BIOPROCESS AND BIOSYSTEMS ENGINEERING
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2022
·Vol. 46
·期 2
·171-181
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IF 3.434
Zhang Yin-Jun,Wei Li-Tian,Zhou Ming-Peng,Wei Chun,Yu Xin-Jun
( R )-N-(2,6-dimethylphenyl) aminopropionic acid methyl ester (( R )-DMPM) is an important chiral intermediate of the fungicide N-(2,6-Dimethylphenyl)-N-(methoxyacetyl)-alanine methyl ester (( R )-Metalaxyl). In this study, (1) D3520 (macroporous acrylic anion resin), selected from the ten resins, was used to immobilize the esterase from Pseudochrobactrum asaccharolyticum WZZ003 (PAE07) for resoluting the ( R,S )-DMPM to obtain ( R )-DMPM. (2) Up to 20 g/L PAE07 could be immobilized onto D3520 with a high enzymatic activity of 32.4 U/g. Moreover, the Km and Vmax values of 19.1 mM and 2.8 mM/min for D3520-immobilized PAE07 indicated its high activity and stereoselectivity. (3) The optimal temperature and pH for the immobilized PAE07 were 40 ℃ and 8.0, and substrate concentration was up to 0.35 M. After 15 h reaction, the conversion rate from ( R,S )-DMPM to ( R )-DMPM was 48.0% and the e.e.p and E values were 99.5% and 1393.0, respectively. In scale-up resolution, 200 g/L substrate and 12.5 g immobilized esterase PAE07 condition, a conversion rate from substrate to product of 48.1% and a product e.e.p of 98% were obtained within 12 h, with the activity of immobilized PAE07 retained 80.2% after 5 cycles of reactions. These results indicated that the D3520-immobilized esterase PAE07 had great potential for enzymatic resolution of ( R,S )-DMPM to prepare ( R )-Metalaxyl.
研究领域:
生物化学
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酶工程
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有机合成
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DOI
PubMed
高影响因子
Catalysts
·
2024
·Vol. 14
·期 5
·304
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IF 3.8
Hong Zhang,Xin Zhang,Lei Wang,Bo Wang,Xu Zeng,Bo Ren
The immobilization of enzymes is an important strategy to improve their stability and reusability. Enzyme immobilization technology has broad application prospects in biotechnology, biochemistry, environmental remediation, and other fields. In this study, composites of chitosan (CS) and sodium alginate (SA) with Cu2+forming a double-network crosslinked structure of hydrogels were prepared and used for the immobilization of laccase. Fourier infrared spectroscopy, scanning electron microscopy, and X-ray photoelectron spectroscopy tests revealed that laccase molecules were immobilized on the composite hydrogel surface by a covalent bonding method. Compared to free laccase, the pH, temperature, and storage stability of the immobilized laccase were markedly improved. In addition, the immobilized laccase could be easily separated from the reaction system and reused, and it maintained 81.6% of its initial viability after six cycles of use. Bisphenol A (BPA) in polluted water was efficiently degraded using immobilized laccase, and the factors affecting the degradation efficiency were analyzed. Under the optimal conditions, the BPA removal was greater than 82%, and the addition of a small amount of ABTS had a significant effect on BPA degradation, with a removal rate of up to 99.1%. Experimental results indicated that immobilized laccases had enormous potential in actual industrial applications.
研究领域:
生物技术
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环境科学
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酶工程
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DOI
高影响因子
RESEARCH ON CHEMICAL INTERMEDIATES
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2024
·Vol. 50
·期 7
·3153-3177
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IF 2.8
Hao Xiaohong,Zheng Xiangsheng,Zhang Qi,Da Xiongzhi,Wang Yan
The main objective of this study was to produce biodiesel utilizing Fe 3 O 4 @ZIF-8-CRL as biocatalyst, employing high acid value waste oil. Candida Rugosa lipase (CRL) was immobilized onto a magnetic ZIF-8 organometallic framework through physical adsorption, resulting in significantly enhanced stability and chemical compatibility. The study extensively investigated the impacts of enzyme dosage, alcohol-to-oil ratio, ultrasonic power, temperature, time, and the addition of deep eutectic solvents on the conversion of fatty acid ethyl esters. The findings revealed that under the conditions of biocatalyst addition of 5.7 wt.%, alcohol–oil ratio of 6:1 mmol/g, ultrasonic power of 87.297 W, reaction temperature of 40 °C, and reaction time of 2 h, the innovative biocatalyst facilitated a conversion efficiency of fatty acid ethyl esters reaching 82.06 ± 1.63%, with a retention rate of 74.52% after three cycles. Additionally, characterization of the biocatalyst using a vibrating sample magnetometer and scanning electron microscopy demonstrated its favorable magnetic recovery potential and morphology. Coupled with its straightforward and efficient synthesis, this biocatalyst proves to be competitive in the catalysis of biodiesel preparation.
研究领域:
生物化学
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能源科学
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催化技术
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DOI
高影响因子
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
·
2020
·
IF 1.992
Jin Xu,Xuejia Shi,Xiaomeng Zhang,Zhenzhong Wang,Wei Xiao,Linguo Zhao
To efficiently recycle GH78 thermostable rhamnosidase (TpeRha) and easily separate it from the reaction mixture and furtherly improve the enzyme properties, the magnetic particle Fe 3 O 4 -SiO 2 -NH 2 -Cellu-ZIF8 (FSNcZ8) was prepared by modifying Fe 3 O 4 -NH 2 with tetraethyl silicate (TEOS), microcrystalline cellulose and zinc nitrate hexahydrate. FSNcZ8 displayed better magnetic stability and higher-temperature stability than unmodified Fe 3 O 4 -NH 2 (FN), and it was used to adsorb and immobilize TpeRha from Thermotoga petrophilea 13995. As for properties, FSNcZ8-TpeRha showed optimal reaction temperature and pH of 90°C and 5.0, while its highest activity approached 714 U/g. In addition, FSNcZ8-TpeRha had better higher-temperature stability than FN. After incubation at 80°C for 3 h, the residual enzyme activities of FSNcZ8-TpeRha, FN-TpeRha and free enzyme were 93.5%, 63.32%, and 62.77%, respectively. The organic solvent tolerance and the monosaccharides tolerance of FSNcZ8-TpeRha, compared with free TpeRha, were greatly improved. Using naringin (1 mmol/l) as the substrate, the optimal conversion conditions were as follows: FSNcZ8-TpeRha concentration was 6 U/ml; induction temperature was 80°C; the pH was 5.5; induction time was 30 min, and the yield of products was the same as free enzyme. After repeating the reaction 10 times, the conversion of naringin remained above 80%, showing great improvement of the catalytic efficiency and repeated utilization of the immobilized α-L-rhamnosidase.
研究领域:
生物化学
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酶工程
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材料科学
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DOI
PubMed
高影响因子
Foods
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2023
·Vol. 12
·期 12
·2424
·
IF 5.2
Yang Zhao,Xinrui Li,Shuo Guo,Jingwen Xu,Yan Cui,Mingzhu Zheng,Jingsheng Liu
Xylanases are the preferred enzymes for the extracting of oligosaccharides from wheat bran. However, free xylanases have poor stability and are difficult to reuse, which limit their industrial application. In the present study, we covalently immobilized free maleic anhydride-modified xylanase (FMA-XY) to improve its reusability and stability. The immobilized maleic anhydride-modified xylanase (IMA-XY) exhibited better stability compared with the free enzyme. After six repeated uses, 52.24% of the activity of the immobilized enzyme remained. The wheat bran oligosaccharides extracted using IMA-XY were mainly xylopentoses, xylohexoses, and xyloheptoses, which were the β-configurational units and α-configurational units of xylose. The oligosaccharides also exhibited good antioxidant properties. The results indicated that FMA-XY can easily be recycled and can remain stable after immobilization; therefore, it has good prospects for future industrial applications.Keywords:immobilized enzymes;kinetics;thermodynamics;wheat bran;oligosaccharide
研究领域:
生物化学
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酶学
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食品科学
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热力学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
·
IF 7.7
Wenhui Li,Yuqiu Wu,Si Long,Zhi Chen,Liangzhi Li,Xin Ju
The cascade of sugar isomerases is one of the most practical methods for producing rare sugars, and enzyme immobilization endows it with high economic efficiency, operational convenience and reusability. However, the most employed cross-linker glutaraldehyde (GA) has the disadvantages of enzyme deactivation and limitation of substrate binding. Herein, three compounds, glyoxal, GA, and 2,5-furandicarboxaldehyde (DFF) were evaluated within a previously developed cascade comprising ribose-5-phosphate isomerase and D-tagatose-3-epimerase to prepare D-ribulose form D-xylose. Analyses of surface morphology, element and chemical bond revealed that all compounds effectively cross-linked the isomerases. High concentration of the cross-linkers was generally beneficial for binding protein and preventing enzyme leak during reusing cycles. Glyoxal performed the highest immobilization rate, though it hadn't been employed as a cross-linker for enzyme immobilization. DFF mediated cross-linking revealed the highest activity recovery, substrate conversion and residual activity after reusing cycles, suggesting better biocompatibility than glyoxal and GA. After 8 rounds of recycling, the residual activity of enzyme immobilized by DFF was 61.4 %, ∼30 % higher than that of GA. This study proved a potential alternative cross-linker DFF for the immobilization of enzyme cascade with high activity recovery and reusability, which could promote the efficient production of high value-added products from biomass monosaccharides.
研究领域:
生物化学
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酶工程
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DOI
PubMed
高影响因子
SCIENCE OF THE TOTAL ENVIRONMENT
·
2021
·
IF 7.963
Kai Sun,Dan Hong,Jie Liu,Abdul Latif,Shunyao Li,Gang Chu,Wenxiu Qin,Youbin Si
The potential application of fungal laccase-assisted bioremediation for estrogen elimination has caused tremendous attention. Herein, naturally-occurring Trametes versicolor laccase ( Tv lac) was capable of significantly removing 17 β -estradiol (E2) and 17 α -ethynylestradiol (EE2) at pH 5. A lower (or higher) pH caused decline in the velocity constants ( k prcs ) by obstructing the single-electron oxidation of estrogen at Tv lac T1-Cu site (or intercepting the intramolecular-electron transfer between Tv lac T1-Cu and T2/T3-Cu sites). Humic acid (HA) and natural phenolic compounds (PCs) are omnipresent in water and can influence the conversion kinetics of estrogen in Tv lac-triggered reactions. Compared with HA-free, the k prcs values of E2 and EE2 in presence of HA respectively reduced 58.61%–83.72% and 69.72%–95.62% at different pH levels. Additionally, the k prcs values for estrogen were also hampered obviously by O -dihydroxyphenol structure of model PCs, owning to the created monomeric/polymeric O -quinones inverted estrogen phenoxy radicals. The generated carbon‑carbon/oxygen self-linking aggregates during Tv lac-mediated estrogen oxidative coupling were characterized, and the polymerization mechanisms were speculated. A fluid-bed reactor with cross-linked-entrapped Tv lac in Ca-alginate beads was established to treat E2 and EE2, respectively. Compared with free Tv lac, immobilized Tv lac prominently exhibited a higher pH and temperature stability. Especially, the fluid-bed reactor with immobilized Tv lac could be reused, keeping its high conversion efficiencies of E2 and EE2 in natural water, only changing from 99.2% to 73.6% and from 98.5% to 70.9% after 1 to 5 cycles, respectively. These findings present an alternative immobilized fungal laccase-based clean biotechnology for continuously handling estrogen-contaminated waterbodies at a large-scale.
研究领域:
环境科学
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生物化学
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应用微生物学
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DOI
高影响因子
BIOTECHNOLOGY LETTERS
·
2019
·Vol. 42
·期 2
·269-276
·
IF 2.154
Cai Xinyong,Zhang Mengjie,Wei Wei,Zhang Yinjun,Wang Zhao,Zheng Jianyong
A novel enzyme immobilization method employing metal–organic framework (MOF) encapsulation and macroporous resin adsorption was developed in this study. Candida antarctica lipase B (CALB) was firstly encapsulated onto metal–organic frame structures (Zeolitic imidazole framework-8, ZIF-8) and further bonded to macroporous resin by physical adsorption. Under optimized immobilization conditions, the activity of the prepared immobilized lipase (CALB-ZIF-8@D101) determined via the methyl esterification of oleic acid was 38.4 U/mg. Compared with free lipase, the immobilized lipase exhibited improved thermal and operational stability and organic solvent tolerance. These results demonstrate that the immobilization method of ZIF-8 encapsulation and macroporous resin adsorption enhanced enzyme properties at a superior level.
研究领域:
生物化学
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酶工程
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材料科学
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DOI
PubMed
高影响因子
NEW JOURNAL OF CHEMISTRY
·
2017
·Vol. 42
·期 3
·1665-1672
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IF 3.269
Lin Tan,Zhaojun Tan,Huixia Feng,Jianhui Qiu
Herein, a new strategy was developed to fabricate an oriented cellulase/chitosan/Fe3O4 composite, which possesses extremely high activity, reusability, and stability. Compared to the conventional methods, herein, cellulase was first mixed with cellulose; this allowed the active-centers of cellulase to combine with cellulose tightly. Then, the cellulase/cellulose mixture was immediately embedded into chitosan (CS) layers, which were coated with Fe3O4 nanoparticles. After hydrolyzing the cellulose, a flexible hollow structure was obtained. The activity of immobilized cellulase was still maintained over 50% even after 15 uses; this indicated that the composite we fabricated herein possessed much higher activity and reusability as compared to those reported in other studies. The promising stability and activity of the immobilized cellulase is very important for practical applications.
研究领域:
生物技术
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材料科学
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酶工程
✓ 已验证
DOI
高影响因子
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
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2013
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IF 2.823
Guihua Ruan,Zhengyi Chen,Meiping Wei,Yuhua Liu,Haiyun Li,Fuyou Du
Microwave-assisted enzymatic digestion (MAED) technique was applied for ginkgo protein digestion with both free and immobilized enzyme. Under the optimized conditions of MAED (0.01 g/mL substrate concentration of bromelain, 4500 U/g enzyme/ginkgo protein, 30 min, 300 W microwave power), a higher digestion rate (7.50%) and a significant increase in antioxidant activity (72.7 mg/g) were obtained in contrast with the conventional methods. With the optimized digestion conditions (0.625% glutaraldehyde (v/v), 0.4 mg/mL initial concentration of bromelain and 4 h of immobilization), the activity and effectiveness factor of immobilized bromelain were respectively 86 U and 81.6%. The results of ginkgo digestion by applying MAED indicated that the digestion rate of immobilized bromelain obtained by MAED method (6.41%) was comparable to that of free bromelain in the conventional digestion (8.13%). In both case with immobilized and free bromelain while applying MAED, a homogeneously abundant distribution of peptide fragments (from 7.863 Da to 5856 Da) and a few different peptide profiles were found. This report brings in conclusion that applying MAED with immobilized enzyme has the potential to obtain the highest number of antioxidant activity peptides.
研究领域:
食品科学
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生物化学
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DOI
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2016
·
IF 3.138
Hui Guo,Yi Tang,Yang Yu,Lu Xue,Jun-qing Qian
Enzyme immobilized on magnetic particles can be used as efficient recoverable biocatalysts under strong magnetic response. To enable re-use of enzyme, modified Fe 3 O 4 particles were used as carrier to immobilize α-amylase in this paper. Firstly, the surface of Fe 3 O 4 particles were coated with amino groups by direct using TEOS (tetraethoxysilane) followed by treatment with APTES (3-aminopropyltriethoxysilane) and then carboxylated by reacting it with succinic anhydride. In addition, the effect of the immobilization condition on enzyme activity recovery and immobilization efficiency were investigated. The results showed that the optimal immobilization occurred under following conditions: pH 5.5, 40 °C, enzyme concentration of 20 mg mL −1 , reaction time for 36 h. Using immobilized α-amylase as biocatalyst, the optimum pH and temperature for hydrolysis were observed to be 6.5 and 60 °C. The kinetics of hydrolysis reaction were studied using Michaelis–Menten equation. The affinity constant ( K m ) and maximum reaction rate ( v max ) of magnetic particles immobilization α-amylase (MPIA) was 0.543 mg mL −1 and 1.321 mg min −1 compared to those of 0.377 mg mL −1 and 6.859 mg min −1 of free enzyme. After immobilization, enzymatic activity, storage stability, thermo-stability, and reusability of MPIA were found superior to those of the free one. MPIA maintained 86% enzyme activity after 30 days and maintained 78% enzyme activity after recycling six times.
研究领域:
生物化学
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酶工程
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DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2022
·
IF 4.446
Jiacong Wu,Yi Wang,Juan Han,Lei Wang,Chunmei Li,Yanli Mao,Yun Wang
In this work, a mesoporous Zr-based MOF (MP-UIO-66) was first synthesized by biomineralization method using dextran as template and the resultant particles were applied in the immobilization of cellulase . Compared with conventional UIO-66, MP-UIO-66 has more mesoporous channels (4.6 nm) that allow cellulase (3.4 * 12.4 nm) to be entrapped into the channels, improving loading capacity and stability of immobilized enzyme . Specifically, the results showed that MP-UIO-66 had an excellent loading capacity (265 mg g −1 ) for cellulase, and the residual enzyme activity of PVP-cellulase@MP-UIO-66 (PCMU) was still about 83% after 6 cycles. Moreover, the suitable pore structure exposes the active center of cellulase to the external environment, which facilitates the mass transfer of the substrate. Based on Michaelis-Menten constant analysis, the substrate affinity (13.355 mg mL −1 ) and catalytic efficiency (1.311 mL mg −1 min −1 ) of PCMU were better than those of free cellulase (14.525 mg mL −1 , 0.732 mL mg −1 min −1 ).
研究领域:
生物化学
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材料科学
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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）