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  1. Article ; Online: Statistical optimization of bioethanol production from giant reed hydrolysate by Candida tropicalis using Taguchi design.

    Madian, Hekmat R / Hamouda, Hamed I / Hosny, Mohamed

    Journal of biotechnology

    2022  Volume 360, Page(s) 71–78

    Abstract: The economic production of bioethanol as a sustainable liquid fuel is particularly needed and attractive. Giant reed as a low-cost and renewable biomass can be utilized as a sustainable feedstock for bioethanol development. The current research focuses ... ...

    Abstract The economic production of bioethanol as a sustainable liquid fuel is particularly needed and attractive. Giant reed as a low-cost and renewable biomass can be utilized as a sustainable feedstock for bioethanol development. The current research focuses on optimizing the fermentation parameters to increase ethanol concentration while lowering production costs. In this work, the giant reed was hydrolyzed thermochemically using HCl; cellulose and hemicellulose fractions were maximally converted at optimized hydrolysis conditions (5% HCl, 30 min, and 120 °C), resulting in a high sugar concentration (≈ 55 g/L), which were fermented by Candida tropicalis Y-26 for bioethanol production (≈ 15 g/L). Taguchi design was used to optimize the fermentation parameters (temperatures, pH, incubation period, and nitrogen sources). Under optimum fermentation conditions (25 °C; 24 h.; pH 5.5; and ammonium nitrate as a nitrogen source), the ethanol concentration at flask level accomplished ≈ 21 g/L, while its scale-up to bioreactor level contributed ≈ 25 g/L (equivalent to 250 kg ethanol/ton biomass) with ≈ 67% increase than the fermentation under unoptimized conditions. Overall, these findings proved that optimizing the fermentation parameters by Taguchi design and scaling up at a bioreactor could improve bioethanol production from giant reed biomass.
    MeSH term(s) Candida tropicalis ; Ethanol
    Chemical Substances Ethanol (3K9958V90M)
    Language English
    Publishing date 2022-10-19
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 843647-2
    ISSN 1873-4863 ; 0168-1656 ; 1389-0352
    ISSN (online) 1873-4863
    ISSN 0168-1656 ; 1389-0352
    DOI 10.1016/j.jbiotec.2022.10.007
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Distinct roles of carbohydrate-binding modules in multidomain β-1,3-1,4-glucanase on polysaccharide degradation.

    Hamouda, Hamed I / Fan, Yi-Xuan / Abdalla, Mohnad / Su, Hang / Lu, Ming / Li, Fu-Li

    Applied microbiology and biotechnology

    2023  Volume 107, Issue 5-6, Page(s) 1751–1764

    Abstract: Lam16A is a novel GH16 β-1,3-1,4-lichenase isolated from the genus Caldicellulosiruptor which can utilize untreated carbohydrate components of plant cell walls. Its catalytic module has been characterized that the six carbohydrate-binding modules (CBMs) ... ...

    Abstract Lam16A is a novel GH16 β-1,3-1,4-lichenase isolated from the genus Caldicellulosiruptor which can utilize untreated carbohydrate components of plant cell walls. Its catalytic module has been characterized that the six carbohydrate-binding modules (CBMs) were queued in the C-terminus, but their roles were still unclear. Here, full-length and CBM-truncated mutants of Lam16A were purified and characterized through heterologous expression in Escherichia coli. The profiles of these proteins, including the enzyme activity, degrading efficiency, substrate-binding affinity, and thermostability, were explored. Full-length Lam16A with six CBMs showed excellent thermostability and the highest activity against barley β-glucan and laminarin with optimum pH of 6.5. The CBMs stimulated degrading ability of the catalytic module, especially against β-1,3(4)-glucan-based polysaccharides. The released products from β-1,3-1,4-glucan by Lam16A or its truncated mutants revealed an endo-type glycoside hydrolase. Lam16As exhibited strong binding affinities to the insoluble polysaccharides, especially Lam16A-1CBM. The degradation of yeast cell walls by Lam16A enzyme solution relative to the control reduced the absorbance values at OD
    MeSH term(s) Polysaccharides/metabolism ; beta-Glucans/metabolism ; Glycoside Hydrolases/metabolism ; Oligosaccharides ; Substrate Specificity
    Chemical Substances Polysaccharides ; beta-Glucans ; Glycoside Hydrolases (EC 3.2.1.-) ; Oligosaccharides
    Language English
    Publishing date 2023-02-17
    Publishing country Germany
    Document type Journal Article
    ZDB-ID 392453-1
    ISSN 1432-0614 ; 0171-1741 ; 0175-7598
    ISSN (online) 1432-0614
    ISSN 0171-1741 ; 0175-7598
    DOI 10.1007/s00253-023-12416-4
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  3. Article ; Online: Marine phospholipid nanoliposomes: A promising therapeutic approach for inflammatory bowel disease: Preparation, safety, and efficacy evaluation.

    Shabana, Samah / Hamouda, Hamed I / Hamadou, Alkassoumi Hassane / Ahmed, Busati / Chi, Zhe / Liu, Chenguang

    Colloids and surfaces. B, Biointerfaces

    2023  Volume 234, Page(s) 113702

    Abstract: Promising findings have been emerged from studies utilizing n3 polyunsaturated fatty acids (PUFA) supplementation in animal models of inflammatory bowel disease (IBD). Introduction of marine phospholipids which combine n3 PUFA with phosphatidylcholine in ...

    Abstract Promising findings have been emerged from studies utilizing n3 polyunsaturated fatty acids (PUFA) supplementation in animal models of inflammatory bowel disease (IBD). Introduction of marine phospholipids which combine n3 PUFA with phosphatidylcholine in a nanoliposome formulation offers enhanced pharmacological efficacy due to physical stability, improved bioavailability, and specific targeting to inflamed colitis tissues. In the present study, a marine phospholipid-based nanoliposome formulation was developed and optimized, resulting in nanovesicles of approximately 107.7 ± 1.3 nm in size, 0.18 ± 0.01 PDI, and - 32.03 ± 3.16 mV ZP. The nanoliposomes exhibited spherical vesicles with stable properties upon incubation at SGF as shown by the TEM, DLS, and turbidity measurements over 3 h. MPL nanoliposomes were cytocompatible until the concentration of 500 µg/mL as per MTT assay and taken by macrophages through macropinocytosis and caveolae pathways, and demonstrated significant inhibitory activity against reactive oxygen species (ROS) in LPS-stimulated macrophages. They were also shown to be blood-compatible and safe for administration in healthy mice. In a colitis mouse model, the nanoliposomes displayed preferential distribution in the inflamed gut, delaying the onset of colitis when administered prophylactically. These findings highlight the potential of marine phospholipid nanoliposomes as a promising therapeutic approach for managing inflammatory bowel disease.
    MeSH term(s) Animals ; Mice ; Phospholipids ; Inflammatory Bowel Diseases/drug therapy ; Colitis/chemically induced ; Colitis/drug therapy ; Fatty Acids, Omega-3 ; Phosphatidylcholines ; Liposomes
    Chemical Substances Phospholipids ; Fatty Acids, Omega-3 ; Phosphatidylcholines ; Liposomes
    Language English
    Publishing date 2023-12-07
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 1500523-9
    ISSN 1873-4367 ; 0927-7765
    ISSN (online) 1873-4367
    ISSN 0927-7765
    DOI 10.1016/j.colsurfb.2023.113702
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  4. Article ; Online: Distinct roles of carbohydrate-binding modules in multidomain β-1,3–1,4-glucanase on polysaccharide degradation

    Hamouda, Hamed I. / Fan, Yi-Xuan / Abdalla, Mohnad / Su, Hang / Lu, Ming / Li, Fu-Li

    Appl Microbiol Biotechnol. 2023 Mar., v. 107, no. 5-6 p.1751-1764

    2023  

    Abstract: Lam16A is a novel GH16 β-1,3–1,4-lichenase isolated from the genus Caldicellulosiruptor which can utilize untreated carbohydrate components of plant cell walls. Its catalytic module has been characterized that the six carbohydrate-binding modules (CBMs) ... ...

    Abstract Lam16A is a novel GH16 β-1,3–1,4-lichenase isolated from the genus Caldicellulosiruptor which can utilize untreated carbohydrate components of plant cell walls. Its catalytic module has been characterized that the six carbohydrate-binding modules (CBMs) were queued in the C-terminus, but their roles were still unclear. Here, full-length and CBM-truncated mutants of Lam16A were purified and characterized through heterologous expression in Escherichia coli. The profiles of these proteins, including the enzyme activity, degrading efficiency, substrate-binding affinity, and thermostability, were explored. Full-length Lam16A with six CBMs showed excellent thermostability and the highest activity against barley β-glucan and laminarin with optimum pH of 6.5. The CBMs stimulated degrading ability of the catalytic module, especially against β-1,3(4)-glucan-based polysaccharides. The released products from β-1,3–1,4-glucan by Lam16A or its truncated mutants revealed an endo-type glycoside hydrolase. Lam16As exhibited strong binding affinities to the insoluble polysaccharides, especially Lam16A-1CBM. The degradation of yeast cell walls by Lam16A enzyme solution relative to the control reduced the absorbance values at OD₈₀₀ by ~ 85% ± 1.2, enabling the release of up to ~ 0.057 ± 0.0039 µg/mL of the cytoplasmic protein into the supernatant, lowering the viability of the cells by ~ 70.3% ± 6.9, thus causing significant damage in the cell wall structure. Taken together, CBMs could influence the substrate specificity, thermal stability, and binding affinity of β-1,3–1,4-glucanase. These results demonstrate the great potential of these enzymes to promote the bioavailability of β-1,3-glucan oligosaccharides for health benefits. KEY POINTS: • Carbohydrate-binding modules strongly influenced the enzyme activity and binding affinity, and further impacted glycoside hydrolase activity. • Lam16A enzymes have sufficient ability to hydrolyze β-1,3–1,4-glucan-based polysaccharides. • Lam16As provide a powerful tool to promote the bioavailability of β-1,3-glucan oligosaccharides.
    Keywords Caldicellulosiruptor ; Escherichia coli ; absorbance ; barley ; bioavailability ; carbohydrate binding ; cell walls ; enzyme activity ; glycosides ; heterologous gene expression ; hydrolases ; oligosaccharides ; pH ; polysaccharides ; substrate specificity ; thermal stability ; viability ; yeasts
    Language English
    Dates of publication 2023-03
    Size p. 1751-1764.
    Publishing place Springer Berlin Heidelberg
    Document type Article ; Online
    ZDB-ID 392453-1
    ISSN 1432-0614 ; 0171-1741 ; 0175-7598
    ISSN (online) 1432-0614
    ISSN 0171-1741 ; 0175-7598
    DOI 10.1007/s00253-023-12416-4
    Database NAL-Catalogue (AGRICOLA)

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  5. Article: Biochemical Characterization of a Cold-Adapted λ-Carrageenase OUC-CglA from Maribacter vaceletii: An Efficient Tool for λ-Carrageenan Degradation

    Lu, Zewei / Jiang, Hong / Hamouda, Hamed I. / Wang, Tao / Dong, Yueyang / Mao, Xiangzhao

    Journal of agricultural and food chemistry. 2022 Sept. 16, v. 70, no. 38

    2022  

    Abstract: λ-Carrageenase with high activity is an effective and environmentally friendly tool enzyme for the preparation of λ-carrageenan oligosaccharides with various biological activities. Herein, a novel GH150 (glycoside hydrolases family 150) λ-carrageenase ... ...

    Abstract λ-Carrageenase with high activity is an effective and environmentally friendly tool enzyme for the preparation of λ-carrageenan oligosaccharides with various biological activities. Herein, a novel GH150 (glycoside hydrolases family 150) λ-carrageenase OUC-CglA from Maribacter vaceletii was heterologously expressed, purified, and characterized. The recombinant OUC-CglA performs strict selectivity toward λ-carrageenan with a specific activity of 418.7 U/mg under its optimal reaction conditions of 20 °C and pH 7.0. Additionally, OUC-CglA is a typical cold-adapted λ-carrageenase because it unfolds 90% and 63% of its maximum activity at 15 and 10 °C, respectively. The hydrolysis process suggests that OUC-CglA is an endotype λ-carrageenase with the final products consisting of λ-neocarrabiose, λ-neocarratetraose, λ-neocarrahexaose, and other long-chain λ-neocarrageenan oligosaccharides. As a result, high activity, cold-adaptation, and principal products of OUC-CglA make it a potential biocatalyst for the effective preparation of λ-carrageenan oligosaccharides.
    Keywords biocatalysts ; food chemistry ; glycosidases ; hydrolysis ; oligosaccharides ; pH
    Language English
    Dates of publication 2022-0916
    Size p. 12135-12142.
    Publishing place American Chemical Society
    Document type Article
    ZDB-ID 241619-0
    ISSN 1520-5118 ; 0021-8561
    ISSN (online) 1520-5118
    ISSN 0021-8561
    DOI 10.1021/acs.jafc.2c05544
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  6. Article ; Online: Biochemical Characterization of a Cold-Adapted λ-Carrageenase OUC-CglA from

    Lu, Zewei / Jiang, Hong / Hamouda, Hamed I / Wang, Tao / Dong, Yueyang / Mao, Xiangzhao

    Journal of agricultural and food chemistry

    2022  Volume 70, Issue 38, Page(s) 12135–12142

    Abstract: λ-Carrageenase with high activity is an effective and environmentally friendly tool enzyme for the preparation of λ-carrageenan oligosaccharides with various biological activities. Herein, a novel GH150 (glycoside hydrolases family 150) λ-carrageenase ... ...

    Abstract λ-Carrageenase with high activity is an effective and environmentally friendly tool enzyme for the preparation of λ-carrageenan oligosaccharides with various biological activities. Herein, a novel GH150 (glycoside hydrolases family 150) λ-carrageenase OUC-CglA from
    MeSH term(s) Bacterial Proteins/metabolism ; Carrageenan/chemistry ; Flavobacteriaceae ; Glycoside Hydrolases/chemistry ; Oligosaccharides/chemistry
    Chemical Substances Bacterial Proteins ; Oligosaccharides ; Carrageenan (9000-07-1) ; Glycoside Hydrolases (EC 3.2.1.-)
    Language English
    Publishing date 2022-09-16
    Publishing country United States
    Document type Journal Article
    ZDB-ID 241619-0
    ISSN 1520-5118 ; 0021-8561
    ISSN (online) 1520-5118
    ISSN 0021-8561
    DOI 10.1021/acs.jafc.2c05544
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  7. Article ; Online: Multifunctional nanoparticles based on marine polysaccharides for apremilast delivery to inflammatory macrophages: Preparation, targeting ability, and uptake mechanism.

    Shabana, Samah / Hamouda, Hamed I / Abdalla, Mohnad / Sharaf, Mohamed / Chi, Zhe / Liu, Chenguang

    International journal of biological macromolecules

    2022  Volume 222, Issue Pt B, Page(s) 1709–1722

    Abstract: Hydrophobic drug encapsulation inside targeted nanoparticles can enhance accumulation in inflamed sites, limit toxicity to healthy tissue, and improve pharmacokinetics compared to free drug dosing. This study reports a functionalized marine ... ...

    Abstract Hydrophobic drug encapsulation inside targeted nanoparticles can enhance accumulation in inflamed sites, limit toxicity to healthy tissue, and improve pharmacokinetics compared to free drug dosing. This study reports a functionalized marine polysaccharide nanoparticle with a controlled release, targeting abilities, and in-situ imaging properties. Carbon dots functionalized Enteromorpha polysaccharide/Mannose/Methionine functionalized Chitosan (CDs.EP/Man/Meth.Cs) NPs could deliver apremilast to inflammatory macrophages and Caco-2 intestinal cells as an in vitro model for application in oral drug delivery to cure IBD. The nanoparticles were simply a polyelectrolyte complex between cationic functionalized chitosan and anionic polysaccharide of Enteromorpha prolifera. Functionalized polysaccharides and the prepared NPs were well characterized. The functionalized nanoparticles could overcome the limitation of poor drug bioavailability and showed a high loading capacity of (45 %) with a controlled release of about (74.5 %). Confocal laser scanning imaging showed higher cellular uptake of the modified nanoparticles than that of the unmodified nanoparticles in LPS-activated RAW 264.7 macrophages and Caco-2 cells. The effect of functionalization on the cellular uptake targetability was assessed using spectrofluorometric measurements after mannose competition. Anti-inflammatory activity of apremilast-loaded NPs is more elevated than the free drug. These results suggest the feasibility of using functionalized EP/Cs nanoparticles in IBD oral drug delivery.
    MeSH term(s) Humans ; Chitosan/chemistry ; Drug Carriers/chemistry ; Multifunctional Nanoparticles ; Caco-2 Cells ; Mannose ; Delayed-Action Preparations ; Drug Delivery Systems/methods ; Nanoparticles/chemistry ; Polysaccharides/pharmacology ; Polysaccharides/chemistry ; Macrophages ; Inflammatory Bowel Diseases
    Chemical Substances Chitosan (9012-76-4) ; Drug Carriers ; apremilast (UP7QBP99PN) ; Mannose (PHA4727WTP) ; Delayed-Action Preparations ; Polysaccharides
    Language English
    Publishing date 2022-09-28
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 282732-3
    ISSN 1879-0003 ; 0141-8130
    ISSN (online) 1879-0003
    ISSN 0141-8130
    DOI 10.1016/j.ijbiomac.2022.09.225
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  8. Article ; Online: Catalytic properties characterization and degradation mode elucidation of a polyG-specific alginate lyase OUC-FaAly7.

    Chen, Yimiao / Ci, Fangfang / Jiang, Hong / Meng, Di / Hamouda, Hamed I / Liu, Chunhui / Quan, Yongyi / Chen, Suxue / Bai, Xinxue / Zhang, Zhaohui / Gao, Xin / Balah, Mohamed A / Mao, Xiangzhao

    Carbohydrate polymers

    2024  Volume 333, Page(s) 121929

    Abstract: Polymerized guluronates (polyG)-specific alginate lyase with lower polymerized mannuronates (polyM)-degrading activity, superior stability, and clear action mode is a powerful biotechnology tool for the preparation of AOSs rich in M blocks. In this study, ...

    Abstract Polymerized guluronates (polyG)-specific alginate lyase with lower polymerized mannuronates (polyM)-degrading activity, superior stability, and clear action mode is a powerful biotechnology tool for the preparation of AOSs rich in M blocks. In this study, we expressed and characterized a polyG-specific alginate lyase OUC-FaAly7 from Formosa agariphila KMM3901. OUC-FaAly7 belonging to polysaccharide lyase (PL) family 7 had highest activity (2743.7 ± 20.3 U/μmol) at 45 °C and pH 6.0. Surprisingly, its specific activity against polyG reached 8560.2 ± 76.7 U/μmol, whereas its polyM-degrading activity was nearly 0 within 10 min reaction. Suggesting that OUC-FaAly7 was a strict polyG-specific alginate lyase. Importantly, OUC-FaAly7 showed a wide range of temperature adaptations and remarkable temperature and pH stability. Its relative activity between 20 °C and 45 °C reached >90 % of the maximum activity. The minimum identifiable substrate of OUC-FaAly7 was guluronate tetrasaccharide (G4). Action process and mode showed that it was a novel alginate lyase digesting guluronate hexaose (G6), guluronate heptaose (G7), and polymerized guluronates, with the preferential generation of unsaturated guluronate pentasaccharide (UG5), although which could be further degraded into unsaturated guluronate disaccharide (UG3) and trisaccharide (UG2). This study contributes to illustrating the catalytic properties, substrate recognition, and action mode of novel polyG-specific alginate lyases.
    MeSH term(s) Substrate Specificity ; Oligosaccharides/metabolism ; Disaccharides/metabolism ; Polysaccharide-Lyases/metabolism ; Alginates/metabolism ; Hydrogen-Ion Concentration ; Bacterial Proteins/chemistry
    Chemical Substances poly(beta-D-mannuronate) lyase (EC 4.2.2.3) ; Oligosaccharides ; Disaccharides ; Polysaccharide-Lyases (EC 4.2.2.-) ; Alginates ; Bacterial Proteins
    Language English
    Publishing date 2024-02-15
    Publishing country England
    Document type Journal Article
    ZDB-ID 1501516-6
    ISSN 1879-1344 ; 0144-8617
    ISSN (online) 1879-1344
    ISSN 0144-8617
    DOI 10.1016/j.carbpol.2024.121929
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  9. Article: Biochemical Characterization of a Novel Myrosinase Rmyr from Rahnella inusitata for High-Level Preparation of Sulforaphene and Sulforaphane

    Wang, Lili / Jiang, Hong / Qiu, Yanjun / Dong, Yueyang / Hamouda, Hamed I. / Balah, Mohamed A. / Mao, Xiangzhao

    Journal of agricultural and food chemistry. 2022 Feb. 03, v. 70, no. 7

    2022  

    Abstract: Myrosinase is a biotechnological tool for the preparation of sulforaphane and sulforaphene with a variety of excellent biological activities. In this study, a gene encoding the novel glycoside hydrolase family 3 (GH3) myrosinase Rmyr from Rahnella ... ...

    Abstract Myrosinase is a biotechnological tool for the preparation of sulforaphane and sulforaphene with a variety of excellent biological activities. In this study, a gene encoding the novel glycoside hydrolase family 3 (GH3) myrosinase Rmyr from Rahnella inusitata was heterologously expressed in Escherichia coli BL21 (DE3). The purified Rmyr shows the highest activity at 40 °C and pH 7.0; meanwhile, its half-life at 30 °C reaches 12 days, indicating its excellent stability. Its sinigrin-, glucoraphenin-, and glucoraphanin-hydrolyzing activities were 12.73, 4.81, and 6.99 U/mg, respectively. Rmyr could efficiently degrade the radish seed-derived glucoraphenin and the broccoli seed-derived glucoraphanin into sulforaphene and sulforaphane within 10 min with the highest yields of 5.07 mg/g radish seeds and 9.56 mg/g broccoli seeds, respectively. The highest conversion efficiencies of sulforaphane from glucoraphanin and sulforaphene from glucoraphenin reached up to 92.48 and 97.84%, respectively. Therefore, Rmyr is a promising and potent biocatalyst for efficient and large-scale preparation of sulforaphane and sulforaphene.
    Keywords Escherichia coli ; Rahnella ; biocatalysts ; broccoli ; food chemistry ; genes ; glucoraphanin ; half life ; pH ; radishes ; sulforaphane ; thioglucosidase
    Language English
    Dates of publication 2022-0203
    Size p. 2303-2311.
    Publishing place American Chemical Society
    Document type Article
    ZDB-ID 241619-0
    ISSN 1520-5118 ; 0021-8561
    ISSN (online) 1520-5118
    ISSN 0021-8561
    DOI 10.1021/acs.jafc.1c07646
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  10. Article: Expression and Biochemical Characterization of a Novel Fucoidanase from

    Qiu, Yanjun / Jiang, Hong / Dong, Yueyang / Wang, Yongzhen / Hamouda, Hamed I / Balah, Mohamed A / Mao, Xiangzhao

    Foods (Basel, Switzerland)

    2022  Volume 11, Issue 7

    Abstract: Fucoidan is one of the main polysaccharides of brown algae and echinoderm, which has nutritional and pharmacological functions. Due to the low molecular weight and exposure of more sulfate groups, oligo-fucoidan or fucoidan oligosaccharides have ... ...

    Abstract Fucoidan is one of the main polysaccharides of brown algae and echinoderm, which has nutritional and pharmacological functions. Due to the low molecular weight and exposure of more sulfate groups, oligo-fucoidan or fucoidan oligosaccharides have potential for broader applications. In this research, a novel endo-α-1,4-L-fucoidanase OUC-FaFcn1 which can degrade fucoidan into oligo-fucoidan was discovered from the fucoidan-digesting strain
    Language English
    Publishing date 2022-04-01
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2704223-6
    ISSN 2304-8158
    ISSN 2304-8158
    DOI 10.3390/foods11071025
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