JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Zhuangzhuang Huang,Junru Zhou,Jialing Wang,Sheng Xu,Cheng Cheng,Jiangfeng Ma,Zhen Gao
The industrial applications of enzymes are limited due to the activity-stability trade-off, which implies that the improvement of thermostability often accompanies decreased activity. This study presents a dual-strategy approach to simultaneously improve the catalytic efficiency and thermostability of α-galactosidase galV from Anoxybacillus vitaminiphilus WMF1. Our integrated method combines computational analysis with enzyme property prediction to selectively target and modify the catalytic region and residues that are distant from the active site. We identified and experimentally validated mutations that improve activity without compromising stability and further increased thermostability through additional distant-site mutations. The resulting mutant enzyme variant N549Q/T550N/Y634F demonstrated a 6.2-fold increase in catalytic efficiency and a 3.2-fold improvement in the half-life at 65 °C. Molecular dynamics (MD) simulations supported the structural basis for the observed enhancements. This approach offers a refined strategy for engineering α-galactosidases with improved industrial applicability, overcoming the traditional trade-offs between enzyme activity and stability. Hydrolytic activity toward raffinose family oligosaccharides (RFOs) was validated using soymilk as a model substrate, demonstrating significant practical potential.
研究领域:
生物化学
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酶工程
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计算生物学
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DOI
PubMed
高影响因子
Food Bioscience
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2024
·
IF 4.8
Jingjing Dong,Yanli Wang,Xiaoxiao Li,Ying Chen,Rui Fan,Nana Wang,Zhengyu Jin,Yuxiang Bai
The starch-converting GH70 α-glucanotransferases, as the influential enzymatic tools for the synthesis of structurally diverse α-glucans, are of great value in the food industry. In this work, a new GtfB-type α-glucanotransferase from Fructilactobacillus sanfranciscensis Gs2 (FsGs2 GtfB) was mined from the NCBI database, which is highly conserved in four conserved regions I-IV for GtfB enzymes but possessed distinct characteristics in some residues at acceptor subsites +1/+2. Biochemical characterizations revealed that the FsGs2 GtfB enzyme demonstrated a novel 4,6-α-glucanotransferase activity, which was able to cleave the (α1 → 4) linkages in maltooligosaccharide/starch substrates and mainly synthesize branching (α1 → 4,6) linkages in the gelatinized system, resulting in the generation of low-molecular-mass short-clustered (α1 → 4,6)-glucans. Nevertheless, FsGs2 GtfB acted as a quasi-hydrolase instead of transglucosylase in the granular system, causing a notable decrease in the molecular mass of granular wheat starch and a tendency towards shortened chain lengths. Hence, the FsGs2 GtfB enzyme represents an additional potent enzymatic tool of reassembling the (α1 → 4) and (α1 → 6) linkages in starch molecules and enriches the diversity of α-glucans that can be synthesized.
研究领域:
食品科学
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生物化学
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酶工程
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DOI
高影响因子
JOURNAL OF DRUG DELIVERY SCIENCE AND TECHNOLOGY
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2025
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IF 4.9
Feifei Wang,Dianlong Jia,Qingcui Song,Xiaomeng Lu,Guozi Xia,Jun Li,Xiaodong Jia,Mingliang Gu,Fengjiao Yuan,Zhengping Wang,Zhangyong Xia
Resistance against tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced cell death of cancer cells is a major obstacle in clinical application of TRAIL. Bortezomib (BTZ) is the first-generation proteasome inhibitor to enhance TRAIL-mediated apoptosis and overcome tumor cell resistance to TRAIL, therefore we developed a novel nanodrug called LSBTZ-TRAIL to co-deliver TRAIL and BTZ via liposomes. TRAIL was coupled to the surface of the liposome by DSPE-PEG 2000 -NHS, while BTZ was encapsulated in the internal aqueous phase. LSBTZ-TRAIL was characterized with a particle size of 147.00 ± 1.300 nm, polydispersity index (PDI) of 0.056 ± 0.005 and zeta potential of -8.17 ± 0.600 mV. LSBTZ-TRAIL did not change the secondary structure of TRAIL, and LSBTZ-TRAIL was stable in 50% fetal bovine serum and tumor microenvironment. In vitro, the cell killing capacity of LSBTZ-TRAIL was 1.16-6.22 times that of TRAIL + BTZ. Mechanistically, LSBTZ-TRAIL induced apoptosis of tumor cells by up-regulating the death receptor (DR4/5) and activating the caspase signaling pathway. In vivo and in vitro uptake experiments, LS-TRAIL accumulated more in tumor cells or tissues than LS. In vivo , the antitumor effect of LSBTZ-TRAIL was 2.62 times that of TRAIL + BTZ, and prolonged the survival and blood circulation time of mice. In addition, safety evaluation showed that LSBTZ-TRAIL had no acute toxic effects in vivo . These results suggested that the liposomal co-delivery system of TRAIL and BTZ significantly enhance their antitumor effects and provide a new approach to overcome TRAIL-resistance.
研究领域:
肿瘤学
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药学
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分子生物学
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产品关联
DOI
高影响因子
Current Research in Microbial Sciences
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2024
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IF 4.8
Sirui Zhang,Jingdan Wang,Rong Yu,Haiping Liu,Shuyan Liu,Kai Luo,Jin'e Lei,Bei Han,Yanjiong Chen,Shaoshan Han,E Yang,Meng Xun,Lei Han
Although various mechanisms of carbapenem-resistance have been identified in the nosocomial pathogen Acinetobacter baumannii , the critical process of resistance evolution and the factors involved in are not well understood. Herein, we identified a universal stress protein Usp1413 which played an important role in adaptive resistance of A. baumannii to meropenem (MEM). Based on RNA-Seq and genome sequencing, Usp1413 was not only one of the most downregulated USPs, but also the bare one having mutation of tyrosine and glycine inserted at the site of 229-230 (YG229-230) under the stimulation of MEM. Deletion of Usp1413 resulted in increased MEM resistance. In addition, Usp1413 affected the bacterial abilities of biofilm formation and swarm motility, as well as helped A. baumannii response to various environmental stresses. These effects of Usp1413 were achieved by regulating its interaction proteins, within the functions of YigZ family protein, acetyltransferase, and SulP family inorganic anion transporter. The insertion mutation of YG229-230 influenced both the expression of interaction proteins and the phenotypes of bacteria. Finally, the promotor region of Usp1413 was convinced by point mutations. Overall, our findings identified the universal stress protein Usp1413 as a contributor involved in MEM adaptive resistance and responded to numerous environmental stresses. This study provides novel insights into the mechanism of universal stress proteins in participating antibiotic resistance, and affords a potential target for controlling drug resistance development in A. baumannii .
研究领域:
微生物学
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抗药性研究
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Bioresources and Bioprocessing
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2025
·Vol. 12
·期 1
·1-14
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IF 5.1
Ai Yulin,Huang Yusong,Zhao Hongru,Su Bingmei,Lin Juan
5-Hydroxytryptophan (5-HTP) is widely used as a natural remedy for sleep disorders. In terms of biosafety, bio-derived 5-HTP is preferred over chemically synthesized 5-HTP. However, the low titer of 5-HTP in the reported microbiological methods (< 10 g/L) limits the industrialization of 5-HTP biosynthesis. In the present study, a Trp-accumulating E. coli strain TRP1 was constructed by blocking the degradation path (Δ tnaA ), branching paths (Δ pheA , Δ tyrA ) and repression system (Δ trpR , Δ trpL ). Next, the hydroxylation module employing a phenylalanine hydroxylase mutant XcPAH W179F (XC2) coupled with an MH4 regenerating system (CvPCD-EcFolM system) was screened to convert L-Trp into 5-HTP. Protein engineering was performed on hydroxylase XC2 based on the molecular dynamics simulation of the enzyme-substrate complex, and the strain TRP1-XC4 harboring the triple-mutant XcPAH L98I/A129K/W179F (XC4) was able to produce 319.4 mg/L 5-HTP. Genome editing was carried out focused on accelerating product efflux (strengthening YddG) and increasing MH4 supply (strengthening FolM, FolE and FolX), resulting in a strain TRP5-XC4 to produce 13.9 g/L 5-HTP in 5 L fed-batch fermentation with a space-time yield of 0.29 g/L/h, which is the highest production and productivity record for 5-HTP biosynthesis. This study successfully provided an engineered strain and an efficient green method for the industrial synthesis of 5-HTP. Graphical
研究领域:
生物工程
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微生物学
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合成生物学
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DOI
PubMed
高影响因子
BIORESOURCE TECHNOLOGY
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2024
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IF 9.7
Yizhen Zhang,Yu-Cai He,Cuiluan Ma
Lignin is a plentiful and readily accessible renewable resource. Vanillylamine is a crucial raw material used to synthesize pharmaceuticals and high-value furan compounds that can be acquired by aminating lignin-derived vanillin (Van). However, effectually achieving the biocatalytic synthesis of vanillylamine has remained challenging. In this study, a dimethyl sulfoxide (DMSO)-H 2 O (1:9, vol/vol) bioreaction medium was constructed, and a recombinant E. coli ATA1012 carrying ω-transaminase from Caulobacter sp. D5 was used as the ω-transaminase biocatalyst to acquire the effectual biocatalytic synthesis of vanillylamine. Under optimized bioreaction conditions (37 ℃ and pH 7.5) by supplementary of isopropylamine (IPA) (Van/IPA = 1:5, mol/mol), 80–100 mM Van could be effectually converted by ATA1012 whole cells in DMSO-H 2 O (1:9, vol/vol) within 12 h, yielding 91.2 %–95.4 % vanillylamine, with >99 % selectivity. An efficient amination process was developed using ATA1012 with superior transaminase catalytic activity and substrate tolerance to effectively convert Van to vanillylamine in a DMSO-H 2 O medium.
研究领域:
生物技术
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生物化学
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有机化学
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药学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Ziqi Liang,Qiao Zhou,Yicheng Li,Xiaoqing Liu,Yiwen Shen,Jian Tian,Xiaolu Wang,Xing Qin,Yuan Wang,Huiying Luo,Xiaojun Yang,Bin Yao,Tao Tu
Cytochrome P450 enzymes (P450s) are promising candidates for the biosynthesis of 25-hydroxyvitamin D3 (25(OH)VD3). However, their industrial application is limited by challenges, such as low stability, inefficient catalysis, and uncoupling reactions. The construction of self-sufficient P450s offers a strategic solution to these limitations, but requires linker optimization to regulate interdomain conformational dynamics. In this study, we integrated whole-cell biocatalyst screening with systematic optimization of reaction conditions, including cosolvents, cell concentrations, and plasmid selection, to enhance catalytic performance. Under optimized conditions, the heme domain Vdh-K1 achieved a 91.6% conversion efficiency and was subsequently selected for chimeric enzyme assembly. By employing local energetic frustration analysis to evaluate protein flexibility and allosteric dynamics, we identified chimeric P450 variants with highly frustrated linkers. The optimal variant, VK1-CYP116B46-L21, exhibited improved thermostability, catalytic activity, and coupling efficiency, achieving a yield of 4.89 mM (1.96 g/L) 25(OH)VD3 in Escherichia coli whole-cell catalysis─the highest reported yield to date. This work underscores the utility of computational frustration analysis in refining linker dynamics for multidomain enzymes and establishes a scalable, cost-effective framework to advance P450s systems for industrial biosynthesis of high-value compounds.
研究领域:
生物化学
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工业生物技术
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酶工程
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产品关联
DOI
PubMed
高影响因子
Processes
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2024
·Vol. 12
·期 9
·2006
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IF 2.8
Wei He,Benshun Chen,Bin Yin,Jianren Ye,Yucai He
Simvastatin is a widely used statin medication that is commonly prescribed to lower cholesterol levels and reduce the risk of cardiovascular events. It is marketed under the brand name Zocor and is known for its effectiveness in treating high cholesterol and managing cardiovascular disease. Monacolin J is an important precursor used to synthesize simvastatin and is mainly produced by chemical methods in industry. Here, monacolin J was synthesized through an enzymatic method under optimized reaction conditions. One recombinantEscherichia coliBL21 (DE3) strain containing lovastatin hydrolase (encoded by CDV55_102090) fromAspergillus turcosuswas constructed, which effectively transformed 100 g/L of lovastatin to monacolin J within 3.5 h at pH 8.0 and 30 °C, with a conversion rate of >99.8%. Furthermore, the T5010, the temperature at which the residual activity was half of the initial enzymatic activity after 10 min of heat treatment, was >50 °C, indicating the tremendous potential of this bioprocess for synthesizing monacolin J at an industrial scale.
研究领域:
生物化学
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制药工程
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工业微生物学
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DOI
高影响因子
JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
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2025
·
IF 3.1
Lin Ge,Yu Xia,Wenxin Xu,Ruobing Jia,Tingting Zhang
Gastrodin, the primary bioactive constituent of Gastrodia elata , possesses numerous remarkable pharmacological properties. In this investigation, UDP-glycosyltransferase from Rauvolfia serpentina (RsUGT) was expressed, subsequently purified and characterized. The maximum yield of the enzyme was 17.57 mU/ml and possessed a relative molecular weight of approximately 77.7 kDa. Utilizing GST affinity resin, RsUGT was purified 20.8-fold, with an overall recovery rate of 58.6% and specific activity of 79.2 mU/mg. The optimal temperature and pH for RsUGT was identified as 40°C and 10.0, respectively. Notably, 2% DMSO could increase the RsUGT activity by 12.15%. The Michaelis-Menten constants K M and V max were determined to be 0.50mM and 171.60 mU/mg. By optimizing the conditions for the enzymatic biosynthesis of gastrodin by RsUGT, the highest gastrodin production was 285.35 mg/l, accompanied by a molar conversion rate of 99.67%. In addition, the conditions of gastrodin biosynthesis by recombinant strain BL-RsUGT were also studied. The highest gastrodin production was 225.99 mg/l, and the corresponding p HBA conversion rate was 98.00%. These findings confirmed the promising potential of RsUGT in the production of gastrodin.
研究领域:
植物学
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药理学
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生物化学
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产品关联
DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
Ting Wu,Huashan Sun,Wenyao Wang,Bin Xie,Zhengjie Wang,Jianqi Lu,Anming Xu,Weiliang Dong,Jie Zhou,Min Jiang
Enzymatic PET recycling has emerged as a promising green solution in addition to mechanical recycling, but low soluble expression levels of the inherently hydrophobic PET hydrolases hinder large-scale applications. Here, we propose a novel strategy for enhanced production of FastPETase in Escherichia coli using co-expression of molecular chaperones from Ideonella sakaiensis . Co-expression of cognate DnaK and DnaJ chaperones significantly increased soluble FastPETase expression (up to 2.5-fold), surpassing commercial chaperone plasmids. Furthermore, a combinatorial approach employing co-expression of DnaK/DnaJ chaperones and fusion of FastPETase with the VNp6-tag significantly boosted FastPETase secretion, yielding over 2 g/L of target protein in a 5-l bioreactor. Notably, the crude FastPETase in fermentation broth displayed comparable PET hydrolysis effects to the purified enzyme. This work not only provides new insights into the process of chaperones in protein folding but also suggests a novel and efficient strategy for producing recombinant proteins.
研究领域:
生物化学
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环境科学
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蛋白质工程
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DOI
PubMed
高影响因子
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2024
·
IF 7.7
Tengfei Xiong,Mingyue Yu,Mei Li,Christopher A. Reilly,Xinghui Qiu
Capsaicin and dihydrocapsaicin are the principal pungent compounds in hot peppers. The generalist Helicoverpa armigera and the specialist H. assulta are two of the few insects that can feed on hot pepper fruits. To further understand the mechanisms behind physiological adaptation to capsaicinoids in the two agricultural pests, we investigated the transcriptional response and metabolism capacity of CYP6B6 orthologs from the two closely related species to the capsaicinoids. The results showed that the expression of CYP6B6 was either weakly or moderately induced by capsaicin in a tissue-specific and dose-dependent manner. CYP6B6 induction to greater extent was observed in the larval midguts of the cotton bollworms, indicating that inducible defense is more robust in the generalist. The two CYP6B6 enzymes were able to efficiently metabolize both capsaicin and dihydrocapsaicin, and HassCYP6B6 exhibit a slightly higher activity towards capsaicin than HarmCYP6B6, as evaluated by the specific activity and Vmax values. Multiple known and yet to be elucidated metabolites were detected in the reactions catalyzed by CYP6B6s, with dehydrogenation at the terminal carbon followed by N-macrocyclization and aliphatic hydroxylation being the dominant pathways. The data support the hypothesis that these two Helicoverpa species have well adapted to capsaicinoids via their high P450-mediated detoxification ability.
研究领域:
昆虫学
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分子生物学
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植物与昆虫互作
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产品关联
DOI
PubMed
高影响因子
3 Biotech
·
2024
·Vol. 15
·期 1
·1-8
·
IF 2.6
Feng Xinming,Cui Xinyu,Wang Kun,Liu Juanjuan,Meng Dongdong
6-Phosphogluconate dehydrogenases (6PGDHs) are widely existing as reduced cofactor (NADH/NADPH) regeneration biocatalysts. Herein, a thermostable 6PGDH from Hydrogenobacter thermophilus (Ht6PGDH) was overexpressed in Escherichia coli and enzymologically characterized. Ht6PGDH exhibited exceptional stability and catalytic activity under high-temperature conditions, with an optimum temperature of 85 °C and the ability to maintain high activity for prolonged periods at 70 °C, which could be purified through a one-step heat treatment. Moreover, Ht6PGDH exhibited a preference for NAD + with a K m value of 0.4 mM and a k cat value of 28.6 s⁻ 1 , demonstrating a significant preference over NADP + . These properties render Ht6PGDH a potentially valuable enzyme for high-temperature bioconversion and in vitro synthetic biosystems. Additional research showed that Ht6PGDH excelled in the regeneration of NADH, achieving efficient lactate production when integrated into an in vitro synthetic biosystem containing lactate dehydrogenase (LDH). Furthermore, the cascade reaction of Ht6PGDH with glucose-6-phosphate dehydrogenase (G6PDH) was explored for NADH regeneration using starch as the substrate, further validating its potential application in complex biosynthetic systems.
研究领域:
生物化学
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酶工程
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合成生物学
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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）