COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS
·
2025
·
IF 5.4
Hong-Hong Wang,Jia Liu,Juan Chen
With the increasing demand for lipase inhibitors and novel anti-obesity therapies, it is of paramount importance to screen Candida rugosa lipase (CRL) inhibitors from traditional Chinese medicines (TCMs) as anti-obesity drugs. Immobilized enzyme screening technology combined with capillary electrophoresis has become a powerful tool for the exploration of CRL inhibitors due to its easy separation characteristics and high separation efficiency. In this work, tannic acid (TA)-modified ZIF-67 was adopted as a readily available carrier for CRL immobilization. ZIF-67 was fabricated by mixing Co(NO 3 ) 2 ·6 H 2 O with 2-methylimidazole under ambient conditions. TA was subsequently introduced to chelate the surface metal ions of ZIF-67, yielding a ZIF-67/TA hybrid carrier. CRL was then immobilized onto the ZIF-67/TA via physical adsorption. The immobilization of CRL on TA-modified ZIF-67 minimally alters its conformation, enabling 98.7 % recovery of enzymatic activity. Additionally, it exhibits a high CRL loading capacity of 36.5 mg·g⁻¹ determined by the Bradford protein assay. The ZIF˗67/TA/CRL demonstrated notable improvements in thermal resistance, operational durability, recyclability, and storage performance. Its calculated Michaelis constant ( K m ) was determined to be 1.41 mM, while the half-maximal inhibitory concentration (IC 50 ) of orlistat against the system was measured to be 19.74 µM. Finally, capillary electrophoresis was employed to evaluate CRL inhibitions across 18 TCMs, identifying Schisandrae Chinensis Fructus as the most potent CRL inhibitor. This investigation not only presents an innovative strategy for discovering lipase inhibitors and prospective anti-obesity agents, but also underscores the therapeutic potential of TCMs in metabolic disorder intervention.
研究领域:
生物化学
·
酶学
·
药学
·
纳米技术
·
药物发现
·
中医药学
·
生物催化
✓ 已验证
产品关联
DOI
高影响因子
BIORESOURCE TECHNOLOGY
·
2024
·
IF 9.7
Zhang Kai,Xin Jiaying,Lu Xuechun
This study established a Candida rugosa lipase (CRL) system to catalyze triolein and ethyl ferulate interesterification. The products were identified, and the binding mode between the substrates and CRL was predicted through molecular docking. Three methods for preparing CRL-AuNPs were proposed and characterized. It was found that the addition of 40 mL of 15 nm gold nanoparticles increased the CRL activity from 3.05 U/mg to 4.75 U/mg, but the hybridization efficiency was only 32.7 %. By using 4 mL of 0.1 mg/mL chloroauric acid, the hybridization efficiency was improved to 50.7 %, but the enzyme activity was sharply decreased. However, when the molar ratio of Mb to HAuCl 4 was 0.2, the hybridization efficiency increased to 71.8 %, and the CRL activity was also enhanced to 5.98 U/mg. Under optimal conditions, the enzyme activity of CRL-AuNPs③ was maintained at 95 % after 6 repetitions and 85.6 % after 30 days at room temperature.
研究领域:
生物化学
·
酶工程
·
纳米生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULES
·
2017
·Vol. 22
·期 1
·179
·
IF 2.861
Chunfang Li,Shuhui Jiang,Xinying Zhao,Hao Liang
In this paper we report a facile method for preparing co-immobilized enzyme and magnetic nanoparticles (MNPs) using metal coordinated hydrogel nanofibers.Candida rugosalipase (CRL) was selected as guest protein. For good aqueous dispersity, low price and other unique properties, citric acid-modified magnetic iron oxide nanoparticles (CA-Fe3O4NPs) have been widely used for immobilizing enzymes. As a result, the relative activity of CA-Fe3O4@Zn/AMP nanofiber-immobilized CRL increased by 8-fold at pH 10.0 and nearly 1-fold in a 50 °C water bath after 30 min, compared to free CRL. Moreover, the immobilized CRL had excellent long-term storage stability (nearly 80% releative activity after storage for 13 days). This work indicated that metal-nucleotide nanofibers could efficiently co-immobilize enzymes and MNPs simultaneously, and improve the stability of biocatalysts.Keywords:metal;nucleotide;magnetic iron oxide nanoparticles;Candida rugoselipase (CRL);immobilized enzyme
研究领域:
生物化学
·
纳米技术
·
酶工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2021
·
IF 3.978
Yumei Xiao,Yanxia Chen,Ruowei Lu,Yongchao Wang,Cuijuan Wang
A novel hierarchical magnetic zeolitic imidazolate framework-8 material (Hierarchical mZIF-8) is fabricated by using the cetyltrimethylammonium bromide (CTAB) and L -histidine (His) as templates. Comparing to usual magnetic ZIF-8 (mZIF-8), there are not only micropores (1.1 nm) but also new mesopores (23.3 nm) in hierarchical mZIF-8 material. It is the first time that Candida rugosa lipase (CRL) has been immobilized into hierarchical mZIF-8 with significantly higher enzyme loading capacity than mZIF-8. Furthermore, the temperature stability, pH stability and recycling convenience of immobilized CRL are all enhanced compared with free CRL.
研究领域:
生物化学
·
材料科学
·
酶工程
✓ 已验证
产品关联
DOI
高影响因子
NEOPLASIA
·
2020
·
IF 5.696
Qing Yu,Zhiguo Hu,Yanwen Shen,Yihan Jiang,Peichen Pan,Tingjun Hou,Zhen-Qiang Pan,Jing Huang,Yi Sun
Cullin-RING E3 ligase (CRL) is the largest family of E3 ubiquitin ligase, responsible for ubiquitylation of ∼20% of cellular proteins. CRL plays an important role in many biological processes, particularly in cancers due to abnormal activation. CRL activation requires neddylation, an enzymatic cascade transferring small ubiquitin-like protein NEDD8 to a conserved lysine residue on cullin proteins. Recent studies have validated that neddylation is an attractive anticancer target. In this study, we report the establishment of an Alpha-Screen-based high throughput screen (HTS) assay for in vitro CUL5 neddylation, and screened a library of 17,000 compounds including FDA approved drugs, natural products and synthetic drug-like small-molecule compounds. Gossypol, a natural compound derived from cotton seed, was identified as an inhibitor of cullin neddylation. Biochemical studies showed that gossypol blocked neddylation of both CUL5 and CUL1 through direct binding to SAG-CUL5 or RBX1-CUL1 complex, and CUL5-H572 plays a key role for gossypol binding. On cellular level, gossypol inhibited cullin neddylation in a variety of cancer cell lines and selectively caused accumulation of NOXA and MCL1, the substrates of CUL5 and CUL1, respectively, in multiple cancer cell lines. Combination of gossypol with specific MCL1 inhibitor synergistically suppress growth of human cancer cells. Our study revealed a previously unknown anti-cancer mechanism of gossypol with potential to develop a new class of neddylation inhibitors.
研究领域:
药理学
·
肿瘤学
·
细胞生物学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
·
2016
·
IF 2.189
Yicheng Bi,Min Yu,Haiyan Zhou,Hua Zhou,Ping Wei
In our study, oleyl oleate was synthesized through enzymatic esterification of oleic acid with oleyl alcohol using Candida rugosa lipase (CRL) immobilized on macroporous resin in a solvent-free system. CRL was immobilized flexibly on the support via aldehyde-dextran, and the effectiveness of aldehyde-dextrans, as cross-linking agents with different molecular weights, was compared and discussed. The impact of various factors, such as different molecular weights of dextran , molar ratio between oleic acid and oleyl alcohol, and the temperature and stability of enzymes, on esterification was investigated as well. Our study indicated that lipase immobilized by 20000 Da dextran yielded the best result. The optimal condition to produce oleyl oleate using immobilized CRL was found in our study and turned out to be molar ratio of 1, reaction temperature of 40 °C, and a 12-h reaction time to reach reaction equilibrium. Under the optimal reaction conditions, a high percentage yield of ester was achieved (92.6%). The immobilized CRL also reached a conversion rate of 86.7% with excellent stability even after 8 cycles.
研究领域:
生物化学
·
酶工程
✓ 已验证
产品关联
DOI
高影响因子
CHINESE JOURNAL OF CHEMICAL ENGINEERING
·
2019
·
IF 1.911
Yifeng Li,Chunyu Zhang,Yan Sun
Enzyme immobilization has attracted great attention for improving the performance of enzymes in industrial applications. This work was designed to create a new support for Candida rugosa lipase (CRL) immobilization. A porous poly(vinyl acetate–divinyl benzene) microsphere coated by a zwitterionic polymer, poly(maleic anhydride- alt -1-octadecene) and N,N -dimethylethylenediamine derivative, was developed for CRL immobilization via hydrophobic binding. The catalytic activity, reaction kinetics, stabilities and reusability of the immobilized CRL were investigated. It demonstrated the success of the zwitterionic polymer coating and subsequent CRL immobilization on the porous microsphere. The immobilized lipase (p2-MS-CRL) reached 27.6 mg·g −1 dry carrier and displayed a specific activity 1.5 times higher than free CRL. The increase of V max and decrease of K m were also observed, indicating the improvement of catalytic activity and enzyme-substrate affinity of the immobilized lipase. Besides, p2-MS-CRL exhibited significantly enhanced thermal stability and pH tolerance. The improved performance was considered due to the interfacial activation regulated by the hydrophobic interaction and stabilization effect arisen by the zwitterionic polymer coating. This study has thus proved the advantages of the zwitterionic polymer-coated porous carrier for lipase immobilization and its potential for further development in various enzyme immobilizations.
研究领域:
生物技术
·
生物化学
·
材料科学
·
酶学
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）