghao Guo,Yunfeng Xie,Zhiyong Chen
To adapt to nature, wood forms an oriented tubular porous structure that is feasible for transporting water and nutrition. Inspired from this adaptability, we here demonstrate a universal approach to obtain rigid cryogels of aligned micro-sized tubular structures by a redox-induced cryopolymerization conducted in a freezer. In contrast to the classical directional freezing method, our method free of any special instrument leads to the aligned porous structures that are dependent on gravity due to the tubular porous structures parallel to gravity direction regardless of the sample placement direction. The versatility of our method is proven in the fabrications of a series of cryogels varying different hydrophilicity. Among the resultant samples, the three-function crosslinked cryogenic sample containing carboxyl group (polyMAA-TRIM, 40/60) was tested to have an unprecedented property integrated by high mechanical strength (9.0 MPa of compressive moduli), good hydrophilicity, and efficient purification of lysozyme (purity > 90 %) from egg white solutions only driven by gravity.
研究领域:
材料科学
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生物医学工程
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产品关联
DOI
高影响因子
TALANTA
·
2024
·
IF 5.6
Ya-Qi Zhao,Zhi-Peng Li,Shi-Chao Dong,Hao Wang,Yi-Mei Zhao,Lin-Yi Dong,Zhen-Yu Zhao,Xian-Hua Wang
The effective method for trypsin purification should be established because trypsin has important economic value. In this work, a novel and simple strategy was proposed for fabricating micron-sized magnetic Fe 3 O 4 @agarose-benzamidine beads (MABB) with benzamidine as a ligand, which can efficiently and selectively capture trypsin. The micro-sized MABB, with clear spherical core-shell structure and average particle size of 6.6 μm, showed excellent suspension ability and magnetic responsiveness in aqueous solution. The adsorption capacity and selectivity of MABB towards target trypsin were significantly better than those of non-target lysozyme. According to the Langmuir equation, the maximum adsorption capacity of MABB for trypsin was 1946 mg g −1 at 25 °C, and the adsorption should be a physical sorption process. Furthermore, the initial adsorption rate and half equilibrium time of MABB toward trypsin were 787.4 mg g −1 min −1 and 0.71 min, respectively. To prove the practicability, MABB-based magnetic solid-phase extraction (MSPE) was proposed, and the related parameters were optimized in detail to improve the purification efficiency. With Tris-HCl buffer (50 mM, 10 mM CaCl 2 , pH 8.0) as extraction buffer, Tris-HCl buffer (50 mM, 100 mM CaCl 2 , pH 8.0) as rinsing buffer, acidic eluent (0.01 M HCl, 0.5 M NaCl, pH 2.0) as eluent buffer and alkaline buffer (1 M Tris-HCl buffer, pH 10.0) as neutralization solution, the MABB-based MSPE was successfully used for trypsin purification from the viscera of grass carp ( Ctenopharyngodon idella ). The molecular weight of purified trypsin was determined as approximate 23 kDa through sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The purified trypsin was highly active from 30 °C to 60 °C, with an optimum temperature of 50 °C, and was tolerant to pH variation, exhibiting 85 % of maximum enzyme activity from pH 7.0 to 10.0. The results demonstrated that the proposed MABB-based MSPE could effectively purify trypsin and ensure the biological activity of purified trypsin. Therefore, we believe that the novel MABB could be applicable for efficient purification of trypsin from complex biological systems.
研究领域:
生物化学
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生物技术
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分析化学
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蛋白质纯化
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酶学
✓ 已验证
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）