CHEMICAL ENGINEERING SCIENCE
·
2019
·
IF 3.372
Huan Xia,Zhixian Li,Xue Zhong,Binghui Li,Yunpeng Jiang,Yanbin Jiang
The combination of enzymatic and chemical reactions to achieve co-operative chemo-enzymatic processes has presented many challenges for decades. In this work, alcohol dehydrogenase (ADH) was encapsulated into metal-organic framework HKUST-1 crystals through an in situ approach to construct a synergistic chemo- and bio-catalysis system, where ADH was for the conversion of benzyl alcohol to benzaldehyde, and HKUST-1 was in charge for the regeneration of β-nicotinamide adenine dinucleotide (NAD + , the oxidized cofactor). It was found that HKUST-1 was a versatile catalyst for efficient oxidation of NADH in different buffers. The Michaelis-Menten kinetic parameters of immobilized ADH exhibited an increased K M (approximately 1.3-fold) and a decreased V max (approximately 4.7-fold) compared to free ADH. Due to the protection of HKUST-1, the immobilized enzyme showed a better resistance against urea and organic solvents compared with free enzyme, and still maintained 93% of its original activity after 6 cycles. The immobilized enzyme was successfully applied to the enzymatic transformation of benzyl alcohol to benzaldehyde in aqueous media at room temperature. Owing to the oxidized cofactor regeneration by HKUST-1, ADH@HKUST-1 composites achieved a benzaldehyde yield of 28.5%, which was approximately 4.3-fold higher than that of the individual bio-catalysis. This concept of combining chemo- and bio-catalysis may provide a novel and versatile technique for applications in biochemical catalysis, biosensors and drug delivery.
研究领域:
生物化学
·
催化科学
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产品关联
DOI
高影响因子
Catalysis Science & Technology
·
2024
·
IF 4.4
Jianhua Liu,Jiashu Li,Fangshu Xing,Jian Liu
Developing biological cofactor regeneration strategies is crucial for photo-enzyme coupling systems. Here, crystalline poly(triazine imide) (PTI) containing the cyano group (–CN) is designed for photocatalytic nicotinamide cofactor oxidation regeneration via π–π interaction. The π-conjugation degree and electron delocalization of PTI can be regulated through –CN modification. Such a unique extended π-conjugated system promotes the adsorption of the reduced-state nicotinamide cofactor (NADH) on the aromatic skeleton. Moreover, the enhanced charge transfer dynamics expedite the formation of intermediate active species. Accordingly, the optimized conversion rate of NADH to NAD+ reached 98.2% within 60 min. In contrast, pristine PTI and amorphous polymeric carbon nitride (PCN) gave oxidation yields of 88.1% and 56.8%, respectively. The exclusive selectivity of the oxidized product was further verified by the enzymatic method and nuclear magnetic resonance characterization. This study introduces an innovative method for modulating non-covalent interactions for photo-enzymatic catalysis via the extended π-conjugated system.
研究领域:
材料科学
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光化学
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生物催化
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产品关联
DOI
高影响因子
GREEN CHEMISTRY
·
2024
·
IF 9.3
Si-Yu Zhu,Shi-Chang Liu,Chuan-Xi Zhang,Xin Xin,Zhi-Hua Liu,Lujia Zhang,Bing-Zhi Li,Ying-Jin Yuan
A sustainable lignin bioconversion depends on the exploitation of the inherent aromaticity of lignin for high-value aromatic products. However, the microbial valorization of lignin is hindered by the lack of established bioconversion routes or the inefficient heterologous expression of key enzymes. Here, an efficient biological lignin conversion route was constructed in a microbial cell factory by tapping inherent aromaticity of lignin to produce aromatic natural compound such as homoeriodictyol. The heterologous biosynthesis pathways were successfully constructed to convert lignin-derived monomers to homoeriodictyol in Saccharomyces cerevisiae. The implementation of combinatorial regulating strategies resulted in enhanced CoA supply, suppression of branch pathways, and upregulation of the rate-liming enzymes, which increased the titer of homoeriodictyol by a remarkable 22.5-fold. After that, engineering cofactor supply and recycling led to a substantial 95% increase in homoeriodictyol production. The rational design and modification of the heterologous rate-limiting enzymes achieved substantial improvement in the homoeriodictyol yield. All these strategies together enabled the record homoeriodictyol titer of 3.2 mmol/l from lignin derivatives by S. cerevisiae. Overall, such innovative conversion route meets the atom-economic concept by converting lignin derivatives into valuable flavonoid, paving a green, sustainable way for both the valorization of lignin and the production of aromatic natural products.
研究领域:
生物技术
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代谢工程
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生物化学
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微生物学
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DOI
高影响因子
Bioresource Technology Reports
·
2022
·
IF 0
Jiali Mi,Simin Liu,Yan Du,Haishan Qi,Lei Zhang
Cadaverine (1,5-diaminopentane) has widespread applications in industry, agriculture and medicine. Cadaverine can be biotransformed from l -lysine by lysine decarboxylasey which is a pyridoxal 5′-phosphate (PLP)-dependent enzyme. Herein, barium alginate microspheres were fabricated by electro-dripping and employed as immobilized metal ion affinity chromatography matrix to purify and immobilize engineered lysine decarboxylase CadA-EK. Besides, polyethyleneimine (PEI) was employed for co-immobilizing PLP and enzymes (cCadA-EK-PEI/PLP). The cCadA-EK-PEI/PLP can produced 184.53 g/L cadaverine with yield of 90.03% in 2 mol/L lysine solution. And the stability of co-immobilized CadA-EK was 1–4 times higher than that of iCadA-EK + PLP in environmental factors, and the catalytic efficiency k cat / K m was increased by 3.4-fold. Furthermore, a continuous flow microreactor with the co-immobilized enzymes and cofactors was developed to catalyze lysine, with 14.28 g/(L·h) productivity of cadaverine, which displayed great potential for cadaverine biosynthesis. This work provides alternative strategy for the one-step purification and co-immobilization of enzyme and cofactor.
研究领域:
生物催化
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酶工程
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生物合成
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材料科学
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DOI
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
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2025
·Vol. 26
·期 10
·4795
·
IF 4.9
Tingxiao Guo,Lifang Zeng,Jiaxu Liu,Xiaoyan Zhang,Yunpeng Bai
Enzyme catalysis represents a promising approach for sustainable chemical synthesis, yet its industrial applications face limitations due to the inefficient regeneration and high cost of essential cofactors, such as adenosine-5′-triphosphate (ATP) and nicotinamide adenine dinucleotide phosphate (NADPH). While natural metabolic systems efficiently recycle cofactors through spatially organized enzymes, replicating this efficiency in vitro remains challenging. Here, we prepare a five-enzyme condensate system using liquid–liquid phase separation (LLPS) mediated by intrinsically disordered proteins (IDPs). By colocalizing a carboxylic acid reductase from Norcadia iowensis ( Ni CAR) with a reductive aminase from Aspergillus oryzae ( Asp RedAm) and three cofactor-regenerating enzymes, we generated a phase-separated catalytic condensate that enhanced ATP and NADPH recycling efficiency by 4.7-fold and 1.9-fold relative to free enzymes, respectively. Catalytic performance was correlated with the extent of phase separation, as confirmed by fluorescence microscopy, which revealed clear enrichment of ATP and NADPH within the condensates. This proximity effect enabled efficient cofactor turnover in the one-step reaction, achieving substrate conversion above 90% within 6 h and enhancing the space–time yield (STY) of the chiral imines 1.6-fold, with only one-fifth of the standard cofactor load. This approach creates a scalable and economic tool for performing multienzyme cascade reactions in vitro that are driven by the efficient recycling of multiple cofactors.
研究领域:
生物化学
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合成生物学
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DOI
PubMed
高影响因子
Molecular Catalysis
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2022
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IF 5.089
Weiwen Wang,Jianing Qu,Chaojie Li,Lexin Guo,Xianxin Fang,Guanghui Chen,Jihai Duan
Biomimetic photocatalytic nitrogen fixation is considered as a potential alternative to the Haber-Bosch method, which is an ideal pathway for nitrogen reduction. The development of efficient bionanocatalysts is a serious challenge in the current field of biomimetic nitrogen fixation. Here, we introduced a second metal in the MIL-88A/Fe mesh by metal doping to build a Mo-doped MIL-88A(Fe/Mo)/Fe mesh biomimetic system. The catalyst is a rod-like structure synthesized in situ by hydrothermal method using iron mesh as the substrate, and its rough surface is favorable for the adsorption of N 2 . The introduction of Mo provides a two-electron transfer pathway for the reaction, and the formation of the "MoFe cofactor" effectively promotes charge separation and inhibits the photoelectron-hole complexation. In addition, we kept the catalyst above the liquid surface and provided the proton source through water vapor, which solved the problem of the low solubility of N 2 in water and poor mass transfer efficiency. The results show that the biomimetic photocatalyst has good photoresponsiveness in the whole visible region, and the synergistic effect between the two metals makes the MIL-88A (Fe/Mo 0.017 )/Fe mesh exhibit better photocatalytic performance, and its nitrogen fixation efficiency reaches about 9.6 times that of the original MIL-88A/Fe mesh. This study provides a new idea for the design of photocatalytic ammonia biomimetic photocatalysts.
研究领域:
材料科学
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环境科学
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生物化学
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DOI
高影响因子
JOURNAL OF MOLECULAR LIQUIDS
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2020
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IF 5.065
Xiangqian Li,Qin Wen,Jiapei Gu,Wanqiang Liu,Qianming Wang,Guofu Zhou,Jinwei Gao,Yuhui Zheng
A new Schiff base receptor (2-amino-3-(((3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl)methylene)amino)maleonitrile ( GAL )) has been synthesized and such diaminomaleonitrile-based molecular framework is observed to be water soluble . GAL possesses both colorimetric and “off-on” fluorescent response in the presence of ClO − . The response time has been controlled within 6 min. The limit of detection (LOD) has been calculated to be 47.5 nM. The addition of Cu 2+ can only induce clear color evolution from pale to deep yellow (LOD: 0.22 μM) and no fluorescence changes are found. Moreover, its reliability and practicality are verified via the determination of ClO − in spiked samples of tap water and pond water. The exploration of bioactive vitamin B6 cofactor as a sensing platform will open a new way for multiple target recognition in competitive mediums.
研究领域:
化学
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生物化学
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DOI
高影响因子
ACS Sustainable Chemistry & Engineering
·
2024
·
IF 7.1
Si Liu,Xiaohui Wu,Xiaoyan Dong,Qinghong Shi,Yan Sun
Large-scale applications of enzymes and cofactors remain a challenge due to their fragile property, nonrecyclability, and high cost. Herein, we report a coimmobilization strategy of multienzyme and cofactor based on a combination of microgel and silica nanoparticles (SNPs) to make the biocatalysis process economical and sustainable. As exemplified by a nicotinamide adenine dinucleotide (NAD)-dependent alcohol dehydrogenase and chimeric amine dehydrogenase cascade amination reaction, the dual-enzyme coimmobilization system, with 100% immobilization yield and 76% increased cascade activity, was constructed through the site-specific immobilization of silica binding peptide. The anionic dual-enzyme@SNPs (29.7 mg gSNPs/PVCL-Vim–1) and NADH (47.3 μmol gSNPs/PVCL-Vim–1) are then delivered into a cationic microgel through gel swelling and electrostatic interactions/ion exchanges. The system displays a 27 to 75% enhancement in the amination yield as compared to the free counterpart, possibly due to the site-specific immobilization of enzymes, the proximity effect resulting from the coimmobilization, and the mobility and accessibility of the immobilized cofactor. Moreover, the constructed coimmobilization system shows 8–43% increased stability over the free system and retains 97% of its original activity after eight recycles. Collectively, this work has demonstrated an efficient coimmobilization strategy of a cofactor-dependent multienzyme cascade for applications of sustainable biotransformation.
研究领域:
生物化学
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材料科学
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酶工程
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产品关联
DOI
高影响因子
CHINESE JOURNAL OF CATALYSIS
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2025
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IF 17.7
Jiaxu Liu,Jiaxin Chen,Xiaoyan Zhang,Daidi Fan,Yunpeng Bai
Enzyme catalysis is a promising way to produce chiral products in a green and sustainable way. However, the high cost of cofactors and their relatively low recycling efficiency hinder the widespread application of enzyme catalysis in industry. In contrast, cofactor regeneration and recycling in cells is highly efficient, mainly due to physical effects caused by the ordered spatial organization of enzymes in vivo . The construction of similar catalytic systems with high cofactor recycling in vitro remains challenging. Here, we present a facile method to generate dual enzyme condensates in vitro based on intrinsically disordered region-mediated liquid-liquid phase separation. Typically, a carbonyl reductase from Serratia marcescens ( Sm CR V4 ) and a glucose dehydrogenase from Bacillus megaterium ( Bm GDH) were co-localized in the condensates. This resulted in an up to 20-fold increase in cofactor recycling efficiency (substrate converted per cofactor per unit time), and a 3.4-fold increase in space-time yield compared to the free enzyme system. The reaction enhancement was shown to be highly correlated with the degree of condensation of the dual enzymes. Fluorescence confocal microscopy showed that the cofactor, nicotinamide adenine dinucleotide phosphate (NADPH), was enriched between neighboring enzymes during the reaction due to the proximity effect, facilitating its regeneration and recycling within the condensate. In a scaled-up synthesis, the consumption of NADPH was reduced 50-fold compared to industrial biocatalytic standards, while the condensate still maintained efficient product synthesis. Concentrating multiple enzymes in a nano- and micro-condensate to increase the reaction rate may provide a general and inexpensive method for improving cofactor-involved enzymatic reactions.
研究领域:
生物技术
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酶学
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化学工程
✓ 已验证
DOI
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
·
2023
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IF 3.4
Lu Zhao,Wenhe Zhang,Qi Wang,Huibin Wang,Xiao Gao,Bin Qin,Xian Jia,Song You
L -Phosphinothricin ( L -PPT) is the effective constituent in racemic PPT (a high-efficiency and broad-spectrum herbicide), and the exploitation of green and sustainable synthesis route for L -PPT has always been the focus in pesticide industry. In recent years, “one-pot, two-step” enzyme-mediated cascade strategy is a mainstream pathway to obtain L -PPT. Herein, Rg DAAO and Bs LeuDH were applied to expand “one-pot, two-step” process. Notably, a NADH-dependent leucine dehydrogenase from Bacillus subtilis ( Bs LeuDH) was firstly characterized and attempted to generate L -PPT, achieving an excellent enantioselectivity (99.9% ee ). Meanwhile, a formate dehydrogenase from Pichia pastoris ( Pp FDH) was utilized to implement NADH cofactor regeneration and only CO 2 was by-product. Sufficient amount of the corresponding keto acid precursor PPO was obtained by oxidation of D -PPT relying on a D -amino acid oxidase from Rhodotorula gracilis ( Rg DAAO) with content conversion (46.1%). L -PPT was ultimately prepared from racemized PPT via oxidative deamination catalyzed by Rg DAAO and reductive amination catalyzed by Bs LeuDH, achieving 80.3% overall yield and > 99.9% ee value.
研究领域:
生物化学
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农药合成
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酶工程
✓ 已验证
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2023
·
IF 6.1
Lian Wang,Huijing Wang,Jianbin Chen,Minglong Hu,Xiaoyu Shan,Jingwen Zhou
Chlorogenic acid is a natural phenolic compound widely used in the food and daily chemical industries. Compared to plant extraction, microbial cell factories provide a green and sustainable production method for the production of chlorogenic acid. However, complex metabolic flux distribution and potential byproducts limited the biosynthesis of chlorogenic acid in microorganisms. A de novo biosynthesis pathway for chlorogenic acid was constructed in Escherichia coli via modular engineering. Increasing the shikimate pathway flux greatly promoted chlorogenic acid production, and the influence of pyruvate metabolism on chlorogenic acid synthesis was also explored. The supply of cofactors for the key enzymes quinate/shikimate 5-dehydrogenase (YdiB) and 4-hydroxyphenylacetate 3-monooxygenase (HpaBC) was enhanced by a cofactor regeneration system. Furthermore, mutants of YdiB were verified for chlorogenic acid production in vivo. Chlorogenic acid browning occurred when the buffer pH of the buffer exceeded 6.0, but two-stage pH control achieved a chlorogenic acid titer of 2789.2 mg/L in a 5 L fermenter, the highest reported to date. This study provided a strategy for the efficient production of chlorogenic acid from simple carbon sources.
研究领域:
生物工程
·
微生物学
·
合成生物学
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产品关联
DOI
PubMed
高影响因子
Microorganisms
·
2023
·Vol. 12
·期 1
·38
·
IF 4.5
Kun Geng,Ying Lin,Xueyun Zheng,Cheng Li,Shuting Chen,He Ling,Jun Yang,Xiangyu Zhu,Shuli Liang
Acetaldehyde is an important carbonyl compound commonly detected in wines. A high concentration of acetaldehyde can affect the flavor of wines and result in adverse effects on human health. Alcohol dehydrogenase I (ADH1) inSaccharomyces cerevisiaecatalyzes the reduction reaction of acetaldehyde into ethanol in the presence of cofactors, showing the potential to reduce the content of acetaldehyde in wines. In this study,ADH1was successfully expressed inPichia pastorisGS115 based on codon optimization. Then, the expression level ofADH1was enhanced by replacing its promoter with optimized promoters and increasing the copy number of the expression cassette, with ADH1 being purified using nickel column affinity chromatography. The enzymatic activity of purified ADH1 reached 605.44 ± 44.30 U/mg. The results of the effect of ADH1 on the content of acetaldehyde in wine revealed that the acetaldehyde content of wine samples was reduced from 168.05 ± 0.55 to 113.17 ± 6.08 mg/L with the addition of 5 mM NADH and the catalysis of ADH1, and from 135.53 ± 4.08 to 52.89 ± 2.20 mg/L through cofactor regeneration. Our study provides a novel approach to reducing the content of acetaldehyde in wines through enzymatic catalysis.Keywords:alcohol dehydrogenase I;Pichia pastoris;cofactor regeneration;multicopy;acetaldehyde;wine
研究领域:
微生物学
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生物技术
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食品科学
✓ 已验证
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）