iScience
·
2024
·
IF 4.6
Jiawei Wang,Xingyu Ouyang,Shiyu Meng,Jiayi Li,Liangxu Liu,Chaofeng Li,Hengrun Li,Haotian Zheng,Chao Liao,Yi-Lei Zhao,Jun Ni
Lignin valorization is crucial for achieving economic and sustainable biorefinery processes. However, the enzyme substrate preferences involved in lignin degradation remain poorly understood, and low activity toward specific substrates presents a significant challenge to the efficient utilization of lignin. In this study, we investigated the substrate promiscuity of Th Ado, a key enzyme involved in lignin valorization. Pre-reaction state analysis revealed that a hydrogen bond network is critical in determining substrate selectivity. By performing targeted saturation mutagenesis on residues surrounding the substrate tunnels, we identified the Y205W and Y205Q mutants, which demonstrated 0.73-fold and 0.72-fold enhancements in activity, respectively. Structural analysis indicated that the redirection of the original substrate tunnel may be responsible for the improved activity. Our study provides essential insights into the substrate preference mechanisms of lignin degrading enzymes and suggests that this tunnel-redirection strategy can be extended to other promiscuous enzymes.
研究领域:
生物化学
·
生物技术
·
酶学
·
可持续化学
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产品关联
DOI
PubMed
高影响因子
APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY
·
2022
·Vol. 194
·期 11
·5119-5131
·
IF 3.094
Yang Yang,Wang Jia,Yang Bo,Lan Dongming,Wang Yonghua
The amino acid residues lining the substrate binding pocket play quite an important role during the lipase catalytic process. The conversion of those residues might cause a dramatic change in the lipase properties, such as the substrate selectivity of lipase. In our study, T237 residue sitting on the entrance of the catalytic pocket in lipase MAS1 was important for the catalytic performance. When replacing polar Thr with the positively charged Arg, the synthesis ratio of partial glycerides/triglycerides increases to 6.32 rather than 1.21 of MAS1 wild type (WT), as the substrate ratio of glycerol and fatty acids is 1:3. And the fatty acid preference shifted to long-chain substrates for mutant T237R rather than middle-chain substrates for MAS1 WT. Molecular docking analysis revealed that hydrophobic and side chain properties of Arg might contribute to the change of the MAS1 lipase catalytic performance. This work would pave a way for the accurate rational transformation of the lipases to produce value lipid for industrial application. Graphical abstract
研究领域:
生物化学
·
酶学
·
生物技术
·
蛋白质工程
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产品关联
DOI
PubMed
高影响因子
CABI Agriculture & Bioscience
·
2023
·Vol. 4
·期 1
·1-8
·
IF 3.9
Huang Hexi,Di Ning,Wang Jie,Wang Yuxing,Zhu Zhengyang,Lu Caige,Wang Su,Zang Liansheng
The heavy metal, cadmium (Cd), causing growth retardation and yield reduction on rice and impacting the fitness of organisms inhabiting on rice through bottom-up effects, has become a great challenge to rice production. However, the effect of Cd-exposure on the development of an economically important and destructive rice pest, Chilo suppressalis remains unexplored. By exposing the larvae of C. suppressalis to different Cd-exposed artificial diets (0, 0.2, 1.0, 2.5, 5.0, and 10.0 mg/kg), we found that Cd exposure did not affect the larval duration or pupation rate of C. suppressalis , but caused negative effects on pupal weight at high Cd levels (5.0 and 10.0 mg/kg) and on adult deformity rate from 2.5 and 5.0 mg/kg treatments. Although Cd significantly increased the female pupae ratio, C. suppressalis did not oviposit when Cd treatment was more than 2.5 mg/kg. Meanwhile, Cd transferred to pupae, females, exuviae of pupa and eggs of C. suppressalis from Cd treated larvae, and exhibited a dose-dependent response on Cd accumulation. Our results indicated that Cd had a negative effect on rice stem borer and can be transferred to eggs of C. suppressalis , but more work is needed to further assess the bottom-up effect on third tropic levels in Cd-polluted fields.
研究领域:
生态毒理学
·
昆虫学
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产品关联
DOI
高影响因子
SENSORS
·
2016
·Vol. 16
·期 4
·584
·
IF 2.033
Jiaheng Sun,Chunyan Li,Yanfei Qi,Shuanli Guo,Xue Liang
Nanozyme-based chemical sensing is a rapidly emerging field of research. Herein, a simple colorimetric assay for the detection of hydrogen peroxide and glucose based on the peroxidase-like activity of V2O5nanozymes has been established. In this assay, the effects of pH, substrate, nanozyme concentrations and buffer solution have been investigated. It was found that compared with 3,3′,5,5′-tetramethylbenzidine (TMB), the enzyme substrateo-phenylenediamine (OPD) seriously interfered with the H2O2detection. Under the optimal reaction conditions, the resulting sensor displayed a good response to H2O2with a linear range of 1 to 500 μM, and a detection limit of 1 μM at a signal-to-noise ratio of 3. A linear correlation was established between absorbance intensity and concentration of glucose from 10 to 2000 μM, with a detection limit of 10 μM. The current work presents a simple, cheap, more convenient, sensitive, and easy handling colorimetric assay.Keywords:V2O5nanozymes;variation;substrates;H2O2;glucose
研究领域:
生物化学
·
医学检测
·
纳米技术
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产品关联
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2021
·
IF 3.493
Xinqi Xu,Xiangzhen Deng,Juan Lin,Jie Yang
Arylsulfatase is useful in industrial agar processing by removing sulfate groups. A full-length arylsulfatase gene, designated ArySMA1, was obtained from marine bacteria Serratia sp. SM1. The ArySMA1 gene encoded a novel serine-type arylsulfatase and the enzymatic properties were characterized. The enzyme presented notable capacity of removing sulfate groups from natural algae substrates. Kinetic study suggested that the microscopic thermal inactivation rate of ArySMA1 in free form was smaller than that of the enzyme-substrate complex. The presence of substrate could unexpectedly accelerate ArySMA1 to deactivate at high temperature. Such phenomenon was opposite to many findings that substrate could stabilize enzymes against heat. Molecular dynamics simulation and ANS fluorescent assay indicated the substrate led the hydrophobic regions of the active site more flexible and the sulfate group of the substrate could retard the processivity of ArySMA1 catalysis. This study provides guidance for agar desulfation and down-stream processing industry.
研究领域:
酶学
·
海洋生物技术
·
工业生物技术
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产品关联
DOI
PubMed
高影响因子
Results in Engineering
·
2025
·
IF 7.9
Wei Kuang,Yuxiang Hong,Zimu Hu,Peng Xu
Liquid C1 feedstock, including methanol, formaldehyde and formate can be easily manufactured from greenhouse gases CO 2 and CH 4 . Yarrowia lipolytica , isolated from marine ecosystem, is a non-conventional yeast which is capable of assimilating multiple range of hydrophobic compounds including alcohols, lipids, hydrocarbons, and volatile fatty acids. This yeast has been extensively engineered as a benchmark host to produce lipids, resveratrol, carotenoids, squalene and other natural products. Connecting C1 feedstock pathways to these important metabolites holds promise to further reduce costs and improve sustainability. Herein we tested the growth fitness of Y. lipolytica when exposed to various one-carbon (C1) liquid feedstocks (methanol, formaldehyde, and formate). We firstly examined the cell growth kinetics with an automatic Cell Growth Analyzer and fitted the data with a Monod-type growth model. We obtained the inhibition constants of the three liquid C1s from the growth fitness dose-response curves. We found that the lagging phase duration is exponentially correlated with the toxicity of the tested C1 solvents. Notably, Y. lipolytica exhibited tolerance to methanol (IC 50 871 mM) and formate (IC 50 42.6 mM), but showed sensitivity to formaldehyde (IC 50 3.8 mM). We further demonstrated that glucose and formate co-substrate could improve cell growth and lipid production. This work broadens Y. lipolytica ’s substrate scope and facilitates sustainable and cost-efficient production of fuels and chemicals from C1 feedstocks.
研究领域:
微生物学
·
生物工程
·
环境科学
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产品关联
DOI
高影响因子
EUROPEAN FOOD RESEARCH AND TECHNOLOGY
·
2014
·Vol. 240
·期 3
·627-634
·
IF 1.387
Ma Huihui,Zhang Niya,Sun Lvhui,Qi Desheng
Aflatoxin contamination levels vary from crop to crop, suggesting that it is of great significance to explore the effects of different substrates on aflatoxins production. In this study, the effects of corn, wheat, peanut, and soybean on aflatoxin B 1 (AFB 1 ) production by Aspergillus parasiticus were investigated, and the effects of corn germ, wheat germ, soybean, and peanut oils on the production of AFB 1 by A . parasiticus in the corresponding defatted substrates were evaluated at the rates of 2, 5, 10, and 15 g/100 g of substrate. The effects of the combinations of different oils with different defatted substrates on the accumulation of AFB 1 by A . parasiticus were also studied. AFB 1 concentration was determined by HPLC. The results indicated that high-lipid substrates of soybean and peanut had significantly higher AFB 1 production than low-fat substrates of wheat and corn throughout the entire incubation period (15 days) in both autoclaved and non-autoclaved grains. Corn germ, wheat germ, soybean, and peanut oils with different doses could effectively stimulate AFB 1 accumulation, and AFB 1 production was determined by the oil content of the substrates. Our results suggest that the oil content of substrates is one of the factors affecting AFB 1 production by A . parasiticus and that the composition and content of the substrates may also play a role.
研究领域:
食品科学
·
微生物学
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产品关联
DOI
高影响因子
CHEMICAL ENGINEERING JOURNAL
·
2023
·
IF 15.1
Zhaoxin Wang,Jinlan Wen,Jihang Zhang,Jiawei Deng,Wei Zhuang,Jinle Liu,Zhi Wang,Yuan Rao,Yudan Zhu,Hanjie Ying
Lipase catalysis plays an important role in bioengineering and food science and is widely used in the organic solvent environment. The addition of trace water to organic media can significantly change lipase activity, and the reason is essential to understand. In this study, by combining experiments and molecular dynamics simulations, we studied solvent-free acidolysis of the tripalmitin (PPP) with different water contents and revealed the mechanism of the water effect on lipase activity from atomic insight. Enzyme activity experimentally exhibited an increasing but subsequently decreasing trend with the rise of water content. Through molecular dynamics (MD) simulations, we found that the reaction activity was affected by three aspects: lipase flexibility, substrate diffusion and substrate spatial distribution. Lipase flexibility, as the structural basis of catalytic activity, increased with water content. At lower water content, the diffusion ability of the substrate around the lipase was dominant. With water content increasing, water molecules on the protein surface gradually shielded hydrogen bonds (HBs) between the oleic acids (OA) and the lipase, which improved the diffusion ability of the substrate near the enzyme surface and thus beneficial to the reaction occurring. At higher water content, the spatial distribution dominated the reaction, where the thicker water layer on the lipase surface acted as a spatial barrier hindering the access of substrate OA to the enzyme and thus harmful to the reaction. Our work helps to provide guidance on controlling enzyme activities in organic solvent systems.
研究领域:
生物工程
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食品科学
·
酶学
·
分子动力学
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产品关联
DOI
高影响因子
MOLECULAR BIOTECHNOLOGY
·
2016
·Vol. 58
·期 6
·428-440
·
IF 1.752
Yang Yuanyuan,Lin Jinping,Wei Dongzhi
A NADPH-dependent and FMN-containing nitroreductase (Gox0834) from Gluconobacter oxydans was cloned and heterogeneously expressed in Escherichia coli . The purified enzyme existed as a dimer with an apparent molecular mass of about 31.4 kDa. The enzyme displayed broad substrate specificity and reduced a variety of mononitrated, polynitrated, and polycyclic nitroaromatic compounds to the corresponding amino products. The highest activity was observed for the reduction of CB1954 (5-(1-aziridinyl)-2,4-dinitrobenzamide). The enzyme kinetics analysis showed that Gox0834 had relatively low K
m
(54 ± 11 μM) but high k
cat / K
m
value (0.020 s −1 /μM) for CB1954 when compared with known nitroreductases. Nitrobenzene and 2,4,6-trinitrotoluene (TNT) were preferred substrates for this enzyme with specific activity of 11.0 and 8.9 μmol/min/mg, respectively. Gox0834 exhibited a broad temperature optimum of 40–60 °C for the reduction of CB1954 with a pH optimum between 7.5 and 8.5. The purified enzyme was very stable below 37 °C over a broad pH range of 6.0–10.0. These characteristics suggest that the nitroreductase Gox0834 may be a possible candidate for catalyzing prodrug activation, bioremediation, or biocatalytic processes.
研究领域:
微生物学
·
酶学
·
环境科学
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产品关联
DOI
PubMed
高影响因子
CATALYSIS COMMUNICATIONS
·
2013
·
IF 2.915
Hong-De Yan,Qun Zhang,Zhao Wang
A lipase from Aspergillus oryzae WZ007 exhibited high esterification activities toward a series of short-chain acids and alcohols. Moreover, it displayed a strong tolerance for high substrate concentration of up to 2.0 M and presented a highest initial rate of 276 mmol·L − 1 ·h − 1 at this concentration. After a reaction time of 48 h, the conversion rates of acids were higher than 80% for the synthesis of a majority of heptanoic acid esters, some octanoic acid esters and n -propyl hexanoate. These results implied that A. oryzae WZ007 lipase was a promising biocatalyst in the production of flavor esters.
研究领域:
生物化学
·
食品科学
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产品关联
DOI
高影响因子
Foods
·
2022
·Vol. 11
·期 7
·980
·
IF 5.561
Jiamei Xu,Yu Liu,Jingjing Ma,Pengpeng Li,Zhiming Geng,Daoying Wang,Muhan Zhang,Weimin Xu
Lipoxygenase (LOX)is a major endogenous enzyme for the enzymatic oxidation of lipids during meat storage and meat product manufacturing. In the present work, some characteristics, i.e., effects of inhibitors, selectivity of substrates and specificity of oxidation products, were studied using recombinant porcine 12-lipoxygenase catalytic domain(12-LOXcd). Several familiar inhibitors were found inhibit the activity of recombinant porcine 12-LOXcd;nordihydroguaiaretic acid demonstrated the strongest inhibitory effect. The enzyme could oxygenate common polyunsaturated fatty acids, and showed the highest affinity to linoleic acid (LA), followed by arachidonic acid (AA), linolenic acid (LN) and docosahexaenoic acid (DHA). Under the action of porcine 12-LOXcd, LA was oxidized into four hydroxyoctadecadienoic acid (HODE) isomers, i.e., 13-Z,E-HODE, 13-E,E-HODE, 9-Z,E-HODE and 9-E,E-HODE. Variation of pH not only affected the yield of LA oxidation products, but also the distribution of HODE isomers. These results indicated that endogenous LOX activity and LOX-catalyzed lipid oxidation can be regulated during meat storage and meat product manufacturing.Keywords:Lipoxygenase;inhibitors;substrates;oxidation products;linoleic acid;porcine
研究领域:
食品科学
·
生物化学
·
酶学
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产品关联
DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
·
2022
·
IF 11.889
Xiaoyu Chen,Zhe Dou,Tianwei Luo,Zewen Sun,Hongmin Ma,Guochao Xu,Ye Ni
Ancestral enzymes are promising for industrial biotechnology due to high stability and catalytic promiscuity. An effective protocol was developed for the directed resurrection of ancestral enzymes. Employing genome mining with diaryl alcohol dehydrogenase Kp ADH as the probe, descendant enzymes D10 and D11 were firstly identified. Then through ancestral sequence reconstruction, A64 was resurrected with a specific activity of 4.3 U·mg −1 . The optimum pH of A64 was 7.5, distinct from 5.5 of D10. The T15 50 and T m values of A64 were 57.5 °C and 61.7 °C, significantly higher than those of the descendant counterpart. Substrate spectrum of A64 was quantitively characterized with a Shannon-Wiener index of 2.38, more expanded than D10, especially, towards bulky ketones in Group A and B. A64 also exhibited higher enantioselectivity. This study provides an effective protocol for constructing of ancestral enzymes and an efficient ancestral enzyme of industrial relevance for asymmetric synthesis of chiral alcohols.
研究领域:
生物化学
·
工业生物技术
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DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）