INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
·
IF 7.7
Ruirui Li,Wei Zhuang,Yan Gao,Yahan Bai,Keke Zhang,Zhenfu Wang
Herein, UiO-66 was selected as the immobilization carrier of Rhizomucor miehei lipase (RML). After etching and hydrophobic modification, the functionalized UIO-66 (H-UIO-66-OPA) was utilized for RML immobilization and the obtained RML@H-UIO-66-OPA showed about 70 % relative activity after incubation at 60 °C, which was much better than RML (20 %). RML@H-UIO-66-OPA was used in the synthesis of 1,3-dioleic acid glycerol (1,3-DAG) and the effects of reaction conditions (temperature, enzyme addition, substrate molar ratio, and time) on 1,3-DAG yield were investigated. Under the optimal conditions (temperature of 65 °C, enzyme addition of 2 wt%, substrate molar ratio of 2:1, and reaction time of 4 h), 1,3-DAG yield reached 77 %. Finally, Box-Behnken experimental design method was utilized to optimize the process parameters for the preparation of 1,3-DAG, and the final 1,3-DAG content was 79.80 %, which demonstrated that the model could predict the relationship between the factors and be suitable for the process optimization in 1,3-DAG preparation.
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
ENZYME AND MICROBIAL TECHNOLOGY
·
2021
·
IF 3.493
Miao Tian,Shaowei Huang,Zhiyuan Wang,Junying Fu,Pengmei Lv,Changlin Miao,Tao Liu,Lingmei Yang,Wen Luo
The propeptide is a short sequence that facilitates protein folding. In this study, four highly active Rhizomucor miehei lipase (RML) mutants were obtained through saturation mutagenesis at three propeptide positions: Ser8, Pro35, and Pro47. The enzyme activities of mutants P35 N, P47 G, P47 N, and S8E/P35S/P47A observed at 40 °C, and pH 8.0 were 10.19, 7.53, 6.15, and 8.24 times of that wild-type RML, respectively. The S8E/P35S/P47A mutant showed good thermostability. After incubation at 40 °C for 1 h, 98.98 % of its initial activity remained, whereas wild-type RML retained only 78.76 %. This result indicated that the enhancement of hydrophilicity of 35- and 47- amino-acid residues could promote the interaction between the propeptide and the mature peptide and the enzyme activity and expression level. Highly conserved sites had a more significant impact on enzyme performance than did other sites, similar to the Pro35 and Pro47 mutants showed in this study. This study provides a new idea for protein modification: enzyme performance can be improved through propeptide regulation.
研究领域:
生物化学
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酶学
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蛋白质工程
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DOI
PubMed
高影响因子
ACS Sustainable Chemistry & Engineering
·
2020
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IF 7.632
Xiaohui Di,Yu Zhang,Junying Fu,Qiang Yu,Zhongming Wang,Zhenhong Yuan
Dimethyl itaconate (DI), a promising biomass-derived intermediate, cannot be synthesized from itaconic acid (IA) in organisms due to the lack of metabolic pathways. It is necessary to develop a green and high-efficiency method to synthesis DI from IA. In this study, DI was, for the first time, produced from IA in an aqueous medium by an ionic liquid-strengthened enzymatic method. We found that when pretreated by the ionic liquid, 1-butyl-3-methylimidazolium hexafluorophosphate ([bmim][PF6]), the pH tolerance range of the lipase shrunk and the hydrolysis activity of it was greatly inhibited, while its esterification activity was not. Moreover, when the [bmim][PF6]-treated immobilized Rhizomucor miehei lipase (RML, 50 mg/mL) was employed in a reaction containing IA/methanol at a molar ratio of 1:30 and methanol/water in a volume ratio of 1:10 at 30 °C after 24 h, a 64.3% yield and 73.2% selectivity of DI was obtained. The results of HPLC and GC–MS revealed that in addition to the main product DI, other side products including acetic acid, propionic acid, and itaconic acid cyclolactone were also generated, which indicates that the RML-catalyzed esterification of IA and methanol has various side reactions, such as lactonization and self-decomposition of IA.
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
3 Biotech
·
2026
·Vol. 16
·期 6
·219
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IF 3.1
Wang Yuanshan,Liu Ping,Yang Xiaobo,Xu Jiabao,Qin Haibin,Weng Chunyue,Zheng Yuguo
A robust and reusable Rhizomucor miehei lipase (RML) biocatalyst was prepared by integrating bio-imprinting with resin immobilization. RML was co-imprinted with lauric acid and polyethylene glycol (PEG600) followed by immobilization on an amine-functionalized epoxy resin (ES-103B), resulting in AE@RML-LA+PEG600. After immobilization process optimization, the AE@RML-LA + PEG 600 was verified by CD, SEM, FTIR, and XRD analysis. AE@RML-LA+PEG600 exhibited improved thermal, pH, and organic solvent stability compared with free RML. Under optimized conditions (10 h, 62 °C, palmitic-acid-enriched palm stearin/oleic acid = 1:10 mol/mol, 10 wt% enzyme), AE@RML-LA+PEG600 achieved a maximum OPO content of 45.80%, and the proportion of palmitic acid at the sn-2 position relative to total palmitic acid ((sn-2, P)/P) was 54.59%, which was higher than that of the unimprinted enzyme (35.52% OPO). After five cycles, AE@RML-LA+PEG600 maintained 69.52% of its original catalytic activity, indicating its excellent operational stability. This work provides a promising imprinting-immobilization strategy for preparing durable lipases for industrial OPO production.
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DOI
PubMed
高影响因子
Catalysts
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2018
·Vol. 8
·期 3
·96
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IF 3.465
Miaad Adnan,Kai Li,Li Xu,Yunjun Yan
This study presents a one-step encapsulation method for synthesizing X-shaped zeolitic imidazolate frameworks (ZIF-8) and immobilizingRhizomucor mieheilipase (RML). We proved that the morphological structure of ZIF-8 had changed after immobilization with enhanced characterization using a field-emission scanning electron microscope, an energy-dispersive spectrometer, a transmission electron microscope, a Fourier transform infrared spectrometer, and powder X-ray diffraction. The surface area and pore size of the carrier were investigated before and after immobilization using Brunauer–Emmett–Teller and Barrett–Joyner–Halenda methods, respectively. RML@ZIF-8 exhibited high recovery activity of up to 2632%, representing a 26-fold increase in its free lipase. Encapsulated RML was used for biodiesel production from soybean oil in an isooctane system with a conversion yield of 95.6% under optimum conditions. The resulting reusability of the immobilized enzyme indicated no substantial decline in the conversion yield, which remained at 84.7% of the initial activity after 10 cycles. The stability and high performance of the immobilized enzyme are attributed to the harmony between RML and ZIF-8 based on the easy synthesis of ZIF-8 and the short time required to immobilize RML.Keywords:X-shape ZIF-8;Rhizomucor mieheilipase (RML);encapsulation technique;biodiesel yield;soybean oil
研究领域:
生物化学
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材料科学
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可再生能源
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DOI
高影响因子
PROCESS BIOCHEMISTRY
·
2026
·
IF 4.4
Haotian Ma,Peiting Li,Yuequn Liu,Xiuxia Wang,Keke Zhang,Wei Zhuang,Zhenfu Wang
Four fatty acids (lauric acid (LA), myristic acid, palmitic acid, and stearic acid were employed as the template molecules to imprint Rhizomucor miehei lipase (RML) for the synthesis of 1,3-dioleoyl-2-palmitoylglycerol (OPO) by acidolysis reaction. Results indicated that the catalytic activity of bio-imprinted RML was higher than non-imprinted RML but decreased as the aliphatic chain length of fatty acids increased. The mechanism was studied through molecular dynamics simulation, and we found that fatty acids-imprinting affected the whole conformation, lid domain, and active site of RML, thus resulting in the diverse effects of fatty acids with different aliphatic chain lengths on the substrate accessibility, catalytic triad, and the stability of catalytic intermediate of bio-imprinted RML. Finally, LA-imprinted RML was utilized to catalyze OPO synthesis. Upon the optimal reaction conditions, C52 content and ( sn -2, P)/P value of OPO product reached 58% and 65%, respectively. This study would provide structure-function insights for designing efficient bio-catalysts and contribute to the development of bio-catalysts for food industry.
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高影响因子
FOOD CHEMISTRY
·
2018
·
IF 4.946
Nanjing Zhong,Weilin Chen,Liyan Liu,Hongxiao Chen
In this study, mesoporous silica SBA-15 was modified by organic functional groups through silanization. Series of organosilane compounds were grafted onto the SBA-15, and the obtained functionalized carriers were then used to immobilize the lipase from Rhizomucor miehei (RML). The enzymatic properties of the obtained immobilized RML samples were evaluated, and the catalytic efficiencies in glycerolysis of triacylglycerols (TAG) reaction were studied. Compared with the parent SBA-15 immobilized RML, the organic modification gave a maximum improvement of enzymatic activity from 200.00 to 13211.11 U/g; in addition, TAG conversion and diacylglycerols (DAG) content increased from 21.28 to 84.24% and 15.45 to 59.03% respectively. The organic modification also decreased the sensitivity of immobilized RML in extreme pH values and increased their thermostability.
研究领域:
生物化学
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酶工程
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产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
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2022
·
IF 11.889
Miao Tian,Lingmei Yang,Pengmei Lv,Zhiyuan Wang,Junying Fu,Changlin Miao,Zhibing Li,Lianhua Li,Tao Liu,Wenyi Du,Wen Luo
Exploiting highly active and methanol-resistant lipase is of great significance for biodiesel production. A semi-rational directed evolution method combined with N-glycosylation is reported, and all mutants exhibiting higher catalytic activity and methanol tolerance than the wild type (WT). Mutant N267 retained 64% activity after incubation in 50% methanol for 8 h, which was 48% greater than that of WT. The catalytic activity of mutants N267 and N167 was 30- and 71- fold higher than that of WT. Molecular dynamics simulations of N267 showed that the formation of new strong hydrogen bonds between glycan and the protein stabilized the structure of lipase and improved its methanol tolerance. N267 achieved biodiesel yields of 99.33% (colza oil) and 81.70% (waste soybean oil) for 24 h, which was much higher than WT (51.6% for rapeseed oil and 44.73% for wasted soybean oil). The engineered ProRML mutant has high potential for commercial biodiesel production.
研究领域:
生物化学
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生物技术
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酶工程
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可再生能源
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DOI
高影响因子
Food Bioscience
·
2026
·
IF 6.2
Yiyi Lin,Xiansi Gao,Zheling Zeng,Di Kuang,Kai Fang,Tianyi Song,Ping Yu,Deming Gong,Cheng Zeng,Xianghui Yan
Medium- and long-chain triglycerides (MLCT) are a novel type of functional lipids that exhibit higher bioavailability when long-chain polyunsaturated fatty acids are esterified at the sn-2 position of the glycerol backbone. Enzyme-catalyzed synthesis of MLCT offers significant advantages, including environmental benignity, exceptional catalytic efficiency, and negligible by-product formation. Nevertheless, the inherent limitations of free enzymes in terms of recyclability and industrial implementation, remain a challenge. In this study, the hydrophobicity of the amino resin support LX-1000HAA (HAA) was modified via grafting with dodecenyl succinic anhydride (DDSA). The lipase RM was then immobilized onto the HAA-DDSA carrier via physical adsorption. The activity of the resulting immobilized lipase HAA-DDSA-RM was 1.74 times that of Lipozyme RM C and 2.13 times that of HAA-RM. Analysis of the adsorption kinetics and isotherms for lipase adsorption onto HAA-DDSA revealed that the adsorption behavior conformed to both the pseudo-second-order kinetic model and the Freundlich isotherm model. HAA-DDSA-RM was then employed in the preparation of MLCT in the reaction system comprising algal oil and capric acid. The DHA content at the sn-2 position reached 73.40% in the final product. Moreover, HAA-DDSA-RM demonstrated outstanding thermal stability, good storage stability, and superior recyclability, with 96% of its initial activity after 10 successive operational cycles. This study may provide a new approach for environmentally sustainable production of functional lipids by developing a robust immobilized lipase system.
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高影响因子
FOOD CHEMISTRY
·
2022
·
IF 9.231
Michael Kidane Ghide,Kai Li,Jianhua Wang,Saadiah A. Abdulmalek,Yunjun Yan
Nowadays, breast milk is considered as the ideal food for infants owing to the most common oleic acid-palmitic acid-oleic acid (OA-PA-OA) fatty acid distribution of the human milk fat (HMF). This study reports the synthesis of 1,3-dioleoyl-2-palmotoylglycerol (OPO)-rich human milk fat substitutes in a two-step enzymatic acidolysis reaction with Rhizomucor miehei lipase (RML) immobilized on magnetic multi-walled carbon nanotubes(mMWCNTs). The immobilized RML (RML-mMWCNTs) showed better thermal and pH stability, convenient recovery and reusability than the free soluble form. Under optimized reaction conditions (1:8 tripalmitin (PPP)/OA, 10%wt. enzyme, 50 °C, 5 h), PA content at the sn- 2 position and OA incorporation at the sn -1,3 positions reached 93.46% and 59.54%, respectively. Comparison tests have also showed that RML-mMWCNTs has better catalytic activity and reusability than the commercial lipase Lipozyme RM IM. The results suggest that RML-mMWCNTs is a promising biocatalyst for the synthesis of OPO-rich TAGs with potential use in infant formulas.
研究领域:
生物化学
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食品科学
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纳米技术
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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）