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Article ; Online: In situ delivery of biobutyrate by probiotic Escherichia coli for cancer therapy

Chung-Jen Chiang / Yan-Hong Hong

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 17

Abstract: Abstract Butyrate has a bioactive function to reduce carcinogenesis. To achieve targeted cancer therapy, this study developed bacterial cancer therapy (BCT) with butyrate as a payload. By metabolic engineering, Escherichia coli Nissle 1917 (EcN) was ... ...

Abstract	Abstract Butyrate has a bioactive function to reduce carcinogenesis. To achieve targeted cancer therapy, this study developed bacterial cancer therapy (BCT) with butyrate as a payload. By metabolic engineering, Escherichia coli Nissle 1917 (EcN) was reprogrammed to synthesize butyrate (referred to as biobutyrate) and designated EcN-BUT. The adopted strategy includes construction of a synthetic pathway for biobutyrate and the rational design of central metabolism to increase the production of biobutyrate at the expense of acetate. With glucose, EcN-BUT produced primarily biobutyrate under the hypoxic condition. Furthermore, human colorectal cancer cell was administrated with the produced biobutyrate. It caused the cell cycle arrest at the G1 phase and induced the mitochondrial apoptosis pathway independent of p53. In the tumor-bearing mice, the injected EcN-BUT exhibited tumor-specific colonization and significantly reduced the tumor volume by 70%. Overall, this study opens a new avenue for BCT based on biobutyrate.
Keywords	Medicine ; R ; Science ; Q
Subject code	610
Language	English
Publishing date	2021-09-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Metabolic engineering of probiotic Escherichia coli for cytolytic therapy of tumors

Chung-Jen Chiang / Po-Han Huang

Scientific Reports, Vol 11, Iss 1, Pp 1-

2021 Volume 18

Abstract: Abstract Bacterial cancer therapy was developed using probiotic Escherichia coli Nissle 1917 (EcN) for medical intervention of colorectal cancer. EcN was armed with HlyE, a small cytotoxic protein, under the control of the araBAD promoter (PBAD). The ... ...

Abstract	Abstract Bacterial cancer therapy was developed using probiotic Escherichia coli Nissle 1917 (EcN) for medical intervention of colorectal cancer. EcN was armed with HlyE, a small cytotoxic protein, under the control of the araBAD promoter (PBAD). The intrinsic limitation of PBAD for the gene expression is known to be negated by glucose and afflicted with all-or-nothing induction in host bacteria. This issue was addressed by metabolic engineering of EcN to uncouple the glucose-mediated control circuit and the L-arabinose transport-induction loop and to block L-arabinose catabolism. As a result, the reprogrammed strain (designated EcNe) enabled efficient expression of HlyE in a temporal control manner. The HlyE production was insensitive to glucose and reached a saturated level in response to L-arabinose at 30–50 μM. Moreover, the administrated EcNe exhibited tumor-specific colonization with the tumor-to-organ ratio of 106:1. Equipped with HlyE, EcNe significantly caused tumor regression in mice xenografted with human colorectal cancer cells. Overall, this study proposes a new strategy for the bacteria-mediated delivery of therapeutic proteins to tumors.
Keywords	Medicine ; R ; Science ; Q
Subject code	570
Language	English
Publishing date	2021-03-01T00:00:00Z
Publisher	Nature Portfolio
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: The Herbal Cocktail GSYJ Attenuated Airway Inflammatory Cell Infiltration in a Chronic Asthmatic Mouse Model

Chung-Jen Chiang / Shu-Lun Chang / Li-Jen Lin

Evidence-Based Complementary and Alternative Medicine, Vol

2021 Volume 2021

Abstract: This study explored the potential therapeutic efficacy of GSYJ in attenuating asthma symptom severity and aimed to determine the immunomodulatory mechanism of GSYJ. A mouse model of chronic asthma induced by repeated Dermatophagoides pteronyssinus (Der p) ...

Abstract	This study explored the potential therapeutic efficacy of GSYJ in attenuating asthma symptom severity and aimed to determine the immunomodulatory mechanism of GSYJ. A mouse model of chronic asthma induced by repeated Dermatophagoides pteronyssinus (Der p) challenge was established. In addition, 30 minutes before Der p challenge, the mice were orally administered GSYJ (1 g/kg). The mice were sacrificed to evaluate inflammatory cell infiltration, collagen deposition in the lung, total IgE in serum, and expression profiles of various cytokines in bronchoalveolar lavage fluid (BALF) and various genes in lung tissue. Furthermore, 30 minutes after the addition of GSYJ to RAW264.7 cell cultures, 100 ng/ml LPS was added to evaluate the effect of the drug on the LPS-induced expression of genes, proteins, and transcription factors. GSYJ may regulate transcription factors (cJUN/IRF3/NF-κB) to decrease the expression of IL-1β, IL-6, RANTES, and iNOS in macrophages and affect the IL-12, IFN-γ, IL-5, and IL-6 levels in the BALF of mice to relieve asthma symptoms, such as inflammatory cell infiltration, hyperresponsiveness, and increased serum total IgE levels. Therefore, GSYJ has the potential to be developed into a drug treatment for chronic asthma.
Keywords	Other systems of medicine ; RZ201-999
Subject code	610
Language	English
Publishing date	2021-01-01T00:00:00Z
Publisher	Hindawi Limited
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Biocatalytic Conversion of Short-Chain Fatty Acids to Corresponding Alcohols in Escherichia coli

Li-Jen Lin / Mukesh Saini / Chung-Jen Chiang / Yun-Peng Chao

Processes, Vol 9, Iss 973, p

2021 Volume 973

Abstract: Advanced biofuels possess superior characteristics to serve for gasoline substitutes. In this study, a whole cell biocatalysis system was employed for production of short-chain alcohols from corresponding fatty acids. To do so, Escherichia coli strain ... ...

Abstract	Advanced biofuels possess superior characteristics to serve for gasoline substitutes. In this study, a whole cell biocatalysis system was employed for production of short-chain alcohols from corresponding fatty acids. To do so, Escherichia coli strain was equipped with a biocatalytic pathway consisting of endogenous atoDA and Clostridium acetobutylicum adhE2 . The strain was further reprogrammed to improve its biocatalytic activity by direction the glycolytic flux to acetyl-CoA and recycling acetate. The production of 1-propanol and n-pentanol were exemplified with the engineered strain. By substrate (glucose and propionate) feeding, the strain enabled production of 5.4 g/L 1-propanol with productivity reaching 0.15 g/L/h. In addition, the strain with a heavy inoculum was implemented for the n-pentanol production from n-pentanoic acid. The production titer and productivity finally attained 4.3 g/L and 0.86 g/L/h, respectively. Overall, the result indicates that this developed system is useful and effective for biocatalytic production of short-chain alcohols.
Keywords	short-chain fatty alcohols ; short-chain fatty acids ; biocatalysis ; Chemical technology ; TP1-1185 ; Chemistry ; QD1-999
Language	English
Publishing date	2021-05-01T00:00:00Z
Publisher	MDPI AG
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article: Metabolic engineering of Escherichia coli for production of n-butanol from crude glycerol

Saini, Mukesh / Ze Win Wang / Chung-Jen Chiang / Yun-Peng Chao

Biotechnology for biofuels. 2017 Dec., v. 10, no. 1

2017

Abstract: BACKGROUND: Crude glycerol in the waste stream of the biodiesel production process is an abundant and renewable resource. However, the glycerol-based industry is usually afflicted by the cost for refinement of crude glycerol. This issue can be addressed ... ...

Abstract	BACKGROUND: Crude glycerol in the waste stream of the biodiesel production process is an abundant and renewable resource. However, the glycerol-based industry is usually afflicted by the cost for refinement of crude glycerol. This issue can be addressed by developing a microbial process to convert crude glycerol to value-added chemicals. In this study, Escherichia coli was implemented for the production of n-butanol based on the reduced nature of glycerol. RESULTS: The central metabolism of E. coli was rewired to improve the efficiency of glycerol metabolism and provide the reductive need for n-butanol in E. coli. This was carried out in several steps by (1) forcing the glycolytic flux through the oxidation pathway of pyruvate, (2) directing the gluconeogenic flux into the oxidative pentose phosphate pathway, (3) enhancing the anaerobic catabolism for glycerol, and (4) moderately suppressing the tricarboxylic acid cycle. Under the microaerobic condition, the engineered strain enabled the production of 6.9 g/L n-butanol from 20 g/L crude glycerol. The conversion yield and the productivity reach 87% of the theoretical yield and 0.18 g/L/h, respectively. CONCLUSIONS: The approach by rational rewiring of metabolic pathways enables E. coli to synthesize n-butanol from glycerol in an efficient way. Our proposed strategies illustrate the feasibility of manipulating key metabolic nodes at the junction of the central catabolism. As a result, it renders the intracellular redox state adjustable for various purposes. Overall, the developed technology platform may be useful for the economic viability of the glycerol-related industry.
Keywords	Escherichia coli ; biodiesel ; butanol ; economic sustainability ; fuel production ; glycerol ; glycolysis ; metabolic engineering ; oxidation ; pentose phosphate cycle ; pyruvic acid ; tricarboxylic acid cycle ; value added
Language	English
Dates of publication	2017-12
Size	p. 173.
Publishing place	BioMed Central
Document type	Article
ZDB-ID	2421351-2
ISSN	1754-6834
ISSN	1754-6834
DOI	10.1186/s13068-017-0857-2
Database	NAL-Catalogue (AGRICOLA)

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Article: Systematic engineering of the central metabolism in Escherichia coli for effective production of n-butanol

Saini, Mukesh / Si-Yu Li / Ze Win Wang / Chung-Jen Chiang / Yun-Peng Chao

Biotechnology for biofuels. 2016 Dec., v. 9, no. 1

2016

Abstract: BACKGROUND: Microbes have been extensively explored for production of environment-friendly fuels and chemicals. The microbial fermentation pathways leading to these commodities usually involve many redox reactions. This makes the fermentative production ... ...

Abstract	BACKGROUND: Microbes have been extensively explored for production of environment-friendly fuels and chemicals. The microbial fermentation pathways leading to these commodities usually involve many redox reactions. This makes the fermentative production of highly reduced products challenging, because there is a limited NADH output from glucose catabolism. Microbial production of n-butanol apparently represents one typical example. RESULTS: In this study, we addressed the issue by adjustment of the intracellular redox state in Escherichia coli. This was initiated with strain BuT-8 which carries the clostridial CoA-dependent synthetic pathway. Three metabolite nodes in the central metabolism of the strain were targeted for engineering. First, the pyruvate node was manipulated by enhancement of pyruvate decarboxylation in the oxidative pathway. Subsequently, the pentose phosphate (PP) pathway was amplified at the glucose-6-phosphate (G6P) node. The pathway for G6P isomerization was further blocked to force the glycolytic flux through the PP pathway. It resulted in a growth defect, and the cell growth was later recovered by limiting the tricarboxylic acid cycle at the acetyl-CoA node. Finally, the resulting strain exhibited a high NADH level and enabled production of 6.1Â g/L n-butanol with a yield of 0.31Â g/g-glucose and a productivity of 0.21Â g/L/h. CONCLUSIONS: The production efficiency of fermentative products in microbes strongly depends on the intracellular redox state. This work illustrates the flexibility of pyruvate, G6P, and acetyl-CoA nodes at the junction of the central metabolism for engineering. In principle, high production of reduced products of interest can be achieved by individual or coordinated modulation of these metabolite nodes.
Keywords	Clostridium ; Escherichia coli ; NAD (coenzyme) ; acetyl coenzyme A ; butanol ; cell growth ; decarboxylation ; engineering ; fermentation ; fuels ; glucose ; glucose 6-phosphate ; glycolysis ; isomerization ; metabolites ; microorganisms ; pentoses ; phosphates ; products and commodities ; pyruvic acid ; redox reactions ; tricarboxylic acid cycle
Language	English
Dates of publication	2016-12
Size	p. 69.
Publishing place	BioMed Central
Document type	Article
ZDB-ID	2421351-2
ISSN	1754-6834
ISSN	1754-6834
DOI	10.1186/s13068-016-0467-4
Database	NAL-Catalogue (AGRICOLA)

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Article: Potential production platform of n-butanol in Escherichia coli

Saini, Mukesh / Chung-Jen Chiang / Min Hong Chen / Yun-Peng Chao

International Metabolic Engineering Society Metabolic engineering. 2015 Jan., v. 27

2015

Abstract: We proposed a potential production platform of n-butanol in Escherichia coli. First, a butyrate-conversion strain was developed by removal of undesired genes and recruiting endogenous atoDA and Clostridium adhE2. Consequently, this E. coli strain grown ... ...

Abstract	We proposed a potential production platform of n-butanol in Escherichia coli. First, a butyrate-conversion strain was developed by removal of undesired genes and recruiting endogenous atoDA and Clostridium adhE2. Consequently, this E. coli strain grown on the M9 mineral salt with yeast extract (M9Y) was shown to produce 6.2g/L n-butanol from supplemented butyrate at 36h. The molar conversion yield of n-butanol on butyrate reaches 92%. Moreover, the production platform was advanced by additional inclusion of a butyrate-producing strain. This strain was equipped with a pathway comprising atoDA and heterologous genes for the synthesis of butyrate. Without butyrate, the butyrate-conversion and the butyrate-producing strains were co-cultured in M9Y medium and produced 5.5g/L n-butanol from glucose at 24h. The production yield on glucose accounts for 69% of the theoretical yield. Overall, it indicates a promise of the developed platform for n-butanol production in E. coli.
Keywords	butanol ; butyrates ; Clostridium ; coculture ; Escherichia coli ; genes ; glucose ; yeast extract
Language	English
Dates of publication	2015-01
Size	p. 76-82.
Publishing place	Elsevier Inc.
Document type	Article
ZDB-ID	1470383-x
ISSN	1096-7184 ; 1096-7176
ISSN (online)	1096-7184
ISSN	1096-7176
DOI	10.1016/j.ymben.2014.11.001
Database	NAL-Catalogue (AGRICOLA)

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Article: Systematic Engineering of Escherichia coli for d-Lactate Production from Crude Glycerol

Wang, Zei Wen / Mukesh Saini / Li-Jen Lin / Chung-Jen Chiang / Yun-Peng Chao

Journal of agricultural and food chemistry. 2015 Nov. 4, v. 63, no. 43

2015

Abstract: Crude glycerol resulting from biodiesel production is an abundant and renewable resource. However, the impurities in crude glycerol usually make microbial fermentation problematic. This issue was addressed by systematic engineering of Escherichia coli ... ...

Abstract	Crude glycerol resulting from biodiesel production is an abundant and renewable resource. However, the impurities in crude glycerol usually make microbial fermentation problematic. This issue was addressed by systematic engineering of Escherichia coli for the production of d-lactate from crude glycerol. First, mgsA and the synthetic pathways of undesired products were eliminated in E. coli, rendering the strain capable of homofermentative production of optically pure d-lactate. To direct carbon flux toward d-lactate, the resulting strain was endowed with an enhanced expression of glpDâglpK in the glycerol catabolism and of a heterologous gene encoding d-lactate dehydrogenase. Moreover, the strain was evolved to improve its utilization of cruder glycerol and subsequently equipped with the FocA channel to export intracellular d-lactate. Finally, the fed-batch fermentation with two-phase culturing was carried out with a bioreactor. As a result, the engineered strain enabled production of 105 g/L d-lactate (99.9% optical purity) from 121 g/L crude glycerol at 40 h. The result indicates the feasibility of our approach to engineering E. coli for the crude glycerol-based fermentation.
Keywords	Escherichia coli ; batch fermentation ; biodiesel ; bioreactors ; carbon ; fuel production ; genes ; genetically engineered microorganisms ; glycerol ; lactic acid ; rendering
Language	English
Dates of publication	2015-1104
Size	p. 9583-9589.
Publishing place	American Chemical Society, Books and Journals Division
Document type	Article
ZDB-ID	241619-0
ISSN	1520-5118 ; 0021-8561
ISSN (online)	1520-5118
ISSN	0021-8561
DOI	10.1021/acs.jafc.5b04162
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Effects of spent mushroom compost meal on growth performance and meat characteristics of grower geese

Shen-Chang Chang / Min-Jung Lin / Yun-Peng Chao / Chung-Jen Chiang / Yu-Shine Jea / Tzu-Tai Lee

Revista Brasileira de Zootecnia, Vol 45, Iss 6, Pp 281-

2016 Volume 287

Abstract: ABSTRACT The objective of this study was to determine the effect of spent mushroom compost (SMC) meal on the growth performance and meat characteristics of geese. The SMC extracts contained 2.49±0.62 mg gallic acid equivalent/g dry weight (DW) and 1.08±0. ...

Abstract	ABSTRACT The objective of this study was to determine the effect of spent mushroom compost (SMC) meal on the growth performance and meat characteristics of geese. The SMC extracts contained 2.49±0.62 mg gallic acid equivalent/g dry weight (DW) and 1.08±0.15 mg quercetin equivalent/g DW of the total phenolic and flavonoid contents. A total of 120 White Roman geese, aged five weeks, were randomly distributed among 12 pens and fed a grower diet ad libitum during the growing period, with each pen containing five males and five females in a completely randomized design. Each treatment comprised three pens (total of 30 geese), including control (corn-soybean meal); supplementation with 5% SMC meal (5% SMC); supplementation with 10% SMC (10% SMC); and supplementation with 15% SMC (15% SMC), for eight weeks. The results revealed that the body weight of the 15% SMC group was significantly lower than that of the control group at the age of 12 weeks. There were no significant effects among the groups on feed conversion ratio or intake. The malondialdehyde content of the serum in the 15% SMC group was lower than that of the control group at week 12. There were no significant effects among the groups for blood biochemical parameters in grower geese at week 12. The color values of meats in the SMC groups were higher than those obtained with control group. The flavor and acceptability score of meats in the 5% SMC group were significantly higher than for the 15% SMC and control group. Supplementation with SMC at 5% in the diet has no adverse effects on the growth performance of grower geese. However, SMC meal at 5% in the diet favorably affects sensory attributes (meat flavor and acceptability).
Keywords	antioxidant ; meat sensory evaluation ; white Roman goose ; Animal culture ; SF1-1100 ; Agriculture ; S
Subject code	630
Language	English
Publishing date	2016-06-01T00:00:00Z
Publisher	Sociedade Brasileira de Zootecnia
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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Article ; Online: Effects of spent mushroom compost meal on growth performance and meat characteristics of grower geese

Shen-Chang Chang / Min-Jung Lin / Yun-Peng Chao / Chung-Jen Chiang / Yu-Shine Jea / Tzu-Tai Lee

Revista Brasileira de Zootecnia, Vol 45, Iss 6, Pp 336-

2016 Volume 344

Abstract	ABSTRACT The objective of this study was to determine the effect of spent mushroom compost (SMC) meal on the growth performance and meat characteristics of geese. The SMC extracts contained 2.49±0.62 mg gallic acid equivalent/g dry weight (DW) and 1.08±0.15 mg quercetin equivalent/g DW of the total phenolic and flavonoid contents. A total of 120 White Roman geese, aged five weeks, were randomly distributed among 12 pens and fed a grower diet ad libitum during the growing period, with each pen containing five males and five females in a completely randomized design. Each treatment comprised three pens (total of 30 geese), including control (corn-soybean meal); supplementation with 5% SMC meal (5% SMC); supplementation with 10% SMC (10% SMC); and supplementation with 15% SMC (15% SMC), for eight weeks. The results revealed that the body weight of the 15% SMC group was significantly lower than that of the control group at the age of 12 weeks. There were no significant effects among the groups on feed conversion ratio or intake. The malondialdehyde content of the serum in the 15% SMC group was lower than that of the control group at week 12. There were no significant effects among the groups for blood biochemical parameters in grower geese at week 12. The color values of meats in the SMC groups were higher than those obtained with control group. The flavor and acceptability score of meats in the 5% SMC group were significantly higher than for the 15% SMC and control group. Supplementation with SMC at 5% in the diet has no adverse effects on the growth performance of grower geese. However, SMC meal at 5% in the diet favorably affects sensory attributes (meat flavor and acceptability).
Keywords	antioxidant ; meat sensory evaluation ; white Roman goose ; Animal culture ; SF1-1100 ; Agriculture ; S
Subject code	630
Language	English
Publishing date	2016-06-01T00:00:00Z
Publisher	Sociedade Brasileira de Zootecnia
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

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