BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
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2024
·
IF 3.2
Qian Tan,Linbo Gou,Tai-Ping Fan,Yujie Cai
Clostridium aceticum DSM1496 is an acid-resistant strain in which ornithine decarboxylase (ODC) plays a crucial role in acid resistance. In this study, we expressed ODC derived from C. aceticum DSM1496 in Escherichia coli BL21 (DE3) and thoroughly examined its enzymatic properties. The enzyme has a molecular weight of 55.27 kDa and uses pyridoxal-5′-phosphate (PLP) as a coenzyme with a K m = 0.31 mM. ODC exhibits optimal activity at pH 7.5, and it maintains high stability even at pH 4.5. The peak reaction temperature for ODC is 30°C. Besides, it can be influenced by certain metal ions such as Mn 2+ . Although l -ornithine serves as the preferred substrate for ODC, the enzyme also decarboxylates l -arginine and l -lysine simultaneously. The results indicate that ODC derived from C. aceticum DSM1496 exhibits the ability to produce putrescine, cadaverine, and agmatine through decarboxylation. These polyamines have the potential to neutralize acid in an acidic environment, facilitating the growth of microorganisms. These significant findings provide a strong basis for further investigation into the acid-resistant mechanisms contributed by ODC.
研究领域:
微生物学
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生物化学
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产品关联
DOI
PubMed
高影响因子
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
·
2015
·Vol. 19
·期 6
·965-972
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IF 1.113
Li Naiqiang,Chou Howard,Yu Lijun,Xu Yan
Cadaverine, as a biogenic amine, is an important platform chemical for the production of industrial polymers, such as polyamides, polyurethanes, and nylon. Previous efforts focused on the bio-based production of cadaverine using two lysine decarboxylases of Escherichia coli CadA and LdcC. In this study, we report the biotransformation of cadaverine using a lysine decarboxylase from Klebsiella oxytoca . Codon optimization of the gene encoding this enzyme was carried on for the heterologous expression in E. coli , which led to a system that converted more than 24% lysine-HCl to cadaverine compared to the same system expressing CadA. The system was further optimized by using three different inducible promoters to control the expression of lysine decarboxylase gene of K. oxytoca in E. coli . The final optimized system converted lysine-HCl to cadaverine at a conversion rate of 0.133%/min/g. When the optimized system described above is coupled to an industrial process, the combined process has the potential to produce cadaverine with high conversion efficiency (46%) from sugar.
研究领域:
生物化学
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合成生物学
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工业微生物学
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DOI
高影响因子
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
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2022
·Vol. 38
·期 5
·1-8
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IF 4.253
Li Yudi,Chen Guiying,Ge Fanglan,Dang Ting,Ren Yao,Zeng Bei,Li Wei
γ-aminobutyric acid (GABA) has various physiological functions and is widely used in medicine, food, and other fields. Glutamate decarboxylase (GAD) is a key enzyme that catalyzes the decarboxylation of L-glutamate to synthesize GABA. However, the industrial application of microorganism-derived GAD is limited by its rapid loss of enzymatic activity with pH approaching neutrality. In this study, a novel glutamate decarboxylase, GAD MSM, from Mycobacterium smegmatis was overexpressed and purified. On the basis of homologous modeling and substrate molecular docking, several GAD MSM mutants were constructed, and their enzymatic properties were analyzed. The results showed that the optimal pH of wild-type GAD MSM is 5.4; at pH 6.2, 22.8% enzymatic activity was retained. The T211I replacement in GAD and C-terminal deletion mutant GAD MSMΔC showed relatively high catalytic activity in a pH range of 5.0–7.0. The V max and K m values of GAD MSMΔC were 14.69 and 5.70, respectively, at pH 5.5, and 9.87 and 6.17, respectively, at pH 7.0. Compared with the wild-type GAD, GAD MSMΔC maintained higher affinity and enzymatic activity of the substrate, maintaining 78.5% of the highest enzymatic activity even at pH 7.0, which is the highest reported activity retention for GAD under neutral pH condition. Therefore, GAD MSMΔC can be used for the transformation of high-yielding strains and industrial production of GABA.
研究领域:
微生物学
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酶工程
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生物化学
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产品关联
DOI
PubMed
高影响因子
CHINESE JOURNAL OF CATALYSIS
·
2022
·
IF 12.92
Yunjian Ma,Xuanru Zhong,Bin Wu,Dongming Lan,Hao Zhang,Frank Hollmann,Yonghua Wang
Hydrocarbons are essential base chemicals as energy carriers and starting materials for chemical manufacture. So-called fatty acid photodecarboxylases (FAPs) represent interesting catalysts for the conversion of natural fatty acids into hydrocarbons thereby giving access to alkanes from renewable feedstock. Today, however, only few FAPs are known. In the current study we report a new FAP from the marine organism Micractinium conductrix ( Mc FAP). In contrast to currently known FAPs Mc FAP exhibits high catalytic activity towards short and medium fatty acids. Recombinant expression and basic biochemical characterisation of this new member of the FAP family is reported.
研究领域:
生物化学
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酶学
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可再生能源
✓ 已验证
DOI
高影响因子
BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY
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2020
·Vol. 68
·期 2
·381-389
·
IF 1.638
Dawei You,Yue Feng,Can Wang,Chengtao Sun,Yao Wang,Degang Zhao,Guoyin Kai
Tryptophan decarboxylase (TDC, EC 4.1.1.28) catalyzes tryptophan decarboxylation to form tryptamine through the cofactor pyridoxal-5′-phosphate (PLP), a crucial stage in the production of the terpenoid indole alkaloids like camptothecin (CPT). A new gene encoding TDC was identified from the CPT-producing plant Ophiorrhiza pumila by transcriptome analysis, termed OpTDC2. It contained a 1,536 bp open reading frame that encodes a 511 amino acid protein with a molecular mass of 57.01 kDa and an isoelectric point of 6.39. Multiple sequence alignment and phylogenetic tree analysis showed the closest similarity (85%) with the TDC from Mitragyna speciosa . Moreover, the highest expression of OpTDC2 was observed in the O. pumila root. To achieve high-efficiency expression of OpTDC2 in Escherichia coli , we fused the TF tag onto the N-terminal of the OpTDC2. Optimum enzymatic activity was observed at 45 °C, pH 8 and cofactor concentration of 0.1 mM. The catalytic reaction was strongly inhibited by metal ions of Cu 2+ , Zn 2+ , and Fe 2+ . The l -tryptophan was particularly catalyzed compared with d -tryptophan. Besides, the K m and k cat of the OpTDC2 were 1.08 mM and 0.78 Sec −1 , respectively. The results provided information on new functional OpTDC2 that might be used in synthetic biology for the enhanced biosynthesis of CPT in O. pumila .
研究领域:
植物学
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生物化学
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分子遗传学
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DOI
PubMed
高影响因子
Bioresource Technology Reports
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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
✓ 已验证
DOI
ACS Sustainable Chemistry & Engineering
·
2021
·
IF 8.198
Jiali Mi,Simin Liu,Haishan Qi,Jie Huang,Xiang Lan,Lei Zhang
Cadaverine, a diamine monomer, broadly exists in living organisms participating in metabolism processes. As a promising substitute of 1,6-diaminohexane, cadaverine can polymerize with dicarboxylic acids and yield biobased polyamides, polyamide (PA) 5×, showing ecofriendly and excellent mechanical properties in the fields of electronics, automobile, material, storage, and others. Due to the increasing attentions on environment problems, biopolyamide is expected to be an ideal and green material to substitute conventional chemistry polyamide. This review summarizes the properties, potential applications, and production strategies about cadaverine and mainly focuses on the recent developments by cellular engineering Escherichia coli and Corynebacterium glutamicum as main workhorses. In addition, the key enzyme lysine decarboxylase decorated by means of immobilization and mutation to efficiently catalyze lysine and its catalysis mechanisms are also discussed. In order to achieve industrial applications, the indispensable steps of separation and purification are described as well and perspectives for cadaverine manufacturing from renewable resources are further provided.
研究领域:
生物化学
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微生物学
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合成生物学
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产品关联
DOI
高影响因子
JOURNAL OF CHEMICAL TECHNOLOGY AND BIOTECHNOLOGY
·
2020
·Vol. 95
·期 5
·1542-1549
·
IF 2.75
Yuxuan Liu,Yunxin Zheng,Hao Wu,Wei Zhang,Tianyu Ren,Shengping You,Wei Qi,Rongxin Su,Zhimin He
BACKGROUND Cadaverine is a natural polyamine that can replace fossil-derived hexamethylene diamine to produce bio-based polyamide. Despite huge developments in cadaverine bioproduction, an econo"mically attractive purification process is a highly important part of the production of high-purity cadaverine. RESULTS This study utilized l -lysine hydrochloride as a substrate for the production of high-purity cadaverine via a litre-scale integrated process incorporating fermentation, bioproduction, deprotonation, extraction and rectification. First, 8.24 U mg –1 lysine decarboxylase activity and 205 g L –1 cadaverine with a yield of 91.61% were achieved through the fermentation of low-cost industrial culture by engineered Escherichia coli and the bioproduction of cadaverine by lysine decarboxylase. Secondly, four integrated purification methods – deprotonation-evaporation, pH adjustment-deprotonation-evaporation, deprotonation-extraction-evaporation and deprotonation-extraction-rectification – were explored, and the conditions of each unit operation in the chosen method (deprotonation-extraction-rectification) were optimized. Then the optimized method was applied to a 2-L purification system, resulting in 99% purity of cadaverine with a yield of 87.47%. Furthermore, the n -butanol recovery ratio reached 91.25%, effectively avoiding the waste of resources and environmental pollution, and thus reducing purification costs. CONCLUSION The proposed experimental approach proves that this integrated process leads to the production of high-purity cadaverine. In the 2-L-scale integrated process, 358.64 g cadaverine of 99% purity was produced at a cost of US$3.99 (US$1.11 per 100 g), which is far below its market price (US$784.71 per 100 g). This integrated process provides a potential way for the industrial-scale production of bio-based cadaverine. © 2020 Society of Chemical Industry
研究领域:
生物工程
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化学工业
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产品关联
DOI
高影响因子
Journal of Environmental Chemical Engineering
·
2023
·
IF 7.7
Feng Li,Ao Xia,Xiaobo Guo,Wuyuan Zhang,Yun Huang,Xianqing Zhu,Xun Zhu,Qiang Liao
Fatty acid photodecarboxylase, recently found in Chlorella variabilis NC64A ( Cv FAP), could efficiently catalyze C16-C18 fatty acids into C1-shortened hydrocarbons under visible light. This provides an alternative approach for hydrocarbon fuels production in mild conditions compared with hydrotreating of oils. In this study, a cascade system integrating lipase hydrolysis and Cv FAP decarboxylation was demonstrated for the efficient production of hydrocarbon fuels from waste oils. The continuous Cv FAP-catalyzed decarboxylation characteristics and performance of free fatty acids derived from waste oil were investigated for the first time. A hydrocarbon fuel production rate of 18.4 mM/h was obtained at a residence time of 50 min. The regulation of pH value from 6.8 to 8.5 significantly improved the activity of Cv FAP, thereby obtaining an optimal conversion of 61.2%. The precipitation of palmitic acid caused a low conversion (37.9%) compared with the other fatty acids, further inhibiting hydrocarbon production. In such a cascade system, various oils can be converted to hydrocarbon fuels with a concentration of 8.97–15.3 mM. A high energy production rate of 204.3 kJ/L/h was achieved, which exhibited the advancement in high-quality biofuel production.
研究领域:
生物化学
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可再生能源
·
环境科学
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产品关联
DOI
高影响因子
JOURNAL OF BIOTECHNOLOGY
·
2021
·
IF 3.307
Jingbai Wen,Jie Bao
Fermentative production of γ-aminobutyric acid by the glutamate overproducing Corynebacterium glutamicum from cheap sugar feedstock is generally regarded as one of the most promising methods to reduce the production cost. However, the intracellularly expressed glutamate decarboxylase in C. glutamicum often showed feeble catalysis activity to convert glutamate into γ-aminobutyric acid. Here we tried to secretory express glutamate decarboxylase to achieve efficient extracellular decarboxylation of glutamate, thus improving the γ-aminobutyric acid production by C. glutamicum . We first tested glutamate decarboxylases from different sources, and the mutated glutamate decarboxylase GadBmut from E. coli with better catalytic performance was selected. Then, a signal peptide of the SEC translocation pathway directed the successful secretion of glutamate decarboxylase in C. glutamicum . The extracellular catalysis by secreted glutamate decarboxylase increased the γ-aminobutyric acid generation by three-fold, compared with intracellular catalysis. Enhancing glutamate decarboxylase expression and decreasing γ-aminobutyric acid degradation further increased γ-aminobutyric acid production by 39 %. The fed-batch fermentation of the engineered C. glutamicum strain reached the record high titer (77.6 ± 0.0 g /L), overall yield (0.44 ± 0.00 g/g glucose), and productivity (1.21 ± 0.00 g/L/h). This study demonstrated a unique design of extracellular catalysis for efficient γ-aminobutyric acid production by C. glutamicum .
研究领域:
微生物学
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生物技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Advanced Sustainable Systems
·
2025
·
IF 6.1
Jianle Zhou,Xiongliang He,Yanyan Ye,Zhuoxin Zhuang,Xingyu Tang,Junpeng Zhao,Yunjian Ma,Yonghua Wang
Fatty acid photodecarboxylase (FAP) plays a crucial role in the green production of biofuel and other valuable biochemicals. However, the reusability of immobilized FAP has been limited due to inadequate durability. Here, a porous, translucent chitosan hydrogel sphere carrier functionalized with cyclic carbonate group to enhance the reusability of immobilized FAP is presented. Based on the arrangement of basic amino acid residues on the surface of FAP, bis(cyclic carbonate) containing a flexible chain from CO 2 is designed and synthesized. This compound is used to modify porous hydrogels obtained via a template-etching process. FAP is then covalently immobilized within the hydrogel framework through a reaction with the remaining cyclic carbonate groups, as evidenced by quartz crystal microbalance analysis. The modified porous hydrogel carrier, PH3-BC-II, significantly improves the activity of FAP, achieving a maximum conversion of 70.0%, with the enzyme loading of 125.3 mg g −1 (dry carrier). Furthermore, PH3-BC-II retains >50% of its initial activity after eight consecutive reaction cycles (total runtime of 24 h) at high fatty acid substrate concentrations. This study provides an effective strategy for constructing stable immobilized (photo)enzymes from sustainable materials.
研究领域:
生物化学
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材料科学
·
酶工程
✓ 已验证
产品关联
DOI
高影响因子
BIOCHEMICAL ENGINEERING JOURNAL
·
2025
·
IF 3.8
Yan Wang,Bo Ding,Qingqing Bao,Yunying Zhao,Guohui Li,Yu Deng
Agmatine is an important basic drug and nutritional supplements, which exhibits significant physiological activities. Traditional methods, such as whole-cell catalysis, require substantial pyridoxal 5′-phosphate (PLP) addition, which accounts for 15 % of production costs. In this study, we utilized purified arginine decarboxylase (AdiA-M2) that leverages the intrinsic PLP cofactor, without the need for additional PLP supplementation, and ZIF-8 was employed to enhance the pH tolerance. The successful incorporation of AdiA-M2 into the ZIF-8 structure was confirmed through scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). The immobilized arginine decarboxylase demonstrated markedly enhanced performance under alkaline conditions, along with improved thermal stability and storage activity compared to its free enzyme counterpart. Specifically, the immobilized AdiA@ZIF-8 system achieved an initial conversion rate of 95.16 %, after five recycles the titer attained 266.69 g/L without the extra PLP addition, which is 1.67 times higher than the best current production approach. The adopted enzyme-catalysis system eliminated the need for additional cofactors, thereby lowering costs and providing valuable insights for the industrial production of agmatine.
研究领域:
生物化学
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酶工程
·
制药工程
✓ 已验证
产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）