Biomolecules
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2020
·Vol. 10
·期 2
·231
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IF 4.082
Rabia Durrani,Faez Iqbal Khan,Shahid Ali,Yonghua Wang,Bo Yang
Phospholipase B (EC 3.1.1.5) are a distinctive group of enzymes that catalyzes the hydrolysis of fatty acids esterified at thesn-1andsn-2positions forming free fatty acids and lysophospholipids. The structural information and catalytic mechanism of phospholipase B are still not clear. Herein, we reported a putative phospholipase B (TmPLB1) fromTalaromyces marneffeiGD-0079 synthesized by genome mining library. The gene (TmPlb1) was expressed and the TmPLB1 was purified usingE. colishuffle T7 expression system. The putative TmPLB1 was purified by affinity chromatography with a yield of 13.5%. The TmPLB1 showed optimum activity at 35 °C and pH 7.0. The TmPLB1 showed enzymatic activity using Lecithin (soybean > 98% pure), and the hydrolysis of TmPLB1 by31P NMR showed phosphatidylcholine (PC) as a major phospholipid along with lyso-phospholipids (1-LPC and 2-LPC) and some minor phospholipids. The molecular modeling studies indicate that its active site pocket contains Ser125, Asp183 and His215 as the catalytic triad. The structure dynamics and simulations results explained the conformational changes associated with different environmental conditions. This is the first report on biochemical characterization and structure dynamics of TmPLB1 enzyme. The present study could be helpful to utilize TmPLB1 in food industry for the determination of food components containing phosphorus. Additionally, such enzyme could also be useful in Industry for the modifications of phospholipids.Keywords:free fatty acids (FFAs);NMR (nuclear magnetic resonance);phospholipase B;Talaromyces marneffei;affinity chromatography
研究领域:
生物化学
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分子生物学
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食品科学
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DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2014
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IF 2.966
Chuan-Yang Shang,Wei-Xun Li,Rui-Feng Zhang
ZnO nanowires were successfully introduced into a macroporous SiO 2 by in situ hydrothermal growth in 3D pores. The obtained composites were characterized by SEM and XRD, and used as supports to immobilize Candida antarctica lipase B (CALB) through adsorption. The high specific surface area (233 m 2 /g) and strong electrostatic interaction resulted that the average loading amount of the composite supports (196.8 mg/g) was 3–4 times of that of macroporous SiO 2 and approximate to that of a silica-based mesoporous material. Both adsorption capacity and the activity of the CALB immobilized on the composite supports almost kept unchanged as the samples were soaked in buffer solution for 48 h. The chiral resolution of 2-octanol was catalyzed by immobilized CALB. A maximum molar conversion of 49.1% was achieved with 99% enantiomeric excess of ( R )-2-octanol acetate under the optimal condition: a reaction using 1.0 mol/L ( R , S )-2-octanol, 2.0 mol/L vinyl acetate and 4.0 wt.% water content at 60 °C for 8 h. After fifteen recycles the immobilized lipase could retain 96.9% of relative activity and 93.8% of relative enantioselectivity .
研究领域:
生物化学
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材料科学
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催化工程
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DOI
PubMed
高影响因子
RSC Advances
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2017
·Vol. 7
·期 21
·12518-12523
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IF 3.108
Pengfei Zhou,Xuping Wang,Bo Yang,Frank Hollmann,Yonghua Wang
Epoxides are important synthetic intermediates for the synthesis of a broad range of industrial products. This study presents a promising solution to the current limitation of enzyme instability. By using simple deep eutectic solvents such as choline chloride/sorbitol, significant stabilization of the biocatalyst has been achieved leading to more robust reactions while using environmentally more acceptable solvents as compared to ionic liquids.
研究领域:
生物化学
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绿色化学
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DOI
高影响因子
JOURNAL OF SOL-GEL SCIENCE AND TECHNOLOGY
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2022
·Vol. 102
·期 2
·400-411
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IF 2.606
Yue Wenda,Liang Jun,Wang Honghai,Zhang Yu,Li Chunli,Su Weiyi
In the entrapment of enzymes by sol-gel method, the special procedures for obtaining the unique structure of xerogel are sometimes destructive to enzymes, while the internal structure of xerogels is easily broken. To solve the above problems, we present an economical, bio-friendly procedure for the entrapment of Candida antarctica lipase B in silica xerogel. In this procedure, an encapsulation-structured enzyme-carrying silica xerogel was produced by controlling the evaporation rate of the solvent at ambient pressure during drying, while tetramethoxysilane and methyltrimethoxysilane were used as the co-precursors. The xerogel synthesis was modified and optimized for improving the stability and reusability of the enzyme, while the xerogel characteristic structure was preserved. Moreover, the effects of the co-precursors amount on the mechanical strength, pore structure, hydrophobicity, enzyme loading, and enzyme stability of the enzyme-carrying silica xerogels were systematically investigated. And the best performance was observed at the co-precursors content of 30.28 wt%. Finally, the catalytic performance of the enzyme-carrying silica xerogels on the transesterification of n -butanol with ethyl acetate was demonstrated.
Graphical abstract
研究领域:
生物化学
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材料科学
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DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2023
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IF 3.4
Liwen Shu,Xin Zheng,Shuhua Qi,Shuizhi Lin,Yinghua Lu,Chuanyi Yao,Xueping Ling
Docosahexaenoic acid (DHA) enriched with phospholipids (PLs) (DHA-PLs) is a type of structured PL with good physicochemical and nutritional properties. Compared to PLs and DHA, DHA-PLs has higher bioavailability and structural stability and many nutritional benefits. To improve the enzymatic synthesis of DHA-PLs, this study investigated the preparation of phosphatidylcholine (PC) enriched with DHA (DHA-PC) via enzymatic transesterification of algal oil, which is rich in DHA-triglycerides, using immobilized Candida antarctica lipase B (CALB). The optimized reaction system incorporated 31.2% DHA into the acyl chain of PC and converted 43.6% PC to DHA-PC within 72 h at 50 °C, 1:8 PC: algal oil mass ratio, 25% enzyme load (based on total substrate mass), and 0.02 g/mL molecular sieve concentration. Consequently, the side reactions of PC hydrolysis were effectively suppressed and products with high PC content (74.8%) were produced. Molecular structure analysis showed that exogenous DHA was specifically incorporated into the sn-1 site of the PC by immobilized CALB. Furthermore, the evaluation of reusability with eight cycles showed that the immobilized CALB had good operational stability in the present reaction system. Collectively, this study demonstrated the applicability of immobilized CALB as a biocatalyst for synthesizing DHA-PC and provided an improved enzyme-catalyzed method for future DHA-PL synthesis.
研究领域:
食品科学
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生物化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
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