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Article ; Online: Hyper-production of porcine contagious pleuropneumonia subunit vaccine proteins in Escherichia coli by developing a bicistronic T7 expression system.

Sun, Manman / Gao, Alex Xiong / Li, An / Ledesma-Amaro, Rodrigo / Wang, Peng / Chen, Wenchao / Bai, Zhonghu / Liu, Xiuxia

Biotechnology journal

2024 Volume 19, Issue 1, Page(s) e2300187

Abstract: The ApxII toxin and the outer membrane lipoprotein (Oml) of Actinobacillus pleuropneumoniae are important vaccine antigens against porcine contagious pleuropneumonia (PCP), a prevalent infectious disease affecting the swine industry worldwide. Previous ... ...

Abstract	The ApxII toxin and the outer membrane lipoprotein (Oml) of Actinobacillus pleuropneumoniae are important vaccine antigens against porcine contagious pleuropneumonia (PCP), a prevalent infectious disease affecting the swine industry worldwide. Previous studies have reported the recombinant expression of ApxII and Oml in Escherichia coli; however, their yields were not satisfactory. Here, we aimed to enhance the production of ApxII and Oml by constructing a bicistronic expression system based on the widely used T7 promoter. To create efficient T7 bicistronic expression cassettes, 16 different fore-cistron sequences were introduced downstream of the T7 promoter. The expression of three vaccine antigens Oml1, Oml7, and ApxII in the four strongest bicistronic vectors were enhanced compared to the monocistronic control. Further optimization of the fermentation conditions in micro-well plates (MWP) led to improved production. Finally, the production yields reached unprecedented levels of 2.43 g L
MeSH term(s)	Swine ; Animals ; Bacterial Proteins ; Escherichia coli/genetics ; Pleuropneumonia, Contagious/prevention & control ; Recombinant Proteins/genetics ; Actinobacillus Infections/prevention & control ; Vaccines, Subunit/genetics
Chemical Substances	Bacterial Proteins ; Recombinant Proteins ; Vaccines, Subunit
Language	English
Publishing date	2024-01-04
Publishing country	Germany
Document type	Journal Article
ZDB-ID	2221885-3
ISSN	1860-7314 ; 1860-6768
ISSN (online)	1860-7314
ISSN	1860-6768
DOI	10.1002/biot.202300187
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Article ; Online: High-throughput process development from gene cloning to protein production.

Sun, Manman / Gao, Alex Xiong / Liu, Xiuxia / Yang, Yankun / Ledesma-Amaro, Rodrigo / Bai, Zhonghu

Microbial cell factories

2023 Volume 22, Issue 1, Page(s) 182

Abstract: In the post-genomic era, the demand for faster and more efficient protein production has increased, both in public laboratories and industry. In addition, with the expansion of protein sequences in databases, the range of possible enzymes of interest for ...

Abstract	In the post-genomic era, the demand for faster and more efficient protein production has increased, both in public laboratories and industry. In addition, with the expansion of protein sequences in databases, the range of possible enzymes of interest for a given application is also increasing. Faced with peer competition, budgetary, and time constraints, companies and laboratories must find ways to develop a robust manufacturing process for recombinant protein production. In this review, we explore high-throughput technologies for recombinant protein expression and present a holistic high-throughput process development strategy that spans from genes to proteins. We discuss the challenges that come with this task, the limitations of previous studies, and future research directions.
MeSH term(s)	Cloning, Molecular ; Amino Acid Sequence ; Genomics ; Laboratories ; Recombinant Proteins/genetics
Chemical Substances	Recombinant Proteins
Language	English
Publishing date	2023-09-15
Publishing country	England
Document type	Journal Article ; Review
ZDB-ID	2091377-1
ISSN	1475-2859 ; 1475-2859
ISSN (online)	1475-2859
ISSN	1475-2859
DOI	10.1186/s12934-023-02184-1
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Article ; Online: Flavonoids from Seabuckthorn (

Xia, Chen-Xi / Gao, Alex Xiong / Zhu, Yue / Dong, Tina Ting-Xia / Tsim, Karl Wah-Keung

Food & function

2023 Volume 14, Issue 16, Page(s) 7426–7438

Abstract: Seabuckthorn ( ...

Abstract	Seabuckthorn (
MeSH term(s)	Humans ; Mice ; Animals ; Flavonoids/pharmacology ; Hippophae ; Depressive Disorder, Major/drug therapy ; Gastrointestinal Microbiome ; Depression/drug therapy
Chemical Substances	Flavonoids
Language	English
Publishing date	2023-08-14
Publishing country	England
Document type	Journal Article
ZDB-ID	2612033-1
ISSN	2042-650X ; 2042-6496
ISSN (online)	2042-650X
ISSN	2042-6496
DOI	10.1039/d3fo01332d
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Article ; Online: Flavonoids from Seabuckthorn (Hippophae rhamnoides L.) mimic neurotrophic functions in inducing neurite outgrowth in cultured neurons: Signaling via PI3K/Akt and ERK pathways.

Xia, Chen-Xi / Gao, Alex Xiong / Dong, Tina Ting-Xia / Tsim, Karl Wah-Keung

Phytomedicine : international journal of phytotherapy and phytopharmacology

2023 Volume 115, Page(s) 154832

Abstract: Background: Various brain disorders, including neurodegenerative diseases and major depressive disorders, threaten an increasing number of patients. Seabuckthorn, a fruit from Hippophae rhamnoides L., is an example of "medicine food homology". The fruit ...

Abstract	Background: Various brain disorders, including neurodegenerative diseases and major depressive disorders, threaten an increasing number of patients. Seabuckthorn, a fruit from Hippophae rhamnoides L., is an example of "medicine food homology". The fruit has enriched flavonoids that reported to have benefits in treating cognitive disorders. However, the studies on potential functions of Seabuckthorn and/or its flavonoid-enriched fraction in treating neurodegenerative disorders are limited. Purpose: This study aimed to determine the ability and mechanism of the flavonoid-enriched fraction of Seabuckthorn (named as SBF) in mimicking the neurotrophic functions in inducing neurite outgrowth of cultured neurons. Methods: Cultured PC12 cell line, SH-SY5Y cell line and primary neurons (cortical and hippocampal neurons isolated from E17-19 SD rat embryos) were the employed models to evaluate SBF in inducing neurite outgrowth by comparing to the effects of NGF and BDNF. Immuno-fluorescence staining was applied to identify the morphological change during the neuronal differentiation. Luciferase assay was utilized for analyzing the transcriptional regulation of neurofilaments and cAMP/CREB-mediated gene. Western blot assay was conducted to demonstrate the expressions of neurofilaments and phosphorylated proteins. Results: The application of SBF induced neuronal cell differentiation, and this differentiating activation was blocked by the inhibitors of PI3K/Akt and ERK pathways. Additionally, SBF showed synergy with neurotrophic factors in stimulating the neurite outgrowth of cultured neurons. Moreover, the major flavonoids within SBF, i.e., isorhamnetin, quercetin and kaempferol, could account for the neurotrophic activities of SBF. Conclusion: Seabuckthorn flavonoids mimicked neurotrophic functions in inducing neuronal cell differentiation via activating PI3K/Akt and ERK pathways. The results suggest the beneficial functions of Seabuckthorn as a potential health food supplement in treating various brain disorders, e.g., neurodegenerative diseases.
MeSH term(s)	Rats ; Humans ; Animals ; MAP Kinase Signaling System ; Proto-Oncogene Proteins c-akt/metabolism ; Phosphatidylinositol 3-Kinases/metabolism ; Hippophae ; Flavonoids/pharmacology ; Flavonoids/therapeutic use ; Neurites/metabolism ; Depressive Disorder, Major/metabolism ; Rats, Sprague-Dawley ; Neuroblastoma/metabolism ; Neurons ; Neuronal Outgrowth ; Neurodegenerative Diseases/drug therapy
Chemical Substances	Proto-Oncogene Proteins c-akt (EC 2.7.11.1) ; Phosphatidylinositol 3-Kinases (EC 2.7.1.-) ; Flavonoids
Language	English
Publishing date	2023-04-18
Publishing country	Germany
Document type	Journal Article
ZDB-ID	1205240-1
ISSN	1618-095X ; 0944-7113
ISSN (online)	1618-095X
ISSN	0944-7113
DOI	10.1016/j.phymed.2023.154832
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Article ; Online: The neurotrophic activities of brain-derived neurotrophic factor are potentiated by binding with apigenin, a common flavone in vegetables, in stimulating the receptor signaling.

Gao, Alex Xiong / Xia, Tracy Chen-Xi / Lin, Lish Sheng-Ying / Dong, Tina Ting-Xia / Tsim, Karl Wah-Keung

CNS neuroscience & therapeutics

2023 Volume 29, Issue 10, Page(s) 2787–2799

Abstract: Aims: We aimed to identify the neurotrophic activities of apigenin (4',5,7-trihydroxyflavone) via its coordination with brain-derived neurotrophic factor (BNDF) and an elevated signaling of tyrosine kinase receptor B (Trk B receptor).: Methods: The ... ...

Abstract	Aims: We aimed to identify the neurotrophic activities of apigenin (4',5,7-trihydroxyflavone) via its coordination with brain-derived neurotrophic factor (BNDF) and an elevated signaling of tyrosine kinase receptor B (Trk B receptor). Methods: The direct binding of apigenin to BDNF was validated by ultrafiltration and biacore assay. Neurogenesis, triggered by apigenin and/or BDNF, was determined in cultured SH-SY5Y cells and rat cortical neurons. The amyloid-beta (Aβ) Results: Apigenin and BDNF synergistically maintained the cell viability and promoted neurite outgrowth of cultured neurons. In addition, the BDNF-induced neurogenesis of cultured neurons was markedly potentiated by applied apigenin, including the induced expressions of neurofilaments, PSD-95 and synaptotagmin. Moreover, the synergy of apigenin and BDNF alleviated the (Aβ) Conclusion: Apigenin potentiates the neurotrophic activities of BDNF through direct binding, which may serve as a possible treatment for its curative efficiency in neurodegenerative diseases and depression.
MeSH term(s)	Rats ; Humans ; Animals ; Brain-Derived Neurotrophic Factor/metabolism ; Apigenin/pharmacology ; Vegetables/metabolism ; Neuroblastoma ; Receptor, trkB/metabolism ; Cells, Cultured ; Flavones/pharmacology
Chemical Substances	Brain-Derived Neurotrophic Factor ; Apigenin (7V515PI7F6) ; Receptor, trkB (EC 2.7.10.1) ; Flavones
Language	English
Publishing date	2023-04-26
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2423461-8
ISSN	1755-5949 ; 1755-5930
ISSN (online)	1755-5949
ISSN	1755-5930
DOI	10.1111/cns.14230
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Article ; Online: Leaderless Bicistronic Design for Precise and Reliable Control of Gene Expression in

Liu, Xiuxia / Sun, Manman / Gao, Alex Xiong / Ledesma-Amaro, Rodrigo / Fang, Qiuwu / Yang, Yankun / Bai, Zhonghu

ACS synthetic biology

2023 Volume 12, Issue 7, Page(s) 2157–2167

Abstract: In synthetic biology, the precise control of gene expression is challenging due to the limited orthogonality of expression elements. Here, to address this issue and improve the reusability of genetic elements, we developed a bicistronic expression ... ...

Abstract	In synthetic biology, the precise control of gene expression is challenging due to the limited orthogonality of expression elements. Here, to address this issue and improve the reusability of genetic elements, we developed a bicistronic expression cassette in
MeSH term(s)	Corynebacterium glutamicum/genetics ; Corynebacterium glutamicum/metabolism ; Promoter Regions, Genetic/genetics ; Gene Library ; Gene Expression ; Gene Expression Regulation, Bacterial/genetics
Language	English
Publishing date	2023-06-23
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2161-5063
ISSN (online)	2161-5063
DOI	10.1021/acssynbio.3c00246
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Article ; Online: The ethanolic extract of peanut shell attenuates the depressive-like behaviors of mice through modulation of inflammation and gut microbiota.

Gao, Alex Xiong / Xia, Tracy Chen-Xi / Peng, Zhi-Tian / Wu, Qi-Yun / Zhu, Yue / Dong, Tina Ting-Xia / Tsim, Karl Wah-Keung

Food research international (Ottawa, Ont.)

2023 Volume 168, Page(s) 112765

Abstract: Peanut shell is an agricultural byproduct being wasted on a large scale, which is in urgent need to be recycled. To fully utilize its pharmacological ingredients, e.g. luteolin, eriodyctiol, and 5,7-dihydroxychromone, we evaluated the curative effect of ... ...

Abstract	Peanut shell is an agricultural byproduct being wasted on a large scale, which is in urgent need to be recycled. To fully utilize its pharmacological ingredients, e.g. luteolin, eriodyctiol, and 5,7-dihydroxychromone, we evaluated the curative effect of ethanol extract deriving from peanut shell (PSE) in treating chronic unpredictable mild stress (CUMS)-induced depressive mice. The chronic stress lasted for 10 weeks, and PSE at 100-900 mg/kg/day was gavaged to mice in the last 2 weeks of modeling. The depressive behaviors were assessed by analyses of sucrose preference, tail suspension, and forced swimming. The brain injury was demonstrated by Hematoxylin and Eosin (H&E), Nissl body, and TdT-mediated dUTP nick end labeling (TUNEL) stainings in the mouse hippocampus. Biochemical indicators were analyzed, including levels of neurotrophic factors, neurotransmitters, stress hormones, and inflammatory mediators. The feces were collected for the 16S rDNA sequencing of gut microbiome. Administration of PSE improved the sucrose water consumption of depressive mice, while it decreased the immobile time in tail suspension and forced swimming tests. Meanwhile, the anti-depressive effect of PSE was supported by ameliorated histochemical staining, increased levels of neurotrophic factors and neurotransmitters, as well as down-regulated stress hormones. Furthermore, the treatment of PSE was able to mitigate the levels of inflammatory cytokines in brain, serum, and small intestine. Besides, the tight junction proteins, e.g., occludin and ZO-1, of gut showed elevated expressions, which coincided with the elevated abundance and diversity of gut microbiota upon PSE treatment. This study validated the therapeutic efficacy of PSE in fighting against depression, as well as its modulatory action on inflammation and gut microbiota, which promoted the recycling of this agricultural waste to be health supplements of added value.
MeSH term(s)	Mice ; Animals ; Depression/drug therapy ; Arachis ; Gastrointestinal Microbiome ; Inflammation ; Plant Extracts/pharmacology ; Nerve Growth Factors/pharmacology ; Hormones/pharmacology ; Ethanol ; Sucrose/pharmacology
Chemical Substances	Plant Extracts ; Nerve Growth Factors ; Hormones ; Ethanol (3K9958V90M) ; Sucrose (57-50-1)
Language	English
Publishing date	2023-03-27
Publishing country	Canada
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	1111695-x
ISSN	1873-7145 ; 0963-9969
ISSN (online)	1873-7145
ISSN	0963-9969
DOI	10.1016/j.foodres.2023.112765
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Article: The extract of peanut shell enhances neurite outgrowth of neuronal cells: Recycling of agricultural waste for development of nutraceutical products

Gao, Alex Xiong / Xiao, Jian / Xia, Tracy Chen-Xi / Dong, Tina Ting-Xia / Tsim, Karl Wah-Keung

Journal of functional foods. 2022 Apr., v. 91

2022

Abstract: Peanut shell is an agricultural waste having an urgent need to be utilized. The ethanolic extract of peanut shell extract, named as PSE, having enrichment of flavonoids with neurotrophic function, can develop as dietary supplements. In cultured PC12 ... ...

Abstract	Peanut shell is an agricultural waste having an urgent need to be utilized. The ethanolic extract of peanut shell extract, named as PSE, having enrichment of flavonoids with neurotrophic function, can develop as dietary supplements. In cultured PC12 cells, SH-SY5Y cells and rat primary neurons, the application of PSE increased the proportion of differentiated cells. In parallel, the neuronal differentiation markers, i.e. neurofilaments, were increased at both mRNA and protein levels under PSE treatment. In revealing the signaling cascade, extracellular signal-regulated kinase1/2 (ERK1/2) and protein kinase B (Akt) were activated by PSE. The PSE-induced cell differentiation was blocked by the kinase inhibitors, U0126 and LY294002. The phytochemicals within PSE, e.g. luteolin, eriodictyol and 5,7-dihydroxychromone, induced transcription of neurofilaments in cultures, which in combination could partially account for the neurotrophic functions of PSE. This result supports the waste product from peanut production could be recycled in developing high value nutraceutical products.
Keywords	agricultural wastes ; cell differentiation ; dietary supplements ; luteolin ; neurites ; non-specific serine/threonine protein kinase ; peanut hulls ; peanuts
Language	English
Dates of publication	2022-04
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	2511964-3
ISSN	1756-4646
ISSN	1756-4646
DOI	10.1016/j.jff.2022.105023
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Article: A Herbal Mixture of Sesami Semen Nigrum and Longan Arillus Induces Neurite Outgrowth in Cultured Neurons and Shows Anti-Depression in Chronic Mild Stress-Induced Rats.

Gao, Alex Xiong / Xia, Tracy Chen-Xi / Mak, Marvin Shing-Hung / Zhu, Kevin Yue / Dong, Tina Ting-Xia / Tsim, Karl Wah-Keung

Evidence-based complementary and alternative medicine : eCAM

2022 Volume 2022, Page(s) 8809546

Abstract: Medicinal food homology is referring to a group of food itself being considered as herbal medicine without a boundary of usage. Under the guidance of this food/medicine principle, the current study aims to develop anti-depressant from this food/medicine ... ...

Abstract	Medicinal food homology is referring to a group of food itself being considered as herbal medicine without a boundary of usage. Under the guidance of this food/medicine principle, the current study aims to develop anti-depressant from this food/medicine catalog. The herbal mixture of Sesami Semen Nigrum and Longan Arillus was evaluated in cultured PC12 rat pheochromocytoma cells, rat primary cortical neurons, and in chronic mild stress (CMS)-induced depressive rat model. The combination of two ethanolic extracts of Sesami Semen Nigrum and Longan Arillus in 1 : 1 ratio mimicked the function of nerve growth factor (NGF) and synergistically induced neurite outgrowth of PC12 cells. Besides, the expression and phosphorylation of tropomyosin receptor kinase A (TrkA) of the cultured cells were also elevated. This neurotrophic activity of herbal mixture was further supported by the increased expressions of biomarkers for neurogenesis and synaptogenesis in cortical neurons. Moreover, the depressed rats were soothed by the intake of herbal mixture, showing improved performance in behavior tests, as well as reversed levels of neurotransmitters and neurotrophic factors. Our results provide a new way to make full use of the current food/medicine resources, as to accelerate the development of therapeutics for depression.
Language	English
Publishing date	2022-06-16
Publishing country	United States
Document type	Journal Article
ZDB-ID	2171158-6
ISSN	1741-4288 ; 1741-427X
ISSN (online)	1741-4288
ISSN	1741-427X
DOI	10.1155/2022/8809546
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Article: Hypersecretion of OmlA antigen in Corynebacterium glutamicum through high-throughput based development process

Sun, Manman / Gao, Alex Xiong / Ledesma-Amaro, Rodrigo / Li, An / Wang, Rongbin / Nie, Jianqi / Zheng, Pei / Yang, Yankun / Bai, Zhonghu / Liu, Xiuxia

Applied microbiology and biotechnology. 2022 Apr., v. 106, no. 8

2022

Abstract: Outer membrane lipoprotein A (OmlA) is a vaccine antigen against porcine contagious pleuropneumonia (PCP), a disease severely affecting the swine industry. Here, we aimed to systematically potentiate the secretory production of OmlA in Corynebacterium ... ...

Abstract	Outer membrane lipoprotein A (OmlA) is a vaccine antigen against porcine contagious pleuropneumonia (PCP), a disease severely affecting the swine industry. Here, we aimed to systematically potentiate the secretory production of OmlA in Corynebacterium glutamicum (C. glutamicum), a widely used microorganism in the food industry, by establishing a holistic development process based on our high-throughput culture platform. The expression patterns, expression element combinations, medium composition, and induction conditions were comprehensively screened or optimized in microwell plates (MWPs), followed by fermentation parameter optimization in a 4 × 1 L parallel fermentation system (CUBER4). An unprecedented yield of 1.01 g/L OmlA was ultimately achieved in a 5-L bioreactor following the scaling-up strategy of fixed oxygen mass transfer coefficient (kLa), and the produced OmlA antigen showed well-protective immunity against Actinobacillus pleuropneumoniae challenge. This result provides a rapid and reliable pipeline to achieve the hyper-production of OmlA, and possibly other recombinant vaccines, in C. glutamicum. KEY POINTS: • Established a holistic development process and applied it to potentiate the secretion of OmlA. • The secretion of OmlA reached an unprecedented yield of 1.01 g/L. • The recombinant OmlA antigen induced efficient protective immunity.
Keywords	Actinobacillus pleuropneumoniae ; Corynebacterium glutamicum ; antigens ; bioreactors ; fermentation ; food industry ; hypersecretion ; immunity ; lipoproteins ; mass transfer ; microbiology ; oxygen ; pleuropneumonia ; pork industry ; swine
Language	English
Dates of publication	2022-04
Size	p. 2953-2967.
Publishing place	Springer Berlin Heidelberg
Document type	Article
ZDB-ID	392453-1
ISSN	1432-0614 ; 0171-1741 ; 0175-7598
ISSN (online)	1432-0614
ISSN	0171-1741 ; 0175-7598
DOI	10.1007/s00253-022-11918-x
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