STARCH-STARKE
·
2017
·
IF 1.837
Zixing Dong,Xiaoming Hao,Daman S. Pokhrel,Xiaoling Chen,Xiaoguang Liu,Nokuthula P. Mchunu,Kugenthiren Permaul,Suren Singh,Dandan Niu,Zhengxiang Wang
High-glucose syrup has wide applications in the feed and fermentation industries. However, during its regular production process, because of the slow hydrolysis of α-1,6-glucosidic linkages in maltodextrins by glucoamylase, yields of glucose higher than 96% are rarely achieved. To find a suitable enzyme for the saccharification process, two novel genes encoding oligo-1,6-glucosidases (OGL, EC 3.2.1.10) from Bacillus mycoides ( bmogl ) and Thermomyces lanuginosus ( tlogl ) were successfully cloned and overexpressed in Pichia pastoris GS115. At the shake-flask fermentation level, the OGL activities of the two recombinants GS115 (pPIC9K- bmogl ) and GS115 (pPIC9K- tlogl ) were 994 and 1219 U/mL, respectively; and mature enzymes around 66–68 kDa were purified for characterization. Recombinant enzyme TlOGL exhibited higher thermostability than ever reported for OGLs, whereas BmOGL was stable under acidic pH, ranging from 4.0 to 8.0. Among the substrates tested, these two recombinant enzymes efficiently hydrolyzed isomaltose and isomaltotriose, but had no activity against maltose and maltotriose. Besides these characteristics, nearly complete hydrolysis of isomaltotriose and 90% of isomaltooligosaccharides (IMOs) into glucose was also observed for them, which makes them good candidates for subsequent use in improving the yield of glucose from starch. To the best of our knowledge, this is the first report on the expression and characterization of OGLs from B. mycoides and T. lanuginosus .
研究领域:
生物化学
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分子生物学
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工业酶学
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DOI
高影响因子
RSC Advances
·
2024
·Vol. 14
·期 16
·10953-10961
·
IF 3.9
Shi-Yi Zhang,Guo-Neng Fu,Li-Hua Du,Hang Lin,Ao-Ying Zhang,Han-Jia Xie,Zhi-Kai Sheng,Miao-miao Xue,Bing-lin Yan,Yi Liu,Zhi-Xuan Ruan,Bing-Le Pan,Tong-Yao Zhou,Xi-Ping Luo
Purine nucleoside ester is one of the derivatives of purine nucleoside, which has antiviral and anticancer activities. In this work, a continuous flow synthesis of purine nucleoside esters catalyzed by lipase TL IM from Thermomyces lanuginosus was successfully achieved. Various parameters including solvent, reaction temperature, reaction time/flow rate and substrate ratio were investigated. The best yields were obtained with a continuous flow microreactor for 35 min at 50 °C with the substrate ratio of 1 : 5 (nucleosides to vinyl esters) in the solvent of tert-amyl alcohol. 12 products were efficiently synthesized with yields of 78–93%. Here we reported for the first time the use of lipase TL IM from Thermomyces lanuginosus in the synthesis of purine nucleoside esters. The significant advantages of this methodology are a green solvent and mild conditions, a simple work-up procedure and the highly reusable biocatalyst. This research provides a new technique for rapid synthesis of anticancer and antiviral nucleoside drugs and is helpful for further screening of drug activity.
研究领域:
生物催化
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药物化学
·
有机合成
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DOI
PubMed
高影响因子
JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE
·
2019
·Vol. 100
·期 4
·1426-1435
·
IF 2.422
Xihong Zhao,Fenghuan Zhao,Nanjing Zhong
BACKGROUND In this study, SBA-15 was functionalized by silane coupling reagents, then lipase from Thermomyces lanuginosus (TLL) was immobilized onto the parent and the organically modified SBA-15 for diacylglycerol (DAG) production through glycerolysis. RESULTS Diacylglycerol content of 54.77 ± 0.63%, and triacylglycerol (TAG) conversion of 77.75 ± 1.24%, were obtained from the parent SBA-15 supported TLL-mediated glycerolysis reaction in a solvent-free system. However, poor performance was unexpectedly observed when co-solvents were introduced into the reaction system. After organic modification, the functionalized SBA-15 supported TLL samples all exhibited reasonable performance, producing DAG content over 40 wt% and TAG conversion over 70 wt%. Higher DAG content, up to 59.19 ± 1.10%, was observed from the phenyl group-modified SBA-15 supported TLL. The operational stability of the immobilized TLL samples in glycerolysis was also improved after organic functionalization. The phenyl group-modified SBA-15 supported TLL showed good reusability in the present glycerolysis reaction, and 95.21 ± 4.87% of the initial glycerolysis activity remained after five cycles of reuse. CONCLUSION The organic modification of SBA-15 improved the catalytic performance of its supported TLL in glycerolysis, in terms of TAG conversion, DAG content, and reusability. © 2019 Society of Chemical Industry
研究领域:
化学工程
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生物催化
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材料科学
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
·
2022
·Vol. 23
·期 6
·2917
·
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
研究领域:
生物化学
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生物催化
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材料科学
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DOI
PubMed
高影响因子
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
·
2015
·Vol. 64
·期 1
·87-91
·
IF 1.362
Hong De Yan,Qin Li,Zhao Wang
A lipase from Thermomyces lanuginosus (Lipozyme TL IM) exhibited high enantioselectivity for kinetic resolution of (±)-menthol in organic solvent. The various reaction parameters affecting the conversion and enantioselectivity were studied. The optimum reaction conditions for the transesterification reaction were found with vinyl acetate in the solvent of methyl tert -butyl ether with a vinyl acetate:(±)-menthol molar ratio of 5:1 and an enzyme concentration of 200 g/L at 30 °C. In these conditions, (−)-menthyl acetate with 99.3% enantiomeric excess was obtained, whereas the conversion was 34.7% with the reaction time of 12 H at the substrate concentration of 0.5 M. In addition, the enzyme allowed the substrate loading to be increased up to 1.5 M without the decrease of the enantioselectivity. These results indicated that Lipozyme TL IM was a promising biocatalyst in the resolution of (±)-menthol.
研究领域:
生物催化
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酶工程
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有机化学
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立体化学
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DOI
高影响因子
Microbial Biotechnology
·
2025
·Vol. 18
·期 10
·e70254
·
IF 5.2
Donglou Wang,Gang Dai,Jiangbo He,Huiwen Si,Chunhua Liao,Wenjie Wang,Zhangxin Zuo,Shuhong Li,Xuemei Niu
Fungal pigments, particularly anthraquinones, play critical roles in food industrial applications due to their vivid hues and bioactive properties. Despite their significance, the functional mechanisms underlying pigment variation and their natural functions remain poorly understood. Here, we investigate the genetic and biochemical basis of anthraquinone‐mediated pigmentation in the thermophilic fungus Thermomyces dupontii . Through a combination of transcriptomic, biochemical and chemical analyses, we identified the flavin‐dependent halogenase gene ( hal ) as a key controller of anthraquinone chlorination, a process essential for fungal pigmentation and metabolic responses to low temperature stress. Disruption of hal abolished chlorinated anthraquinone production, reduced colony pigmentation and impaired ATP production. We characterised novel chlorinated and dichlorinated anthraquinones, highlighting chlorination's role in enhancing the structural diversity of anthraquinones and fungal pigmentation. Detailed bioassays indicated that hal ‐mediated anthraquinone chlorination contributes to fungal survival under cold stress by enhancing two distinct energy modes. This study provides new insights into the ecological significance of fungal pigmentation and opens avenues for the biotechnological exploitation of fungal pigments in food industrial and medical research. This diagram illustrates the chlorination of anthraquinones and its biological functions in Thermomyces dupontii , elucidating how chlorination‐mediated fungal pigmentation promotes energy production and stress adaptation. (1) Extracellular Fenton Chemistry: Involves superoxide anion (O 2 − ) and ferrous ion (Fe 2+ ), which are two fundamental elements for extracellular Fenton chemistry, which is strongly enhanced by polyketide‐derived emodin (5) and carviolin A (1) that facilitate the formation of Fe 2+ and O 2 − and export Fe 2+ . The extracellular Fenton chemistry can degrade organic matter to generate energy and elevate the ambient temperatures, collectively enabling fungal adaptation to low‐temperature conditions. (2) Anthraquinone Chlorination: Halogenase (Hal) in T. dupontii can introduce chlorine (Cl) to emodin and carviolin A, generating yellow monochlorinated intermediates: carviolin A‐Cl (2) and emodin‐Cl (4). These two intermediates were further fused to form a new dark purple dimeric anthraquinone. (3) Linkage to Two Energy Modes: The chlorination of anthraquinones enhances intracellular ATP production and extracellular Fenton chemistry via releasing protons and electrons from anthraquinone and Cl − , thereby contributing to fungal colour and vitality under low temperature stress. In summary, this figure illustrates how T. dupontii employs anthraquinone chlorination (driven by the hal gene) to enhance pigmentation, energy production and cold stress tolerance. This connection bridges chemical transformations with ecological adaptation, highlighting the biotechnological potential of fungal pigments in environmental ecology research.
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PubMed
高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
·
2019
·
IF 6.361
Wenfeng Zhou,Wei Zhuang,Lei Ge,Zhenfu Wang,Jinglan Wu,Huanqing Niu,Dong Liu,Chenjie Zhu,Yong Chen,Hanjie Ying
Graphene oxide (GO) with oxygen containing functional groups can be selectively modified by small biomolecules to achieve heterogeneous surface properties. To achieve a hyper-enzymatic activity, the surface functionality of GO should be tailored to the orientation adsorption of the Thermomyces lanuginosus (TL) lipase , and the active center can be covered by a relatively hydrophobic helical lid for protection. In this work, amino acids were used to interact with GO through reduction reaction, hydrophobic forces , electrostatic forces , or hydrogen bonding to alter the surface hydrophobicity and charge density. Characterization of the structure and surface properties confirmed that the GO samples decorated with phenylalanine (Phe) and glutamic acid (Glu) exhibited superior hydrophobicity than other modifications, whereas tryptophan (Trp) and cysteine (Cys) provided weaker reduction effects on GO. Moreover, the zeta potential of the samples modified by amino acids of lysine (Lys) and arginine (Arg) is higher than other modified samples. The adsorption amount of lipase on Glu-GO reached 172 mg/g and the relative enzymatic activity reached up to 200%. The thermodynamic data and the Freundlich isotherm model fitting showed that the lipase adsorption process on modified samples was spontaneous, endothermic and entropy increase.
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
PubMed
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Hao Zhou,Yongheng Shi,Lingmei Dai,Dehua Liu,Wei Du
Pyrolyzing ZIF-8 into N-doped carbon is now inspiring new potential of ZIF-8 for extensive applications. However, the exploration of such ZIF-8-derived carbon for biotechnological applications has been rarely investigated, especially in this context that imidazole derivatives are of rich functionality in life science. Herein, we reported a facile temperature-dependent pyrolysis method to achieve the effective N-doping in ZIF-8-derived macroporous carbon, which was systematically investigated for the immobilization of lipase from Thermomyces lanuginosus (TLL) in biodiesel production. MNC-600 (macroporous N-doped carbon derived at 600 °C) exhibited the best immobilization performance, with the highest specific activity of 190.6 U mg −1 , a 36.9% increase than free TLL. Moreover, MNC-600@TLL showed superior effectiveness and robustness in biodiesel production. A much faster rate of conversion and the highest biodiesel yield of 87.4% were observed. After eight successive batches of recycling, MNC-600@TLL retained 93.4% of its initial activity. XPS analysis revealed the evolution of N species during the pyrolysis of ZIF-8. It was proposed that the abundant doping of pyrrolic N species played a major role in the surprising performance of MNC-600, mainly through hydrogen bonding interactions, which simultaneously realized the activation of reactants as well as the in-situ hydrophobization of biocatalyst. This report is anticipated to pave a way for the exploration of ZIF-8-derived N-doped carbon for extensive biotechnological applications.
研究领域:
材料科学
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生物技术
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化学工程
·
催化
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产品关联
DOI
高影响因子
Catalysts
·
2022
·Vol. 12
·期 3
·339
·
IF 4.501
Li-Hua Du,Meng-Jie Yang,Yue Pan,Ling-Yan Zheng,Shi-Yi Zhang,Zhi-Kai Sheng,Ping-Feng Chen,Xi-Ping Luo
Coumarin carboxamide derivatives are important building blocks for organic synthesis and chemical biology due to their excellent biopharmaceutical properties. In this paper, we demonstrate for the first time a two-step enzymatic synthesis of coumarin carboxamide derivatives. Salicylaldehyde and dimethyl malonate were reacted to obtain coumarin carboxylate methyl derivatives, which were then reacted with various amines under the catalysis of lipase TL IM fromThermomyces lanuginosusto obtain coumarin carboxamide derivatives in continuous flow reactors. We studied various reaction parameters on the yields. The important features of this method include mild reaction conditions, a short reaction time (40 min), reduced environmental pollution, higher productivity (STY = 31.2941 g L−1h−1) and enzymes being relatively easy to obtain.Keywords:enzymatic synthesis;microreactor;coumarin carboxamide derivatives
研究领域:
有机化学
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生物催化
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药物化学
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产品关联
DOI
高影响因子
JOURNAL OF COLLOID AND INTERFACE SCIENCE
·
2016
·
IF 3.782
Ruipu Xin,Faez Iqbal Khan,Zexin Zhao,Zedong Zhang,Bo Yang,Yonghua Wang
The mechanism of lipase binding to the lipid-water interface is crucial for substrate specificity and kinetic properties. In this study, the chain-length specificity, regiospecificity and substrate specificity of Phospholipase A 1 (PLA 1 ) and its parent enzyme Thermomyces lanuginosus lipase (TLL) have been investigated using a classical emulsion system. The results show that both PLA 1 and TLL are 1,3-regioselective lipases. Additionally, the hydrolytic activity of PLA 1 is comparatively lower on short-chain triacylglyceride (TAG) and higher on phosphatidylcholine (PC) than the hydrolytic activity of TLL. Further, the results obtained with monolayer film techniques demonstrate that the C-terminal region regulates the binding of PLA 1 to PC. A hypothesis is presented according to which the α9 helix of C-terminal region in PLA 1 not only controls the opening of lid but also serves as a membrane anchor that assists in binding to PC. These findings bring new insight into rational design of novel lipases with intriguing functionalities.
研究领域:
酶学
·
生物化学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
LWT-FOOD SCIENCE AND TECHNOLOGY
·
2024
·
IF 6
Lilang Li,Kuntai Li,Ruiguo Cui,Fengjun Li,Jianrong Wang
Lipase from Thermomyces dupontii (TDL) shows great potential to prepare long-medium-long structured lipids (LML-SLs), but its poor thermostability represents a significant obstacle to further industrial application. Herein, a rational design based on structure and sequence analysis (including disulfide bond calculation and sequence alignment) was used to generate a more thermostable mutant. Compared with wild-type TDL, mutant M3 displayed a 9.1-fold, 7.2 °C and 10 °C increase in half-life ( t 1/2 ) at 60 °C, the half-loss temperature ( T 50 ) at 15 min and optimum temperature, respectively. Remarkably, the specific activity of mutant M3 was 49.1% higher than that of wild-type TDL. Molecular dynamics (MD) simulations showed that the structural flexibility of mutant M3 was reduced, which might contribute to the enhanced thermostability. Additionally, LML-SLs were synthesized from ethyl linoleate and tricaprylin with wild-type TDL and mutant M3. Compared with the wild-type TDL group, the LML-SL content of the mutant M3 group increased by 8.1%. This study will pave the way for the industrial application of TDL and provide valuable insights for enhancing the thermostability of other lipases.
研究领域:
生物化学
·
酶工程
·
食品科学
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产品关联
DOI
高影响因子
Processes
·
2020
·Vol. 8
·期 5
·629
·
IF 2.753
Yanhong Bi,Zhaoyu Wang,Rui Zhang,Yihan Diao,Yaoqi Tian,Zhengyu Jin
In this study, magnetic Fe3O4nanoparticles coated with polydopamine possessing abundant amino groups (Fe3O4@PDA) were conveniently prepared, detailed, and characterized, and then firstly used as a supporting matrix for immobilizingThermomyces lanuginosuslipase (Fe3O4@PDA@TLL). The effects of some crucial factors on the immobilization efficiency were investigated and the optimal protein loading and activity recovery were found to be 156.4 mg/g and 90.9%, respectively. Characterization studies revealed that Fe3O4@PDA@TLL displayed a broader pH and temperature adaptability as compared to the free TLL, which allows its use at wider ranges of reaction conditions. With regard to the stabilities, the immobilized TLL clearly displayed improved pH, thermal, and solvent tolerance stabilities compared to the free enzyme, suggesting that the biocompatible Fe3O4@PDA might be an outstanding material for immobilizing TLL and acting as alternative support for different enzymes.Keywords:magnetic nanoparticle;immobilization;Thermomyces lanuginosuslipase;stability
研究领域:
生物化学
·
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）