Publications by authors named "Chen-guang Liu"

77 Publications

Omics analysis reveals mechanism underlying metabolic oscillation during continuous very-high-gravity ethanol fermentation by Saccharomyces cerevisiae.

Biotechnol Bioeng 2021 08 13;118(8):2990-3001. Epub 2021 May 13.

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

During continuous very-high-gravity (VHG) ethanol fermentation with Saccharomyces cerevisiae, the process exhibits sustained oscillation in residual glucose, ethanol, and biomass, raising a question: how do yeast cells respond to this phenomenon? In this study, the oscillatory behavior of yeast cells was characterized through transcriptome and metabolome analysis for one complete oscillatory period. By analyzing the accumulation of 26 intracellular metabolites and the expression of 90 genes related to central carbon metabolism and stress response, we confirmed that the process oscillation was attributed to intracellular metabolic oscillation with phase difference, and the expression of HXK1, HXT1,2,4, and PFK1 was significantly different from other genes in the Embden-Meyerhof-Parnas pathway, indicating that glucose transport and phosphorylation could be key nodes for regulating the intracellular metabolism under oscillatory conditions. Moreover, the expression of stress response genes was triggered and affected predominately by ethanol inhibition in yeast cells. This progress not only contributes to the understanding of mechanisms underlying the process oscillation observed for continuous VHG ethanol fermentation, but also provides insights for understanding unsteady state that might develop in other continuous fermentation processes operated under VHG conditions to increase product titers for robust production.
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http://dx.doi.org/10.1002/bit.27809DOI Listing
August 2021

Upgrading of Light Bio-oil from Solvothermolysis Liquefaction of an Oil Palm Empty Fruit Bunch in Glycerol by Catalytic Hydrodeoxygenation Using NiMo/AlO or CoMo/AlO Catalysts.

ACS Omega 2021 Feb 21;6(4):2999-3016. Epub 2021 Jan 21.

Department of Chemical Engineering, Faculty of Engineering, Mahidol University, 25/25 Putthamonthon 4 Road, Salaya, Phutthamonthon, Nakhon Pathom 73170, Thailand.

Hydrodeoxygenation (HDO) of bio-oil derived from liquefaction of a palm empty fruit bunch (EFB) in glycerol was investigated. To enhance the heating value and reduce the oxygen content of upgraded bio-oil, hydrodeoxygenation of light bio-oil over Ni- and Co-based catalysts on an AlO support was performed in a rotating-bed reactor. Two consecutive steps were conducted to produce bio-oil from EFB including (1) microwave-assisted wet torrefaction of EFB and (2) solvothermolysis liquefaction of treated EFB in a NaCO/glycerol system. The HDO of as-prepared bio-oil was subsequently performed in a unique design reactor possessing a rotating catalyst bed for efficient interaction of a catalyst with bio-oil and facile separation of the catalyst from upgraded bio-oil after the reaction. The reaction was carried out in the presence of each mono- or bimetallic catalyst, namely, Co/AlO, Ni/AlO, NiMo/AlO, and CoMo/AlO, packed in the rotating-mesh host with a rotation speed of 250 rpm and kept at 300 and 350 °C, 2 MPa hydrogen for 1 h. From the results, the qualities of upgraded bio-oil were substantially improved for all catalysts tested in terms of oxygen reduction and increased high heating value (HHV). Particularly, the NiMo/AlO catalyst exhibited the most promising catalyst, providing favorable bio-oil yield and HHV. Remarkably greater energy ratios and carbon recovery together with high H/O, C/O, and H/C ratios were additionally achieved from the NiMo/AlO catalyst compared with other catalysts. Cyclopentanone and cyclopentene were the main olefins found in hydrodeoxygenated bio-oil derived from liquefied EFB. It was observed that cyclopentene was first generated and subsequently converted to cyclopentanone under the hydrogenation reaction. These compounds can be further used as a building block in the synthesis of jet-fuel range cycloalkanes.
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http://dx.doi.org/10.1021/acsomega.0c05387DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7860089PMC
February 2021

Low-temperature extrusion-based 3D printing of icariin-laden scaffolds for osteogenesis enrichment.

Regen Ther 2021 Mar 21;16:53-62. Epub 2021 Jan 21.

Institute of Biomaterials and Tissue Engineering, Huaqiao University, Xiamen, Fujian, 361021, PR China.

Despite the accessibility to porous architectures through various biofabrication approaches for tissue engineering, incorporating various active growth regulators within their matrices that act as biochemical cues is also an essential attribute for effective tissue growth. To address these facts, icariin (ICA)-encapsulated polymeric scaffolds are fabricated using a low-temperature extrusion-based three-dimensional (3D) printing technology for efficiently promoting osteogenesis. This approach not only resulted in the generation of porous architectures but also substantially maintained the bio-efficacy of the encapsulated ICA. Moreover, these composite scaffolds based on poly(ε-caprolactone) (PCL) and tricalcium phosphate (β-TCP) encapsulated with ICA (ITP scaffolds) are systematically characterized using various techniques before and after printing. Furthermore, various investigations relevant to biodegradability, biocompatibility, ICA release, and osteogenic ability of the ITP scaffolds are explored. The intact physiochemical properties of the materials, sustained release of ICA from the scaffolds, and high biosafety at various levels ranging from cellular to animal efficiently promoted the proliferation of mouse bone marrow mesenchymal stem cells (BMSCs) and their differentiation to osteoblasts. Together, the utilization of low-temperature extrusion approach provides a convenient and eco-friendly means of fabricating highly porous 3D architectures that supply the required growth regulators in their active form for tissue regeneration.
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http://dx.doi.org/10.1016/j.reth.2021.01.001DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7820910PMC
March 2021

Gut microbiota homeostasis restoration may become a novel therapy for breast cancer.

Invest New Drugs 2021 Jun 17;39(3):871-878. Epub 2021 Jan 17.

Department of Infectious disease, Jingzhou Central Hospital, The Second Clinical Medical College, Yangtze University, No. 60 Jingzhong Road, Jingzhou District, Jingzhou, 434020, Hubei Province, China.

Breast cancer is the most diagnosed cancer in women. It significantly impairs a patient's physical and mental health. Gut microbiota comprise the bacteria residing in a host's gastrointestinal tract. Through studies over the last decade, we now know that alterations in the composition of the gut microbiome are associated with protection against colonization by pathogens and other diseases, such as diabetes and cancer. This review focuses on how gut microbiota can affect breast cancer development through estrogen activity and discusses the types of bacteria that may be involved in the onset and the progression of breast cancer. We also describe potential therapies to curtail the risk of breast cancer by restoring gut microbiota homeostasis and reducing systemic estrogen levels. This review will further explore the relationship between intestinal microbes and breast cancer and propose a method to treat breast cancer by improving intestinal microbes. We aimed at discovering new methods to prevent or treat BC by changing intestinal microorganisms.
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http://dx.doi.org/10.1007/s10637-021-01063-zDOI Listing
June 2021

Chitosan-based nanoparticles as delivery-carrier for promising antimicrobial glycolipid biosurfactant to improve the eradication rate of biofilm.

J Biomater Sci Polym Ed 2021 04 11;32(6):813-832. Epub 2021 Feb 11.

College of Marine Life Science, Ocean University Of China, Qingdao, P.R. China.

Driven by the need to find alternatives to control infections, this work describes the development of chitosan-PMLA nanoparticulate systems as carriers for antimicrobial glycolipid. By using a simple ionic gelation method stable nanoparticle was obtained showing an encapsulation efficiency of 73.1 ± 1.3% and an average size of 217.0 ± 15.6 nm for rhamnolipids chitosan-PMLA nanoparticles (RL-CS-NPs). Glycolipid incorporation and particle size were correspondingly corroborated by FT-IR and TEM analysis. Rhamnolipids chitosan nanoparticles (RL-CS-NPs) presented the highest antimicrobial effect towards (ATCC 26695) exhibiting a minimal inhibitory concentration of 132 µg/mL and a biofilm inhibition ability of 99%. Additionally, RL-CS-NPs did not interfere with human fibroblasts viability and proliferation under the tested conditions. The results revealed that the RL-CS-NPs were able to inhibit bacterial growth showing adequate cytocompatibility and might become, after additional studies, a valuable approach to fight biofilm related-infections.
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http://dx.doi.org/10.1080/09205063.2020.1870323DOI Listing
April 2021

Elucidating the pyrolysis reaction mechanism of Calotropis procera and analysis of pyrolysis products to evaluate its potential for bioenergy and chemicals.

Bioresour Technol 2021 Feb 15;322:124545. Epub 2020 Dec 15.

Government College University Faisalabad, Faisalabad 38000, Pakistan. Electronic address:

The present study was focused on evaluating the bioenergy potential of waste biomass of desert plant Calotropis procera. The biomass was pyrolyzed at four heating rates including 10 °Cmin, 20 °Cmin, 40 °Cmin, and 80 °Cmin. The pyrolysis reaction kinetics and thermodynamics parameters were assessed using isoconversional models namely Kissenger-Akahira-Sunose, Flynn-Wall-Ozawa, and Starink. Major pyrolysis reaction occurred between 200 and 450 °C at the conversion points (α) ranging from 0.2 to 0.6 while their corresponding reaction parameters including activation energy, enthalpy change, Gibb's free energy and pre-exponential factors were ranged from 165 to 207 kJ mol, 169-200 kJ mol, 90-42 kJ mol, and 10-10 s, respectively. The narrow range of pre-exponential factors indicated a uniform pyrolysis, while lower differences between enthalpy change and activation energies indicated that reactions were thermodynamically favorable. The evolved gases were dominated by propanoic acid, 3-hydroxy-, hydrazide, hydrazinecarboxamide and carbohydrazide followed by amines/amides, alcohols, acids, aldehydes/ketones, and esters.
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http://dx.doi.org/10.1016/j.biortech.2020.124545DOI Listing
February 2021

Macrophages-targeting mannosylated nanoparticles based on inulin for the treatment of inflammatory bowel disease (IBD).

Int J Biol Macromol 2021 Feb 16;169:206-215. Epub 2020 Dec 16.

College of Marine Life Science, Ocean University Of China, No.5 Yushan Road, Qingdao 266003, Shandong, China. Electronic address:

In the present experimental series, we have developed a novel nanocomposite to target activated macrophages in the colon with real time imaging and therapeutic capabilities. This binary nanocomposite was formed by the covalent conjugation of mannosylated NPs (Man-NPs) with carbon dots (CDs). Man-NPs were prepared using a self-assembly method based on mannosylated decamethylenediamine-grafted carboxymethyl inulin amphiphilic acid. While, the CDs were synthesized using a simple bottom-up process using citric acid monohydrate and diethylenetriamine, which were tightly bonded to the Man-NPs surface by carbodimide coupling. The resulting nanocomposite had a uniform size of 241.3 nm with a negative charge and a high drug casing density of 25.54 wt% and blue self-fluorescence were emitted. Whereas, in vitro observation of cellular uptake indicated the greater nanocomposite uptake in inflamed macrophage as compared to the untreated macrophage and mannose receptor-negative cell lines, 4T1 respectively. However, in vivo bio distribution exhibited a large number (60%) of CDs/Man-NPs nanocomposite accumulated in the inflamed colon of colitis mice. It should be noted that the novel nanocomposite, as macrophage-targeted drug delivery, could have promise for the treatment of inflammatory bowel disease (IBD).
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http://dx.doi.org/10.1016/j.ijbiomac.2020.12.094DOI Listing
February 2021

Measurement of Cellulase and Xylanase Activities in Trichoderma reesei.

Methods Mol Biol 2021 ;2234:135-146

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

The microbial cellulase system is responsible for the generation of glucose from cellulose. Cellulases are comprised of at least three major groups of enzymes, namely endoglucanases, exoglucanases, and β-glucosidases. On the other hand, xylanases function in the degradation of hemicellulose and work synergistically with cellulases for the degradation of lignocellulosic biomass. Here, we describe the most commonly used methods for the activity measurement of cellulases and xylanases.
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http://dx.doi.org/10.1007/978-1-0716-1048-0_12DOI Listing
March 2021

Biodegradable Quantum Composites for Synergistic Photothermal Therapy and Copper-Enhanced Chemotherapy.

ACS Appl Mater Interfaces 2020 Oct 7;12(42):47289-47298. Epub 2020 Oct 7.

Institute for Translational Medicine, School of Basic Medical Sciences, Fujian Medical University, Fuzhou 350108, P. R. China.

In recent times, the combination therapy has garnered enormous interest owing to its great potential in clinical research. It has been reported that disulfiram, a clinical antialcoholism drug, could be degraded to diethyldithiocarbamate (DDTC) and subsequently result in the copper-DDTC complex (Cu(DDTC)) toward ablating cancer cells. In addition, the ultrasmall copper sulfide nanodots (CuS NDs) have shown great potential in cancer treatment because of their excellent photothermal and photodynamic therapeutic efficiencies. Herein, by taking advantage of the interactions between CuS and DDTC, a new multifunctional nanoplatform based on DDTC-loaded CuS (CuS-DDTC) NDs is successfully fabricated, leading to the achievement of the synergistic effect of photothermal and copper enhanced chemotherapy. All experimental results verified promising synergistic therapeutic effects. Moreover, biocompatibility and metabolism experiments displayed that the CuS-DDTC NDs could be quickly excreted from the body with no apparent toxicity signs. Together, our findings indicated the superior synergistic therapeutic effect of photothermal and copper-enhanced chemotherapy, providing a promising anticancer strategy based on the CuS-DDTC NDs drug delivery system.
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http://dx.doi.org/10.1021/acsami.0c14636DOI Listing
October 2020

Near-Infrared-Activated Lysosome Pathway Death Induced by ROS Generated from Layered Double Hydroxide-Copper Sulfide Nanocomposites.

ACS Appl Mater Interfaces 2020 Sep 27;12(36):40673-40683. Epub 2020 Aug 27.

Institute of Biomaterials and Tissue Engineering, Huaqiao University, Xiamen 361021, P. R. China.

The overdeveloped lysosomes in cancer cells are gaining increasing attention toward more precise and effective organelle-targeted cancer therapy. It is suggested that rod/plate-like nanomaterials with an appropriate size exhibited a greater quantity and longer-term lysosomal enrichment, as the shape plays a notable role in the nanomaterial transmembrane process and subcellular behaviors. Herein, a biodegradable platform based on layered double hydroxide-copper sulfide nanocomposites (LDH-CuS NCs) is successfully prepared via in situ growth of CuS nanodots on LDH nanoplates. The as-prepared LDH-CuS NCs exhibited not only high photothermal conversion and near-infrared (NIR)-induced chemodynamic and photodynamic therapeutic efficacies, but also could achieve real-time in vivo photoacoustic imaging (PAI) of the entire tumor. LDH-CuS NCs accumulated in lysosomes would then generate extensive subcellular reactive oxygen species (ROS) in situ, leading to lysosomal membrane permeabilization (LMP) pathway-associated cell death both in vitro and in vivo.
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http://dx.doi.org/10.1021/acsami.0c11739DOI Listing
September 2020

Preparation, characterization, and drug release behavior of thiolated alginate nanoparticles loaded budesonide as a potential drug delivery system toward inflammatory bowel diseases.

J Biomater Sci Polym Ed 2020 12 3;31(18):2299-2317. Epub 2020 Sep 3.

College of Marine Life Sciences, Ocean University of China, Qingdao, P.R. China.

For site-specific drug delivery in inflammatory bowel disease, reducible sodium alginate nanoparticles cross-linked with disulfide linkage were developed. Nanoparticles were synthesized in deionized water through self-assembly of amphiphilic thiolated sodium alginate Alg-Cys and subsequently produced cross-linking of disulfide bonds. TEM image showed a spherical core-shell configuration with a size of about 430 nm for the nanoparticles. Dynamic light scattering (DLS) showed high stability, narrow size distribution, and pH-dependent swelling transition for the nanoparticles. Cytotoxicity study showed that there was no evident cell inhibition among the nanoparticles. Also, the size of the nanoparticles increased in 10 mM glutathione (GSH) solution due to the cleavage of disulfides within their network structures. Compared to that in GSH-free buffer, there was a remarkable increase in drug release in pH 7.4 buffer with GSH from drug-loaded nanoparticles, indicating that the nanoparticles could be used for colon-specific drug delivery.
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http://dx.doi.org/10.1080/09205063.2020.1803034DOI Listing
December 2020

Global Reprogramming of Gene Transcription in by Overexpressing an Artificial Transcription Factor for Improved Cellulase Production and Identification of Ypr1 as an Associated Regulator.

Front Bioeng Biotechnol 2020 3;8:649. Epub 2020 Jul 3.

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

Synthetic biology studies on filamentous fungi are providing unprecedented opportunities for optimizing this important category of microbial cell factory. Artificial transcription factor can be designed and used to offer novel modes of regulation on gene transcription network. is commonly used for cellulase production. In our previous studies, a plasmid library harboring genes encoding artificial zinc finger proteins (AZFPs) was constructed for engineering , and the mutant strains with improved cellulase production were selected. However, the underlying mechanism by which AZFP function remain unclear. In this study, a Rut-C30 mutant strain U5 bearing an AZFP named as AZFP-U5 was focused, which secretes high level protein and shows significantly improved cellulase and xylanase production comparing with its parental strain. In addition, enhanced sugar release was achieved from lignocellulosic biomass using the crude cellulase from U5. Comparative transcriptome analysis was further performed, which showed reprogramming of global gene transcription and elevated transcription of genes encoding glycoside hydrolases by overexpressing . Furthermore, 15 candidate regulatory genes which showed remarkable higher transcription levels by insertion were overexpressed in Rut-C30 to examine their effects on cellulase biosynthesis. Among these genes, () and showed stimulating effects on filter paper activity (FPase), but deletion of these two genes did not affect cellulase activity. In addition, increased yellow pigment production in Rut-C30 by overexpression of gene was observed, and changes of cellulase gene transcription were revealed in the deletion mutant, suggesting possible interaction between pigment production and cellulase gene transcription. The results in this study revealed novel aspects in regulation of cellulase gene expression by the artificial regulators. In addition, the candidate genes and processes identified in the transcriptome data can be further explored for synthetic biology design and metabolic engineering of to enhance cellulase production.
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http://dx.doi.org/10.3389/fbioe.2020.00649DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7351519PMC
July 2020

Engineering the Effector Domain of the Artificial Transcription Factor to Improve Cellulase Production by .

Front Bioeng Biotechnol 2020 25;8:675. Epub 2020 Jun 25.

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

Filamentous fungal strains of have been widely used for cellulase production, and great effort has been devoted to enhancing their cellulase titers for the economic biorefinery of lignocellulosic biomass. In our previous studies, artificial zinc finger proteins (AZFPs) with the Gal4 effector domain were used to enhance cellulase biosynthesis in , and it is of great interest to modify the AZFPs to further improve cellulase production. In this study, the endogenous activation domain from the transcription activator Xyr1 was used to replace the activation domain of Gal4 of the AZFP to explore impact on cellulase production. The cellulase producer TU-6 was used as a host strain, and the engineered strains containing the Xyr1 and the Gal4 activation domains were named as QS2 and QS1, respectively. Compared to QS1, activities of filter paper and endoglucanases in crude cellulase produced by QS2 increased 24.6 and 50.4%, respectively. Real-time qPCR analysis also revealed significant up-regulation of major genes encoding cellulase in QS2. Furthermore, the biomass hydrolytic performance of the cellulase was evaluated, and 83.8 and 97.9% more glucose was released during the hydrolysis of pretreated corn stover using crude enzyme produced by QS2, when compared to the hydrolysis with cellulase produced by QS1 and the parent strain TU-6. As a result, we proved that the effector domain in the AZFPs can be optimized to construct more effective artificial transcription factors for engineering to improve its cellulase production.
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http://dx.doi.org/10.3389/fbioe.2020.00675DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7330100PMC
June 2020

Intracellular Redox Perturbation in Improved Furfural Tolerance and Enhanced Cellulosic Bioethanol Production.

Front Bioeng Biotechnol 2020 23;8:615. Epub 2020 Jun 23.

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

Furfural is a major toxic byproduct found in the hydrolysate of lignocellulosic biomass, which adversely interferes with the growth and ethanol fermentation of . The current study was focused on the impact of cofactor availability derived intracellular redox perturbation on furfural tolerance. Here, three strategies were employed in cofactor conversion in : (1) heterologous expression of NADH dehydrogenase () from which catalyzed the NADH to NAD and increased the cellular sensitivity to furfural, (2) overexpression of , and genes responsible for the interconversion of NADPH and NADP, which enhanced the furfural tolerance, (3) expression of NAD(P) transhydrogenase () and NAD kinase () which showed a little impact on furfural tolerance. Besides, a substantial redistribution of metabolic fluxes was also observed with the expression of cofactor-related genes. These results indicated that NADPH-based intracellular redox perturbation plays a key role in furfural tolerance, which suggested single-gene manipulation as an effective strategy for enhancing tolerance and subsequently achieving higher ethanol titer using lignocellulosic hydrolysate.
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http://dx.doi.org/10.3389/fbioe.2020.00615DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7324476PMC
June 2020

Editorial for the special issue on "Metabolic engineering".

Biotechnol Appl Biochem 2020 Jan;67(1):5-6

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, People's Republic of China.

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http://dx.doi.org/10.1002/bab.1904DOI Listing
January 2020

Potential of as an electricity producer in ethanol production.

Biotechnol Biofuels 2020 5;13:36. Epub 2020 Mar 5.

1State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Sciences of Ministry of Education, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai, 200240 China.

Background: Microbial fuel cell (MFC) convokes microorganism to convert biomass into electricity. However, most well-known electrogenic strains cannot directly use glucose to produce valuable products. , a promising bacterium for ethanol production, owns special Entner-Doudoroff pathway with less ATP and biomass produced and the low-energy coupling respiration, making a potential exoelectrogen.

Results: A glucose-consuming MFC is constructed by inoculating . The electricity with power density 2.0 mW/m is derived from the difference of oxidation-reduction potential (ORP) between anode and cathode chambers. Besides, two-type electricity generation is observed as glucose-independent process and glucose-dependent process. For the sake of enhancing MFC efficiency, extracellular and intracellular strategies are implemented. Biofilm removal and addition of -type cytochrome benefit electricity performance and Tween 80 accelerates the electricity generation. Perturbation of cellular redox balance compromises the electricity output, indicating that redox homeostasis is the principal requirement to reach ideal voltage.

Conclusion: This study identifies potential feature of electricity activity for and provides multiple strategies to enhance the electricity output. Therefore, additional electricity generation will benefit the techno-economic viability of the commercial bulk production for biochemicals or biofuels in an efficient and environmentally sustainable manner.
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http://dx.doi.org/10.1186/s13068-020-01672-5DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7057670PMC
March 2020

Identification of a novel repressor encoded by the putative gene ctf1 for cellulase biosynthesis in Trichoderma reesei through artificial zinc finger engineering.

Biotechnol Bioeng 2020 06 15;117(6):1747-1760. Epub 2020 Mar 15.

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Science, and School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai, China.

Strains from Trichoderma reesei have been used for cellulase production with a long history. It has been well known that cellulase biosynthesis by the fungal species is controlled through regulators, and elucidation of their regulation network is of great importance for engineering T. reesei with robust cellulase production. However, progress in this regard is still very limited. In this study, T. reesei RUT-C30 was transformed with an artificial zinc finger protein (AZFP) library, and the mutant T. reesei M2 with improved cellulase production was screened. Compared to its parent strain, the filter paper activity and endo-β-glucanase activity in cellulases produced by T. reesei M2 increased 67.2% and 35.3%, respectively. Analysis by quantitative reverse transcription polymerase chain reaction indicated significant downregulation of the putative gene ctf1 in T. reesei M2, and its deletion mutants were thus developed for further studies. An increase of 36.9% in cellulase production was observed in the deletion mutants, but when ctf1 was constitutively overexpressed in T. reesei RUT-C30 under the control of the strong pdc1 promoter, cellulase production was substantially compromised. Comparative transcriptomic analysis revealed that the deletion of ctf1 upregulated transcription of gene encoding the regulator VIB1, but downregulated transcription of gene encoding another regulator RCE1, which consequently upregulated genes encoding the transcription factors XYR1 and ACE3 for the activation of genes encoding cellulolytic enzymes. As a result, ctf1 was characterized as a gene encoding a repressor for cellulase production in T. reesei RUT-C30, which is significant for further elucidating molecular mechanism underlying cellulase biosynthesis by the fungal species for rational design to develop robust strains for cellulase production. And in the meantime, AZFP transformation was validated to be an effective strategy for identifying functions of putative genes in the genome of T. reesei.
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http://dx.doi.org/10.1002/bit.27321DOI Listing
June 2020

Subcellular Performance of Nanoparticles in Cancer Therapy.

Int J Nanomedicine 2020 5;15:675-704. Epub 2020 Feb 5.

Institute of Biomaterials and Tissue Engineering, Huaqiao University, Xiamen, Fujian 361021, People's Republic of China.

With the advent of nanotechnology, various modes of traditional treatment strategies have been transformed extensively owing to the advantageous morphological, physiochemical, and functional attributes of nano-sized materials, which are of particular interest in diverse biomedical applications, such as diagnostics, sensing, imaging, and drug delivery. Despite their success in delivering therapeutic agents, several traditional nanocarriers often end up with deprived selectivity and undesired therapeutic outcome, which significantly limit their clinical applicability. Further advancements in terms of improved selectivity to exhibit desired therapeutic outcome toward ablating cancer cells have been predominantly made focusing on the precise entry of nanoparticles into tumor cells via targeting ligands, and subsequent delivery of therapeutic cargo in response to specific biological or external stimuli. However, there is enough room intracellularly, where diverse small-sized nanomaterials can accumulate and significantly exert potentially specific mechanisms of antitumor effects toward activation of precise cancer cell death pathways that can be explored. In this review, we aim to summarize the intracellular pathways of nanoparticles, highlighting the principles and state of their destructive effects in the subcellular structures as well as the current limitations of conventional therapeutic approaches. Next, we give an overview of subcellular performances and the fate of internalized nanoparticles under various organelle circumstances, particularly endosome or lysosome, mitochondria, nucleus, endoplasmic reticulum, and Golgi apparatus, by comprehensively emphasizing the unique mechanisms with a series of interesting reports. Moreover, intracellular transformation of the internalized nanoparticles, prominent outcome and potential affluence of these interdependent subcellular components in cancer therapy are emphasized. Finally, we conclude with perspectives with a focus on the contemporary challenges in their clinical applicability.
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http://dx.doi.org/10.2147/IJN.S226186DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7008395PMC
May 2020

Highly Porous Microcarriers for Minimally Invasive In Situ Skeletal Muscle Cell Delivery.

Small 2019 06 8;15(25):e1901397. Epub 2019 May 8.

Institute of Biomaterials and Tissue Engineering, Huaqiao University, Xiamen, 361021, P. R. China.

Microscale cell carriers have recently garnered enormous interest in repairing tissue defects by avoiding substantial open surgeries using implants for tissue regeneration. In this study, the highly open porous microspheres (HOPMs) are fabricated using a microfluidic technique for harboring proliferating skeletal myoblasts and evaluating their feasibility toward cell delivery application in situ. These biocompatible HOPMs with particle sizes of 280-370 µm possess open pores of 10-80 µm and interconnected paths. Such structure of the HOPMs conveniently provide a favorable microenvironment, where the cells are closely arranged in elongated shapes with the deposited extracellular matrix, facilitating cell adhesion and proliferation, as well as augmented myogenic differentiation. Furthermore, in vivo results in mice confirm improved cell retention and vascularization, as well as partial myoblast differentiation. These modular cell-laden microcarriers potentially allow for in situ tissue construction after minimally invasive delivery providing a convenient means for regeneration medicine.
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http://dx.doi.org/10.1002/smll.201901397DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6750270PMC
June 2019

3D-Printing of Microfibrous Porous Scaffolds Based on Hybrid Approaches for Bone Tissue Engineering.

Polymers (Basel) 2018 Jul 23;10(7). Epub 2018 Jul 23.

Institute of Biomaterials and Tissue Egineering, Huaqiao University, Xiamen 361021, China.

In recent times, tremendous progress has been evidenced by the advancements in various methods of generating three-dimensional (3D) porous scaffolds. However, the applicability of most of the traditional approaches intended for generating these biomimetic scaffolds is limited due to poor resolution and strict requirements in choosing materials. In this work, we fabricated 3D porous scaffolds based on the composite inks of gelatin (Gel), nano-hydroxyapatite (n-HA), and poly(lactide--glycolide) (PLGA) using an innovative hybrid strategy based on 3D printing and freeze-drying technologies for bone tissue engineering. Initially, the PLGA scaffolds were printed using the 3D printing method, and they were then coated with the Gel/n-HA complex, yielding the Gel/n-HA/PLGA scaffolds. These Gel/n-HA/PLGA scaffolds with exceptional biodegradation, mechanical properties, and biocompatibility have enabled osteoblasts (MC3T3-E1) for their convenient adhesion as a layer and have efficiently promoted their growth, as well as differentiation. We further demonstrated the bone growth by measuring the particular biomarkers that act as key players in the ossification process (i.e., alkaline phosphatase (ALP), osteocalcin (OC), and collagen type-I (COL-I)) and the total proteins of the MC3T3-E1 cells. We anticipate that the convenient generation of highly porous 3D scaffolds based on Gel/n-HA/PLGA fabricated through an innovative combinatorial approach of 3D printing technology and freeze-drying methods may undoubtedly find widespread applications in regenerative medicine.
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http://dx.doi.org/10.3390/polym10070807DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6404034PMC
July 2018

[Comparison of soil organic matter, bulk density and clay content in small watersheds under different ecological managements of Loess Plateau, China].

Ying Yong Sheng Tai Xue Bao 2019 Feb;30(2):370-378

State Key Laboratory of Soil Erosion and Dry-land Farming on the Loess Plateau, Institute of Soil and Water Conservation, Northwest A&F University, Yangling 712100, Shaanxi, China.

To explore the effects of small watersheds with different ecological managements on soil properties, the spatial differences of soil organic matter (SOM), bulk density (BD), and clay content (CC) in the four facets, including slope aspect, slope position, zone, and soil layer, were analyzed between Yangjiagou (YJG, artificial Robinia pseudoacacia forest watershed) and Dongzhuanggou (DZG, closed grassland watershed). The results showed that SOM, BD and CC were 12.78 g·kg, 1.24 g·cm, 19.2% for YJG and 11.13 g·kg, 1.21 g·cm, 18.2% for DZG, respectively. The values for YJG were slightly higher than those for DZG, but the difference was insignificant. All indices in the east slope were bigger than those in the west slope. Across different slope positions, the variation of BD was small, SOM and CC showed increasing trends from top to bottom. BD and CC declined downward the watershed, whereas SOM changed in an opposite trend. From the soil surface down to 60 cm soil depth, BD and CC increased and SOM decreased. The spatial sensitivity followed CC > SOM > BD, and the effects of the spatial factors can be ordered as soil layer > zone > slope aspect > slope position. There were significant differences in CC of the upper reaches, BD and CC of the middle reaches between the two basins. The sensitivity of each index to slope position, zone and soil layer in YJG was lower than that in DZG.
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http://dx.doi.org/10.13287/j.1001-9332.201902.013DOI Listing
February 2019

Engineering Zymomonas mobilis for Robust Cellulosic Ethanol Production.

Trends Biotechnol 2019 09 13;37(9):960-972. Epub 2019 Mar 13.

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, and School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China. Electronic address:

Great effort has been devoted to engineering Saccharomyces cerevisiae with pentose metabolism through the oxido-reductase pathway for cellulosic ethanol production, but intrinsic cofactor imbalance is observed, which substantially compromises ethanol yield. Zymomonas mobilis not only can be engineered for pentose metabolism through the isomerase pathway without cofactor imbalance but also metabolizes sugar through the Entner-Doudoroff pathway with less ATP and biomass produced for more sugar to be used for ethanol production. Moreover, the availabilities of genome sequence information for multiple Z. mobilis strains and advanced genetics tools have laid a solid foundation for engineering this species, and the self-flocculation of the bacterial cells also presents significant advantages for bioprocess engineering. Here, we highlight some of recent advances in these aspects.
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http://dx.doi.org/10.1016/j.tibtech.2019.02.002DOI Listing
September 2019

Cellulosic ethanol production: Progress, challenges and strategies for solutions.

Biotechnol Adv 2019 May - Jun;37(3):491-504. Epub 2019 Mar 5.

State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic & Developmental Science and School of Life Science and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China; School of Life Science and Biotechnology, Dalian University of Technology, Dalian 116024, China. Electronic address:

Lignocellulosic biomass is a sustainable feedstock for fuel ethanol production, but it is characterized by low mass and energy densities, and distributed production with relatively small scales is more suitable for cellulosic ethanol, which can better balance cost for the feedstock logistics. Lignocellulosic biomass is recalcitrant to degradation, and pretreatment is needed, but more efficient pretreatment technologies should be developed based on an in-depth understanding of its biosynthesis and regulation for engineering plant cell walls with less recalcitrance. Simultaneous saccharification and co-fermentation has been developed for cellulosic ethanol production, but the concept has been mistakenly defined, since the saccharification and co-fermentation are by no means simultaneous. Lignin is unreactive, which not only occupies reactor spaces during the enzymatic hydrolysis of the cellulose component and ethanol fermentation thereafter, but also requires extra mixing, making high solid loading difficult for lignocellulosic biomass and ethanol titers substantially compromised, which consequently increases energy consumption for ethanol distillation and stillage discharge, presenting another challenge for cellulosic ethanol production. Pentose sugars released from the hydrolysis of hemicelluloses are not fermentable with Saccharomyces cerevisiae used for ethanol production from sugar- and starch-based feedstocks, and engineering the brewing yeast and other ethanologenic species such as Zymomonas mobilis with pentose metabolism has been performed within the past decades. However strategies for the simultaneous co-fermentation of pentose and hexose sugars that have been pursued overwhelmingly for strain development might be modified for robust ethanol production. Finally, unit integration and system optimization are needed to maximize economic and environmental benefits for cellulosic ethanol production. In this article, we critically reviewed updated progress, and highlighted challenges and strategies for solutions.
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http://dx.doi.org/10.1016/j.biotechadv.2019.03.002DOI Listing
July 2019

N-Doped Sandwich-Structured [email protected]@Pt Interface with Ultralow Pt Loading for pH-Universal Hydrogen Evolution Reaction.

ACS Appl Mater Interfaces 2019 Jan 17;11(4):4047-4056. Epub 2019 Jan 17.

Designing a unique electrochemical interface to exhibit Pt-like activity and good stability is indispensable for the efficient hydrogen evolution reaction (HER). Herein, we synthesize well-defined [email protected]@Pt nanospheres with a sandwich-structured interface through a facile organic-inorganic pyrolysis and following reduction process. The obtained [email protected]@Pt heterostructures with ultralow Pt loading are composed of well-dispersed MoC nanoparticles (NPs) inner layer, N-doped carbon layer, and ultrafine Pt NPs outer layer. Electrochemical measurements demonstrate that [email protected]@Pt heterostructures not only exhibit superior HER activities than commercial Pt/C with small overpotentials of only 27, 47, and 25 mV to achieve a current density of 10 mA cm in acidic, alkaline, and neutral media, respectively, but also possess favorable long-term stability in pH-universal solution. The improved reaction kinetics of [email protected]@Pt heterostructures are mainly attributed to the unique sandwich-structured interface with well-defined MoC NPs encapsulated by carbon layers and Pt NPs well-dispersed on the carbon support, synergistic effects among MoC NPs, NC, and Pt NPs, high specific surface area, and N-doping into the catalysts. This facile approach not only provides a new pathway for preparing well-defined carbides but also gives insight into the development of low-Pt catalysts for the efficient HER.
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http://dx.doi.org/10.1021/acsami.8b20209DOI Listing
January 2019

Engineered pH-responsive hydrazone-carboxylate complexes-encapsulated 2D matrices for cathepsin-mediated apoptosis in cancer.

J Biomed Mater Res A 2019 06 28;107(6):1184-1194. Epub 2019 Feb 28.

College of Chemical Engineering, Huaqiao University, Xiamen, People's Republic of China.

Spurred by the current advancements in engineering various intelligent nanoparticle-based drug delivery systems, conjugation of drugs with the stimuli-responsive molecular switches has become one of the most efficient approaches to deliver a drug cargo in spatiotemporal controlled fashions. In this study, we fabricated an innovative pH-triggered hydrazone-carboxylate complex of doxorubicin (Dox), which was subsequently encapsulated in the layered double hydroxide (LDH) nanoparticles for effective cancer therapeutics. These two-dimensional (2D) biodegradable matrices efficiently delivered Dox by pH-triggered release in the acidic lysosomal environment and their subsequent escape to cytosol. Moreover, the delivered Dox molecules and high positively-charged surfaces of LDHs facilitated the cancer cell ablation via enhancing the cathepsins-mediated cell apoptosis assisted by free radical species generation. The critical advancements in the nanoparticle-based designs and substantial ablation of tumor cells through a free radical attack indicate that the designed pH-triggered drug composites can be used for efficient cancer therapeutics. © 2019 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 1184-1194, 2019.
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http://dx.doi.org/10.1002/jbm.a.36610DOI Listing
June 2019

Pt-C Interfaces Based on Electronegativity-Functionalized Hollow Carbon Spheres for Highly Efficient Hydrogen Evolution.

ACS Appl Mater Interfaces 2018 Dec 6;10(50):43561-43569. Epub 2018 Dec 6.

State Key Laboratory of Heavy Oil Processing, Institute of New Energy , China University of Petroleum (East China) , Qingdao 266580 , P. R. China.

The hydrogen evolution reaction activity of carbon-supported Pt catalyst is highly dependent on Pt-C interfaces. Herein, we focus on the relationships between Pt activity and N/O-functionalized hollow carbon sphere (HCS) substrate in acidic media. The electrochemical dissolution of Pt counter electrode is performed to prepare Pt nanoparticles in low loading. The N groups are beneficial for homogeneously sized Pt nanoparticles, whereas the O groups lead to aggregated nanoparticles. Moreover, the proper electronegativity of the N groups may enable capturing of protons to create proton-rich Pt-C interfaces and transfer them onto the Pt sites. The O groups may also capture protons by hydrogen bonding, but the subsequent release of protons is more difficult due to a stronger electronegativity and result in an inferior Pt activity. Consequently, the N-doped HCS with a low Pt loading (1.7 μg cm and 0.05 wt %) possesses a higher intrinsic activity compared with Pt on O-doped HCS. Moreover, it outperforms the commercial 20% Pt/C with a stable operation for 12 h. This work may provide suggestions for constructing the advantageous Pt-C interfaces by proper functional groups for high catalytic efficiencies.
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http://dx.doi.org/10.1021/acsami.8b10845DOI Listing
December 2018

Electrochemical Corrosion Engineering for Ni-Fe Oxides with Superior Activity toward Water Oxidation.

ACS Appl Mater Interfaces 2018 Dec 27;10(49):42217-42224. Epub 2018 Nov 27.

State Key Laboratory of Heavy Oil Processing, Institute of New Energy , China University of Petroleum (East China) , Qingdao 266580 , PR China.

The traditional synthesis for bimetallic-based electrocatalysts is challengeable for fine composition and elemental distribution because of the uncontrollable growth speed of nanostructures utilizing metal salt precursors. Herein, a unique electrochemical corrosion engineering strategy is developed via electrochemically transforming metal solid substrates (iron foil and nickel foam) into a highly active Ni-Fe oxide film for oxygen evolution, rather than directly utilizing metal ion precursors. This synthesis involves electrochemical corrosion of a Fe foil in an aqueous electrolyte along with electrochemical passivation of Ni foam (NF). The released trace Fe ions gradually incorporate into passivated NF surfaces to construct Ni-Fe oxide film and crucially improve composition distribution in the catalyst film. As a result, the resulted film with an ultralow mass loading (0.22 mg cm) delivers large current densities of 500 mA cm at overpotential of only 270 mV in 6.0 M KOH at 60 °C, outperforming many reported NiFe catalysts requiring much higher mass loadings. More interestingly, the as-prepared catalyst almost reaches the standard (500 mA cm within the overpotential of 300 mV) in commercial water electrolysis with long-term stability for at least 10 h. This work may provide a unique synthesis strategy for nonprecious transition-metal catalysts for desirable water splitting and can be expanded to many other electrocatalysis systems.
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http://dx.doi.org/10.1021/acsami.8b13267DOI Listing
December 2018

Characterization and Preliminary Biological Evaluation of 3D-Printed Porous Scaffolds for Engineering Bone Tissues.

Materials (Basel) 2018 Sep 26;11(10). Epub 2018 Sep 26.

Institute of Biomaterials and Tissue Engineering, Huaqiao University, Xiamen 361021, China.

Some basic requirements of bone tissue engineering include cells derived from bone tissues, three-dimensional (3D) scaffold materials, and osteogenic factors. In this framework, the critical architecture of the scaffolds plays a crucial role to support and assist the adhesion of the cells, and the subsequent tissue repairs. However, numerous traditional methods suffer from certain drawbacks, such as multi-step preparation, poor reproducibility, high complexity, difficulty in controlling the porous architectures, the shape of the scaffolds, and the existence of solvent residue, which limits their applicability. In this work, we fabricated innovative poly(lactic--glycolic acid) (PLGA) porous scaffolds, using 3D-printing technology, to overcome the shortcomings of traditional approaches. In addition, the printing parameters were critically optimized for obtaining scaffolds with normal morphology, appropriate porous architectures, and sufficient mechanical properties, for the accommodation of the bone cells. Various evaluation studies, including the exploration of mechanical properties (compressive strength and yield stress) for different thicknesses, and change of structure (printing angle) and porosity, were performed. Particularly, the degradation rate of the 3D scaffolds, printed in the optimized conditions, in the presence of hydrolytic, as well as enzymatic conditions were investigated. Their assessments were evaluated using the thermal gravimetric analyzer (TGA), differential scanning calorimetry (DSC), and gel permeation chromatography (GPC). These porous scaffolds, with their biocompatibility, biodegradation ability, and mechanical properties, have enabled the embryonic osteoblast precursor cells (MC3T3-E1), to adhere and proliferate in the porous architectures, with increasing time. The generation of highly porous 3D scaffolds, based on 3D printing technology, and their critical evaluation, through various investigations, may undoubtedly provide a reference for further investigations and guide critical optimization of scaffold fabrication, for tissue regeneration.
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http://dx.doi.org/10.3390/ma11101832DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6213437PMC
September 2018

Chemiluminescence imaging of UVA induced reactive oxygen species in mouse skin using L-012 as a probe.

Free Radic Res 2018 Dec 11;52(11-12):1424-1431. Epub 2018 Sep 11.

a School of Biomedical Engineering , Shanghai Jiao Tong University , Shanghai , China.

Reactive oxygen species (ROS) caused by ultraviolet A (UVA) can be reduced by treating with antioxidants and photoprotective reagents. Here we reported a real-time chemiluminescence (CL) imaging method which was simple, non-invasive and sensitive to evaluate UVA-induced ROS generation and the efficacy of sunscreens and antioxidants in vivo. The in vitro experiments indicated that l-ascorbic acid, live SPSC01 yeast, and its intracellular metabolites can suppress the intensity of CL signals in the presence of hydrogen peroxide, which proved the good antioxidant ability of them. Meanwhile, we used 8-amino-5-chloro-7-phenylpyrido[3,4-d] pyridazine-1,4(2H,3H) dione (L-012) as a high sensitive CL probe for in vivo imaging of ROS generated by UVA irradiation. The CL intensity was reduced after treated with l-ascorbic acid and SPSC01 yeast intracellular metabolites, consistent with the in vitro results. Additionally, the in vivo protective capability of two azobenzene compounds as sunscreens was confirmed further through the suppression of CL signals of UVA-induced ROS in mouse skin by this method.
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http://dx.doi.org/10.1080/10715762.2018.1500019DOI Listing
December 2018

Modular engineering to increase intracellular NAD(H/) promotes rate of extracellular electron transfer of Shewanella oneidensis.

Nat Commun 2018 09 7;9(1):3637. Epub 2018 Sep 7.

Key Laboratory of Systems Bioengineering (Ministry of Education), SynBio Research Platform, Collaborative Innovation Centre of Chemical Science and Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin, 300072, PR China.

The slow rate of extracellular electron transfer (EET) of electroactive microorganisms remains a primary bottleneck that restricts the practical applications of bioelectrochemical systems. Intracellular NAD(H/) (i.e., the total level of NADH and NAD) is a crucial source of the intracellular electron pool from which intracellular electrons are transferred to extracellular electron acceptors via EET pathways. However, how the total level of intracellular NAD(H/) impacts the EET rate in Shewanella oneidensis has not been established. Here, we use a modular synthetic biology strategy to redirect metabolic flux towards NAD biosynthesis via three modules: de novo, salvage, and universal biosynthesis modules in S. oneidensis MR-1. The results demonstrate that an increase in intracellular NAD(H/) results in the transfer of more electrons from the increased oxidation of the electron donor to the EET pathways of S. oneidensis, thereby enhancing intracellular electron flux and the EET rate.
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http://dx.doi.org/10.1038/s41467-018-05995-8DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6128845PMC
September 2018
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