INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
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2024
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IF 7.7
BingBing Wang,Jian-Zhong Xu,Shuai Liu,Zhi-Ming Rao,Wei-Guo Zhang
L-Tryptophan hydroxylation catalyzed by tryptophan hydroxylase (TPH) presents a promising method for synthesizing 5-hydroxytryptophan (5-HTP), yet the limited activity of wild-type human TPH2 restricts its application. A high-activity mutant, MT10 (H318E/H323E), was developed through semi-rational active site saturation testing (CAST) of wild-type TPH2, exhibiting a 2.85-fold increase in k cat / K m over the wild type, thus enhancing catalytic efficiency . Two biotransformation systems were developed, including an in vitro one-pot system and a Whole-Cell Catalysis System (WCCS). In the WCCS, MT10 achieved a conversion rate of only 31.5 % within 32 h. In the one-pot reaction, MT10 converted 50 mM L-tryptophan to 44.5 mM 5-HTP within 8 h, achieving an 89 % conversion rate, outperforming the M1 (NΔ143/CΔ26) variant. Molecular dynamics simulations indicated enhanced interactions of MT10 with the substrate, suggesting improved binding affinity and system stability. This study offers an effective approach for the efficient production of 5-HTP.
研究领域:
生物化学
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分子生物学
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合成生物学
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DOI
PubMed
高影响因子
JOURNAL OF LIGHTWAVE TECHNOLOGY
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2025
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IF 4.8
Xiu Du,Lingxin Kong,Mingxing Liu,Yu Zhong,Kuihua Ding
In this paper, a fiber optic tryptophan enantiomers chiral sensor based on metal-organic frameworks (MOFs) is proposed. Two kinds of MOFs with different chiral microenvironments were coated on the surface of U-shaped sensor and helical long-period grating (HLPG), and the two sensors achieved specific identification and quantitative sensing of L-Trpptophan (L-Trp) and D-Trpptophan (D-Trp) in racemic mixture solution. The two MOFs have different adsorption effects on L-Trp and D-Trp, which makes the two sensors have different sensitivity. The results showed that the sensitivity of the sensor was -24.34 nm/μM (experimental limit of detection (LOD) = 2.5 nM) for L-Trp and 11.579 nm/μM (experimental LOD = 5.2 nM) for D-Trp. The fiber Bragg grating (FBG) is used as a temperature sensor to compensate the temperature of the sensing system. Chiral detection of tryptophan enantiomers at different temperatures can be achieved by constructing a three parameters matrix. Research has shown that the sensor has a satisfactory application prospect.
研究领域:
生物化学
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光学传感
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材料科学
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DOI
高影响因子
TALANTA
·
2018
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IF 4.244
Peng Lei,Ying Zhou,Guomei Zhang,Yan Zhang,Caihong Zhang,Shasha Hong,Yajuan Yang,Chuan Dong,Shaomin Shuang
Construction of convenient and effective method for enantiomer identification is of vital significance for biochemistry and medical science. Herein, we design an effective sensor for chiral recognition of tryptophan (Trp) enantiomers, and self-assembly of Cu 2+ -modified β-cyclodextrin on poly- L -arginine/multi-walled carbon nanotubes (Cu-β-CD/PLA/MWCNTs) is studied. Meanwhile, Cu 2+ acts as a cap to prevent the release of the high energy water and compel Trp enantiomer into the smaller opening of β-cyclodextrin. Recognition of L -Trp is accomplished by the formation of hydrogen bonds between the amino of L -Trp and the high energy water confined in cavity of Cu-β-CD. Compared with D -Trp, the sensor exhibits favorable chiral recognition toward L -Trp with a separation coefficient of 3.37. And the chiral sensor presents admirable enantiomers determination with excellent sensitivity, providing a good linear correlation in the range of 1 × 10 −6 M~5.5 × 10 −5 M, and the detection limit can reach 3.3 × 10 −7 M (S/N = 3). Besides, the proposed sensor has been able to predict the percentage of D -Trp in the racemic mixture, suggesting its potential applications in the enantiomer recognition field.
研究领域:
生物化学
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医学科学
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分析化学
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产品关联
DOI
PubMed
高影响因子
ENZYME AND MICROBIAL TECHNOLOGY
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2023
·
IF 3.4
Feifei Song,Tao Gu,Lin Zhang,Jiaxing Zhang,Shengping You,Wei Qi,Rongxin Su
5-Hydroxytryptophan (5-HTP) is a chemical precursor of serotonin, which synthesizes melatonin and serotonin in animals and regulates mood, sleep , and behavior. Tryptophan hydroxylase (TPH) uses tetrahydrobiopterin (BH4) as a cofactor to hydroxylate L -tryptophan ( L -Trp) to 5-HTP, and the low catalytic activity of TPH limits the rate of hydroxylation of L -Trp. In this study, the catalytic mechanism and structural features of L -Trp-TPH1-BH4 were investigated, and the catalytic activity was improved using a rational design strategy. Then the S337A/F318Y beneficial mutation was obtained. Molecular dynamics simulations showed that the S337A/F318Y mutant formed a salt bridge with TPH1 while forming an additional hydrogen bond with the substrate indole ring, stabilizing the indole ring and enhancing the binding affinity of the variant to L -Trp. As a result, the yield of 5-HTP was increased by 2.06-fold, resulting in the production of 0.91 g/L of 5-HTP. The rational design of the TPH structure to improve the hydroxylation efficiency of L -Trp offers the prospect of green production of 5-HTP.
研究领域:
生物化学
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分子生物学
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代谢工程
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产品关联
DOI
PubMed
高影响因子
Molecular Catalysis
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2023
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IF 4.6
BingBing Wang,Shuai Liu,HaiDe Wang,JianZhong Xu,WeiGuo Zhang
5-Hydroxytryptophan (5-HTP) is a precursor of serotonin. The increasing demand for 5-HTP is driving the search for a more efficient and cost-effective production method. In this study, a new in vitro multi-enzyme cascade system (MECCS) was developed to convert L -tryptophan (L-trp) to 5-HTP in a single container while achieving the cycling and regeneration of cofactors and coenzymes. The rate-limiting enzyme tryptophan hydroxylase (TPH) was truncated to improve the overall efficiency of the cascade. After screening, the mutant M1 (NΔ143/CΔ26) showed excellent enzymatic performance, with a 2.3-fold increase in catalytic activity. Molecular dynamics simulations showed that the increase in active center binding force and the improvement of substrate tunnel in M1 improved the overall catalytic efficiency. In addition, the rigidity enhancement of the ligand binding pocket surrounding the region improved the thermal stability of the enzyme. Under optimized conditions, the MECCS converted 50 mM L -trp to 36.9 mM 5-HTP, with a conversion rate of 73.8%. This study provides an alternative pathway for the efficient production of 5-HTP.
研究领域:
生物技术
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生物化学
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酶学
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药学
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产品关联
DOI
高影响因子
MICROCHIMICA ACTA
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2018
·Vol. 185
·期 4
·1-9
·
IF 5.705
Song Jinyi,Yang Chengcheng,Ma Jiao,Han Qian,Ran Peiyao,Fu Yingzi
An electrochemical chiral multilayer nanocomposite was prepared by modifying a glassy carbon electrode (GCE) via opposite-charge adsorption of amino-modified β-cyclodextrin (NH 2 -β-CD), gold-platinum core-shell microspheres (Au@Pts), polyethyleneimine (PEI), and multi-walled carbon nanotubes (MWCNTs). The modified GCE was applied to the enantioselective voltammetric determination of tryprophan (Trp). The Au@Pts enable an effective immobilization of the chiral selector (NH 2 -β-CD) and enhance the electrochemical performance. Scanning electron microscopy, transmission electron microscopy, UV-vis spectroscopy, FTIR and electrochemical methods were used to characterize the nanocomposite. Trp enantiomers were then determined by differential pulse voltammetry (DPV) (with a peak potential of +0.7 V vs. Ag/AgCl). The recognition efficiency was expressed by an increase in peak height by about 32% for DPV determinations of L-Trp compared to D-Trp in case of a 5 mM Trp solution of pH 7.0. Response was linear in the 10 μM to 5.0 mM concentration range, and the limits of detection were 4.3 μM and 5.6 μM with electrochemical sensitivity of 43.5 μA·μM −1 ·cm −2 and 34.6 μA·μM −1 ·cm −2 for L-Trp and D-Trp, respectively (at S/ N = 3).
Schematic of an electrochemical chiral multilayer nanocomposite composed of multi-walled carbon nanotubes (MWCNTs), polyethyleneimine (PEI), gold-platinum core-shell microspheres (Au@Pt) and amino-modified β-cyclodextrin (NH 2 -β-CD). It was prepared by modifying a glassy carbon electrode (GCE) for enantioselective voltammetric determination of tryptophan (Trp) enantiomers.
研究领域:
分析化学
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生物化学
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DOI
PubMed
高影响因子
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
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2020
·Vol. 104
·期 6
·2481-2488
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IF 3.53
Xu Da,Fang Mengjun,Wang Haijiao,Huang Lei,Xu Qinyang,Xu Zhinan
5-Hydroxytryptophan (5-HTP) is the precursor of the neurotransmitter serotonin and has been used for the treatment of various diseases such as depression, insomnia, chronic headaches, and binge eating associated obesity. The production of 5-HTP had been achieved in our previous report, by the development of a recombinant strain containing two plasmids for biosynthesis of L-tryptophan (L-trp) and subsequent hydroxylation. In this study, the L-trp biosynthetic pathway was further integrated into the E. coli genome, and the promoter strength of 3-deoxy-7-phosphoheptulonate synthase, which catalyzes the first step of L-trp biosynthesis, was engineered to increase the production of L-trp. Hence, the 5-HTP production could be manipulated by the regulation of copy number of L-trp hydroxylation plasmid. Finally, the 5-HTP production was increased to 1.61 g/L in the shaking flasks, which was 24% improvement comparing to the original producing strain, while the content of residual L-trp was successfully reduced from 1.66 to 0.2 g/L, which is beneficial for the downstream separation and purification. Our work shall promote feasible progresses for the industrial production of 5-HTP.
研究领域:
生物工程
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微生物学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Microbial Cell Factories
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2022
·Vol. 21
·期 1
·1-9
·
IF 6.352
Wang Yingying,Chen Xueman,Chen Qiaoyu,Zhou Ning,Wang Xin,Zhang Alei,Chen Kequan,Ouyang Pingkai
Background l-Tryptophan (l-Trp) derivatives such as 5-hydroxytryptophan (5-HTP) and 5-hydroxytryptamine (5-HT), N-Acetyl-5-hydroxytryptamine and melatonin are important molecules with pharmaceutical interest. Among, 5-HT is an inhibitory neurotransmitter with proven benefits for treating the symptoms of depression. At present, 5-HT depends on plant extraction and chemical synthesis, which limits its mass production and causes environmental problems. Therefore, it is necessary to develop an efficient, green and sustainable biosynthesis method to produce 5-HT.Results Here we propose a one-pot production of 5-HT from l-Trp via two enzyme cascades for the first time. First, a chassis cell that can convert l-Trp into 5-HTP was constructed by heterologous expression of tryptophan hydroxylase from Schistosoma mansoni (SmTPH) and an artificial endogenous tetrahydrobiopterin (BH4) module. Then, dopa decarboxylase from Harminia axyridis (HaDDC), which can specifically catalyse 5-HTP to 5-HT, was used for 5-HT production. The cell factory, E. coli BL21(DE3)△tnaA/BH4/HaDDC-SmTPH, which contains SmTPH and HaDDC, was constructed for 5-HT synthesis. The highest concentration of 5-HT reached 414.5 ± 1.6 mg/L (with conversion rate of 25.9 mol%) at the optimal conditions (substrate concentration,2 g/L; induced temperature, 25℃; IPTG concentration, 0.5 mM; catalysis temperature, 30℃; catalysis time, 72 h).Conclusion sThis protocol provided an efficient one-pot method for converting. l-Trp into 5-HT production, which opens up possibilities for the practical biosynthesis of natural 5-HT at an industrial scale.
研究领域:
生物技术
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药学
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代谢工程
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合成生物学
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产品关联
DOI
PubMed
高影响因子
Frontiers in Plant Science
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2022
·
IF 6.627
Qiao Chong,Chen Fei,Liu Zhan,Huang Tianfang,Li Wei,Zhang Guolin,Luo Yinggang
Tryptophan decarboxylases (TDCs) are a group of pyridoxal 5′-phosphate-dependent enzymes involved in the enzymatic conversion of tryptophan into tryptamine, a critical biogenic amine. We herein mined and cloned a TDC-encoding gene, CaTDC3, from camptothecin-producing plant Camptotheca acuminata. The intact CaTDC3 was heterologously overexpressed in Escherichia coli and the recombinant CaTDC3 was purified to homogeneity. High-performance liquid chromatography (HPLC)-diode array detector (DAD) and high resolution mass spectrometry (HRMS) data analyses of the CaTDC3-catalyzed reaction mixture confirmed the catalytically decarboxylative activity of CaTDC3. CaTDC3 shows strict stereoselectivity for L-tryptophan. Homology modeling and molecular docking implied CaTDC3’s recognition of L-tryptophan derivatives and analogs. Substrate scope investigations revealed that the appropriate substituent groups on the indole ring, i.e., hydroxylated and halogenated L-tryptophans, could be recognized by CaTDC3 and the decarboxylation reactions generated the corresponding tryptamines. The Cβ -methyl-L-tryptophans were decarboxylated by CaTDC3 efficiently. 1-Thio-L-tryptophan, the NH group of the indole ring replaced by an S atom, could be decarboxylated by CaTDC3. CaTDC3 catalyzed the decarboxylation of 7-aza-L-tryptophan, an N displacement of the C on the aromatic ring, to afford 7-aza-tryptamine. L-Kynurenine, an L-tryptophan degradation product, could be decarboxylated by CaTDC3. The present works uncover a catalytically promiscuous TDC and the TDC is a versatile decarboxylase in synthetic biology for specialized pharmaceutically important substances.
研究领域:
生物化学
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酶学
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药理学
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合成生物学
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产品关联
DOI
PubMed
高影响因子
ACS Sustainable Chemistry & Engineering
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2022
·
IF 9.224
Yaxuan Qian,Zhuoran Jiang,Yan Long,Houguo Fei,Heng Song,Cunlan Guo
Electrochemistry enables numerous redox enzymatic reactions with high efficiency. Nitroaromatic molecules with diverse biological properties have been extensively applied in medicine. New strategies are continuously required for the nitration of aromatic molecules with improved yields and selectivity. Here, we report a reaction by combining electrochemistry and the enzyme ferredoxin-TxtE to drive the nitration of l-tryptophan (l-Trp). With methyl viologen as the electron mediator, such an electroenzymatic approach avoids the use of expensive β-nicotinamide adenine dinucleotide phosphate (NADPH) and results in a higher yield than the wild-type ferredoxin-NADP+ reductase-TxtE reaction. This electrochemically driven reaction also produces two nitro-l-Trp isomers, i.e., 4-nitro-l-Trp and 5-nitro-l-Trp, which can be regulated by an electric field and electron mediators. Moreover, when more polarized mediators are used, the selectivity is recovered to 4-nitro-l-Trp. The mechanistic study suggests that the selectivity of the electron mediators is attributed to the steric hindrance and polarity of bipyridiniums. This work affords a new catalytic strategy for the regulation and control of electroenzymatic catalysis, which is expected to extend the application of electroenzymatic reactions in synthesis, energy, and sensing fields.
研究领域:
电化学
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酶学
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有机化学
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催化
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产品关联
DOI
高影响因子
Cell Reports
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2024
·
IF 7.5
Yu Chen,Yansong Fu,Yanwei Xia,Youzhi Miao,Jiahui Shao,Wei Xuan,Yunpeng Liu,Weibing Xun,Qiuyan Yan,Qirong Shen,Ruifu Zhang
Trichoderma spp. have evolved the capacity to communicate with plants by producing various secondary metabolites (SMs). Nonhormonal SMs play important roles in plant root development, while specific SMs from rhizosphere microbes and their underlying mechanisms to control plant root branching are still largely unknown. In this study, a compound, anthranilic acid (2-AA), is identified from T. guizhouense NJAU4742 to promote lateral root development. Further studies demonstrate that 2-AA positively regulates auxin signaling and transport in the canonical auxin pathway. 2-AA also partly rescues the lateral root numbers of CASP1 pro :shy2-2 , which regulates endodermal cell wall remodeling via an RBOHF -induced reactive oxygen species burst. In addition, our work reports another role for microbial 2-AA in the regulation of lateral root development, which is different from its better-known role in plant indole-3-acetic acid biosynthesis. In summary, this study identifies 2-AA from T. guizhouense NJAU4742, which plays versatile roles in regulating plant root development.
研究领域:
植物学
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分子生物学
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微生物学
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DOI
PubMed
高影响因子
ENVIRONMENTAL POLLUTION
·
2016
·
IF 4.839
Dezhen Wang,Xinru Wang,Ping Zhang,Yao Wang,Renke Zhang,Jin Yan,Zhiqiang Zhou,Wentao Zhu
Epidemiological studies indicate that exposure to persistent organic pollutants is positively associated with the prevalence of obesity. To delineate the potential role of technical-grade chlordane in obesity development, chlordane metabolism and chlordane-induced metabolic changes were investigated in mice fed high-fat diet (HFD) over a 6-week period. Gas chromatography–electron capture detector analysis showed that HFD induced more accumulation of technical chlordane in the liver, muscle and adipose tissue . The enantioselectivities of oxychlordane in selected tissues were also influenced by HFD. 1 H NMR-based liver metabolome indicated that technical chlordane can enhance the metabolic alterations induced by HFD. Compared with the low-fat diet (LFD) group, no differences were observed in the LFD + chlordane group. However, as many as 16 metabolites were significantly different between the HFD group and HFD + chlordane group. Moreover, compared to the LFD + chlordane group, the abundances of 24 metabolites significantly increased or decreased in the HFD + chlordane group. Twenty metabolites were altered in the HFD group compared to the LFD group. Tryptophan profiling suggested that both chlordane and HFD can disturb tryptophan catabolism. These interactions between technical chlordane and HFD suggest that technical chlordane is a candidate obesogen.
研究领域:
环境科学
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营养学
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毒理学
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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）