Metabolic engineering of Klebsiella pneumoniae for the de novo production of 2-butanol as a potential biofuel

2015 ◽  
Vol 197 ◽  
pp. 260-265 ◽  
Author(s):  
Zhen Chen ◽  
Yao Wu ◽  
Jinhai Huang ◽  
Dehua Liu
Keyword(s):  
Metabolic Engineering ◽  
Klebsiella Pneumoniae ◽  
De Novo

scholarly journals Metabolic engineering of Escherichia coli for de novo production of 3-phenylpropanol via retrobiosynthesis approach

2021 ◽  
Vol 20 (1) ◽  
Author(s):  
Zhenning Liu ◽  
Xue Zhang ◽  
Dengwei Lei ◽  
Bin Qiao ◽  
Guang-Rong Zhao
Keyword(s):  
Escherichia Coli ◽  
Metabolic Engineering ◽  
De Novo ◽  
Microbial Cell ◽  
Cell Factory ◽  
Phosphopantetheinyl Transferase ◽  
Chromosome Engineering ◽  
Microbial Cell Factory ◽  
E Coli ◽  
Artificial Pathway

Abstract Background 3-Phenylpropanol with a pleasant odor is widely used in foods, beverages and cosmetics as a fragrance ingredient. It also acts as the precursor and reactant in pharmaceutical and chemical industries. Currently, petroleum-based manufacturing processes of 3-phenypropanol is environmentally unfriendly and unsustainable. In this study, we aim to engineer Escherichia coli as microbial cell factory for de novo production of 3-phenypropanol via retrobiosynthesis approach. Results Aided by in silico retrobiosynthesis analysis, we designed a novel 3-phenylpropanol biosynthetic pathway extending from l-phenylalanine and comprising the phenylalanine ammonia lyase (PAL), enoate reductase (ER), aryl carboxylic acid reductase (CAR) and phosphopantetheinyl transferase (PPTase). We screened the enzymes from plants and microorganisms and reconstructed the artificial pathway for conversion of 3-phenylpropanol from l-phenylalanine. Then we conducted chromosome engineering to increase the supply of precursor l-phenylalanine and combined the upstream l-phenylalanine pathway and downstream 3-phenylpropanol pathway. Finally, we regulated the metabolic pathway strength and optimized fermentation conditions. As a consequence, metabolically engineered E. coli strain produced 847.97 mg/L of 3-phenypropanol at 24 h using glucose-glycerol mixture as co-carbon source. Conclusions We successfully developed an artificial 3-phenylpropanol pathway based on retrobiosynthesis approach, and highest titer of 3-phenylpropanol was achieved in E. coli via systems metabolic engineering strategies including enzyme sources variety, chromosome engineering, metabolic strength balancing and fermentation optimization. This work provides an engineered strain with industrial potential for production of 3-phenylpropanol, and the strategies applied here could be practical for bioengineers to design and reconstruct the microbial cell factory for high valuable chemicals.

De Novo Metabolic Engineering and the Promise of Synthetic DNA

2010 ◽  
pp. 101-131 ◽  
Author(s):  
Daniel Klein-Marcuschamer ◽  
Vikramaditya G. Yadav ◽  
Adel Ghaderi ◽  
Gregory N. Stephanopoulos
Keyword(s):  
Metabolic Engineering ◽  
De Novo ◽  
Synthetic Dna

scholarly journals Metabolic engineering of Escherichia coli BL21 (DE3) for de novo production of l-DOPA from d-glucose

2019 ◽  
Vol 18 (1) ◽  
Author(s):  
Eric Fordjour ◽  
Frederick Komla Adipah ◽  
Shenghu Zhou ◽  
Guocheng Du ◽  
Jingwen Zhou
Keyword(s):  
Escherichia Coli ◽  
Metabolic Engineering ◽  
De Novo ◽  
Escherichia Coli Bl21

scholarly journals Draft Genome Sequences of Itaconate-Producing Ustilaginaceae

2016 ◽  
Vol 4 (6) ◽  
Author(s):  
Elena Geiser ◽  
Florian Ludwig ◽  
Thiemo Zambanini ◽  
Nick Wierckx ◽  
Lars M. Blank
Keyword(s):  
Metabolic Engineering ◽  
Genome Sequencing ◽  
De Novo ◽  
Draft Genome ◽  
Essential Genes ◽  
Smut Fungi ◽  
Genome Sequences ◽  
The Family

Some smut fungi of the family Ustilaginaceae produce itaconate from glucose. De novo genome sequencing of nine itaconate-producing Ustilaginaceae revealed genome sizes between 19 and 25 Mbp. Comparison to the itaconate cluster of U. maydis MB215 revealed all essential genes for itaconate production contributing to metabolic engineering for improving itaconate production.

scholarly journals Redistribution of Carbon Flux toward 2,3-Butanediol Production in Klebsiella pneumoniae by Metabolic Engineering

PLoS ONE ◽  
2014 ◽  
Vol 9 (10) ◽  
pp. e105322 ◽  
Author(s):  
Borim Kim ◽  
Soojin Lee ◽  
Daun Jeong ◽  
Jeongmo Yang ◽  
Min-Kyu Oh ◽  
...  
Keyword(s):  
Metabolic Engineering ◽  
Klebsiella Pneumoniae ◽  
Carbon Flux

scholarly journals Application of Random Mutagenesis and Synthetic FadR Promoter for de novo Production of ω-Hydroxy Fatty Acid in Yarrowia lipolytica

2021 ◽  
Vol 9 ◽  
Author(s):  
Beom Gi Park ◽  
Junyeob Kim ◽  
Eun-Jung Kim ◽  
Yechan Kim ◽  
Joonwon Kim ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Metabolic Engineering ◽  
Palmitic Acid ◽  
Yarrowia Lipolytica ◽  
Cell Sorting ◽  
De Novo ◽  
Random Mutagenesis ◽  
Oleaginous Yeasts ◽  
Synthetic Promoters ◽  
Base Strain

As a means to develop oleaginous biorefinery, Yarrowia lipolytica was utilized to produce ω-hydroxy palmitic acid from glucose using evolutionary metabolic engineering and synthetic FadR promoters for cytochrome P450 (CYP) expression. First, a base strain was constructed to produce free fatty acids (FFAs) from glucose using metabolic engineering strategies. Subsequently, through ethyl methanesulfonate (EMS)-induced random mutagenesis and fluorescence-activated cell sorting (FACS) screening, improved FFA overproducers were screened. Additionally, synthetic promoters containing bacterial FadR binding sequences for CYP expression were designed to respond to the surge of the concentration of FFAs to activate the ω-hydroxylating pathway, resulting in increased transcriptional activity by 14 times from the third day of culture compared to the first day. Then, endogenous alk5 was screened and expressed using the synthetic FadR promoter in the developed strain for the production of ω-hydroxy palmitic acid. By implementing the synthetic FadR promoter, cell growth and production phases could be efficiently decoupled. Finally, in batch fermentation, we demonstrated de novo production of 160 mg/L of ω-hydroxy palmitic acid using FmeN3-TR1-alk5 in nitrogen-limited media. This study presents an excellent example of the production of ω-hydroxy fatty acids using synthetic promoters with bacterial transcriptional regulator (i.e., FadR) binding sequences in oleaginous yeasts.

scholarly journals One Health Genomic Study of Human and Animal Klebsiella pneumoniae Isolated at Diagnostic Laboratories on a Small Caribbean Island

Antibiotics ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 42
Author(s):  
Patrick Butaye ◽  
Marc Stegger ◽  
Arshnee Moodley ◽  
Peter Damborg ◽  
Andrea Williams ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
One Health ◽  
De Novo ◽  
Large Population ◽  
Vervet Monkeys ◽  
Nucleotide Polymorphisms ◽  
Caribbean Island ◽  
Indirect Contact ◽  
Genomic Study ◽  
Antimicrobial Resistance Gene

Klebsiella pneumoniae causes a variety of infections in both humans and animals. In this study, we characterised the genomes of human and animal isolates from two diagnostic laboratories on St. Kitts, a small Caribbean island inhabited by a large population of vervet monkeys. In view of the increased chances of direct or indirect contact with humans and other animal species, we used the One Health approach to assess transmission of K. pneumoniae across host species by sequencing 82 presumptive K. pneumoniae clinical isolates from humans (n = 51), vervets (n = 21), horses (n = 5), dogs (n = 4) and a cat (n = 1). Whole genome sequencing (WGS) was carried out using Illumina technology. De novo assembly was performed in CLC Genomics Workbench v.11.0. Single nucleotide polymorphisms were detected using NASP followed by phylogenetic analysis using IQ-TREE. Virulence and antimicrobial resistance gene contents were analysed using the Kleborate and CGE pipelines. WGS-based analysis showed that 72 isolates were K. pneumoniae sensu stricto and five K. quasipneumoniae and five K. variicola. K. pneumoniae isolates belonged to 35 sequence types (ST), three of which were occasionally shared between humans and animals: ST23, ST37 and ST307. The ST23 strains from vervets formed a separate cluster amongst publicly available sequenced ST23 strains, indicating the presence of a specific vervet sublineage. Animal strains harbored fewer resistance genes and displayed distinct virulence traits that appeared to be host-specific in vervet isolates. Our results show that K. pneumoniae infections on this Caribbean island are usually caused by host-specific lineages.

scholarly journals Epidemiology of paediatric gastrointestinal colonisation by extended spectrum cephalosporin-resistant Escherichia coli and Klebsiella pneumoniae isolates in north-west Cambodia

2017 ◽  
Author(s):  
JJ van Aartsen ◽  
CE Moore ◽  
CM Parry ◽  
P Turner ◽  
N Phot ◽  
...  
Keyword(s):  
Risk Factors ◽  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Resistance Genes ◽  
Genetic Relatedness ◽  
De Novo ◽  
Intestinal Parasites ◽  
E Coli ◽  
Extended Spectrum ◽  
Independent Risk Factors

ABSTRACTExtended-spectrum cephalosporin resistance (ESC-R) in Escherichia coli and Klebsiella pneumoniae is a healthcare threat; high gastrointestinal carriage rates are reported from South-east Asia. Colonisation prevalence data in Cambodia are lacking. We determined gastrointestinal colonisation prevalence of ESC-resistant E. coli (ESC-R-EC) and K. pneumoniae (ESC-R-KP) in Cambodian children/adolescents and associated risk factors; characterised relevant resistance genes, their genetic contexts, and the genetic relatedness of ESC-R strains using whole genome sequencing (WGS). Faeces and questionnaire data were obtained from individuals <16 years in northwestern Cambodia, 2012. WGS of cultured ESC-R-EC/KP was performed (Illumina). Maximum likelihood phylogenies were used to characterise relatedness of isolates; ESC-R-associated resistance genes and their genetic contexts were identified from de novo assemblies using BLASTn and automated/manual annotation. 82/148 (55%) of children/adolescents were ESC-R-EC/KP colonised; 12/148 (8%) were co-colonised with both species. Independent risk factors for colonisation were hospitalisation (OR: 3.12, 95%, CI [1.52-6.38]) and intestinal parasites (OR: 3.11 [1.29-7.51]); school attendance conferred decreased risk (OR: 0.44 [0.21-0.92]. ESC-R strains were diverse; the commonest ESC-R mechanisms were blaCTX-M 1 and 9 sub-family variants. Structures flanking these genes were highly variable, and for blaCTX-M-15,-55and-27, frequently involved IS26. Chromosomal blaCTX-M integration was common in E. coli. Gastrointestinal ESC-R-EC/KP colonisation is widespread in Cambodian children/adolescents; hospital admission and intestinal parasites are independent risk factors. The genetic contexts of blaCTX-M are highly mosaic, consistent with rapid horizontal exchange. Chromosomal integration of blaCTX-M may result in stable propagation in these community-associated pathogens.

Sign in / Sign up

Export Citation Format

Share Document