INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2022
·Vol. 23
·期 6
·2917
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IF 6.208
Yuhong Mao,Zhenling Cai,Chenxi Zhou,Hangzhen Lan,Xiuyun Ye
This study aimed to improve the stability and catalytic properties ofThermomyces lanuginosuslipase (TLL) adsorbed on a hydrophobic support. At the optimized conditions (pH 5 and 25 °C without any additions), the Sips isotherm model effectively fitted the equilibrium adsorption data, indicating a monolayer and the homogenous distribution of immobilized lipase molecules. To preserve the high specific activity of adsorbed lipase, the immobilized lipase (IL) with a moderate loading amount (approximately 40% surface coverage) was selected. Polyethylenimine (PEI) and chitosan (CS) were successfully applied as bridging units to in situ crosslink the immobilized lipase molecules in IL. At the low polymer concentration (0.5%,w/w) and with 1 h incubation, insignificant changes in average pore size were detected. Short-chain PEI and CS (MW ≤ 2 kDa) efficiently improved the lipase stability, i.e., the lipase loss decreased from 40% to <2%. Notably, CS performed much better than PEI in maintaining lipase activity. IL crosslinked with CS-2 kDa showed a two- to three-fold higher rate when hydrolyzing p-nitrophenyl butyrate and a two-fold increase in the catalytic efficiency in the esterification of hexanoic acid with butanol. These in situ crosslinking strategies offer good potential for modulating the catalytic properties of TLL for a specific reaction.Keywords:polyethylenimine;chitosan;anti-desorption;hydrolytic activity;esterification
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Frontiers in Bioengineering and Biotechnology
·
2020
·
IF 3.644
Bi Yanhong,Wang Zhaoyu,Tian Yaoqi,Fan Haoran,Huang Shuo,Lu Yihui,Jin Zhengyu
The immobilization of Thermomyces lanuginosus lipase on polydopamine-functionalized Fe3O4 magnetic nanoparticles (Fe3O4@PDA-TLL) as a nanobiocatalyst was successfully performed for the first time, and the Fe3O4@PDA-TLL was used for regioselective acylation of natural hyperoside with vinyl decanoate. The effects of several crucial factors, such as the reaction solvent, substrate molar ratio, temperature, and immobilized enzyme dosage, were investigated. Under optimum conditions, the reaction rate, 6″-regioselectivity, and maximum substrate conversion were as high as 12.6 mM/h, 100%, and 100%, respectively. An operational stability study demonstrated that the immobilized enzyme could maintain 90.1% of its initial maximum conversion even after reusing it five times. In addition, further investigations on the kinetic parameters, like Vmax, Km, Vmax/Km, and Ea, also revealed that the biocompatible Fe3O4@PDA could act as an alternative carrier for the immobilization of different enzymes.
研究领域:
生物化学
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纳米生物学
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酶工程
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Frontiers in Bioengineering and Biotechnology
·
2021
·
IF 5.89
Wang Zhaoyu,Li Yang,Li Mingyi,Zhang Xiaohui,Ji Qingxia,Zhao Xiaojuan,Bi Yanhong,Luo Si
The conversion of flavonoid glycosides and their analogs to their lipophilic ester derivatives was developed by nanobiocatalysts from immobilizing Thermomyces lanuginosus lipase (TLL) on polydopamine-functionalized magnetic Fe3O4 nanoparticles (Fe3O4-PDA-TLL). The behavior investigation revealed that Fe3O4-PDA-TLL exhibits a preference for long chain length fatty acids (i.e., C10 to C14) with higher reaction rates of 12.6–13.9 mM/h. Regarding the substrate specificity, Fe3O4-PDA-TLL showed good substrate spectrum and favorably functionalized the primary OH groups, suggesting that the steric hindrances impeded the secondary or phenolic hydroxyl groups of substrates into the bonding site of the active region of TLL to afford the product.
研究领域:
生物化学
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酶工程
·
有机化学
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INDUSTRIAL CROPS AND PRODUCTS
·
2023
·
IF 5.9
Xiaoyan Ji,Rui Xu,Fei Wang,Xun Li
Biodiesel is recognized as green and renewable energy with low carbon intensity and less environmental impacts. The preparation of an economical immobilized lipase becomes much urgent challenge in the enzymatic production of biodiesel. In this study, recombinant Thermomyces lanuginosus lipase (TLL) was immobilized on the surface of silica-containing materials mediated by solid-binding peptides (SBPs). The obtained immobilized biocatalysts were effective in converting the oil from Jatropha curcas into biodiesel. The appropriate attachment mode of SBPs and TLL was investigated firstly, and the results showed that the activity of TLL was not negatively affected only when SBPs were fused at the C-terminus of TLL. Secondly, the immobilization efficiency of different SBPs (CecA, EctP1, VKT) on different silica-containing materials (SiO 2 , zeolite ZSM-5, SAR-100, MCM-41 and Na-Y) was systematically studied and it was concluded that the best immobilization efficiency was achieved for TLL-EctP1 immobilized on zeolite ZSM-5 (TLL- EctP1 @ZSM-5). TLL-EctP1 @ZSM-5 was found great performance in stability of pH, temperature, elution and storage. Eventually, biodiesel yield of 93 wt% was achieved using TLL-EctP1 @ZSM-5 as a biocatalyst to catalyze J. curcas oil for 48 h, and 81% of its initial yield was retained after 7 cycles of reuse. These results highlight the application of solid-binding peptides in enzyme immobilization for biocatalysts used in preparation of biodiesel or other chemicals.
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JOURNAL OF THE AMERICAN OIL CHEMISTS SOCIETY
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2023
·
IF 2
Miao Qiao,Xin Hua,Yongjun Yuan,Xi Cao
This work attempts to achieve the enrichment of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in acylglycerols as well as biodiesel production by investigating the performance of various lipases in ethanolysis. The ethanolysis activities of five immobilized lipases in decreasing order were as follows: Lipozyme TL IM, Novozym 40086/Lipozyme RM IM, Novozym 435 and Rhizopus oryzae lipase immobilized on polypropylene (RO PP). EPA could be enriched to a certain content by Lipozyme TL IM, Novozym 40086, Lipozyme RM IM, and RO PP. The enrichment of DHA was achieved well by those four sn -1,3 regioselective lipases, and DHA contents in acylglycerols were the highest when Lipozyme TL IM and Novozym 40086 were used, reaching 2.87-fold and 2.81-fold of the initial content. However, Novozym 435 could not enrich EPA and DHA effectively. Fatty acid ethyl esters produced in this work contained abundant palmitic acid and palmitoleic acid, which could be ideal biodiesel considering their chain length. The competitive factors of five lipases towards DHA were all significantly higher than those towards EPA. When Lipozyme TL IM and Novozym 40086 achieved the maximum DHA contents, their DHA recoveries in acylglycerols reached 91.87% and 89.00%, respectively. The DHA recoveries were all significantly higher than the corresponding EPA recoveries in the reactions catalyzed by five lipases. Moreover, Lipozyme TL IM could be a potential lipase for the separation of EPA and DHA. This study may promote the full use of fish oil and broaden the application of lipases.
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Food Chemistry-X
·
2024
·
IF 6.5
Tiehan Li,Yuming Wei,Mingxia Lu,Yida Wu,Yanqun Jiang,Han Ke,Aiju Shao,Jingming Ning
Piling fermentation (PF) is crucial for Pu-erh tea aroma, yet its microbial and moist-heat impact on aroma quality is poorly understood. Solid-phase microextraction, solvent-assisted flavor evaporation, and gas chromatography–mass spectrometry were used to detected and analyses the samples of sun-green green tea, sterile PF and spontaneous PF. Microbiological action promotes the formation of stale aromas. Moist-heat action promotes the formation of plum-fragrance and sweet aroma. 20 microbial markers and 28 moist-heat markers were screened from 184 volatile components. Combining odor activity values and gas chromatography-olfactometry, 22 aroma-active compounds were screened (1,2,3-trimethoxybenzene, linalool, 1,2,4-trimethoxybenzene …), and analyzed during PF processing. Aroma omission and addition experiments verified its importance. Gallic acid addition experiments successfully verified that microorganisms are the main contributors to the synthesis of methoxybenzenes. Finally, Blastobotrys, Rasamsonia , and Thermomyces showed positive correlation with the synthesis of 1-ethyl-4-methoxybenzene, 1,2,4-trimethoxybenzene, 1,2,3-trimethoxybenzene, and 1,2-dimethoxybenzene. The formation mechanism of Pu-erh tea's aroma was clarified. Exploring microbial and moist-heat effects on Pu-erh tea volatiles and understanding the methoxybenzene formation mechanism using molecular sensory science.
研究领域:
食品科学
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微生物学
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感官科学
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化学
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Bioresources and Bioprocessing
·
2024
·Vol. 11
·期 1
·1-14
·
IF 4.3
Chen Ming,She Weina,Zhao Xin,Chen Cheng,Zhu Benwei,Sun Yun,Yao Zhong
The Thermomyces lanuginosus lipase (TLLs) was successfully immobilized within a novel hydrogel matrix through a two-step crosslinking method. TLLs were initially crosslinked through the Schiff base reaction by oxidized carboxymethyl cellulose (OCMC). The water-soluble OCMC@TLLs complex was subsequently crosslinked by carboxymethyl chitosan (CMCSH) in a microfluidic apparatus to form the CMCHS/OCMC@TLLs microspheres. The CD (Circular Dichroism, CD) and FT-IR (Fourier Transform infrared spectroscopy, FT-IR) spectra demonstrated that the crosslinking of TLLs with OCMC resulted in a less significant impact on their structure compared to that with glutaraldehyde. CMCHS/OCMC@TLLs showed decreased catalytic performance due to the mass transfer resistance, while its thermal stability was greatly improved. The CMCHS/OCMC@TLLs were used to catalyze the lauroylation of arbutin in tetrahydrofuran. After 12 h of reaction under optimal conditions, the yield of 6′-O-lauryl arbutin reached an impressive 92.12%. The prepared 6′-O-lauryl arbutin has high lipophilicity and exhibits similar tyrosinase inhibitory activity and higher antioxidant activity compared to its parent compound.
研究领域:
生物化学
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酶工程
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食品科学
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