Semirational Approach for Ultrahigh Poly(3-hydroxybutyrate) Accumulation in <i>Escherichia coli</i> by Combining One-Step Library Construction and High-Throughput Screening

Li, Teng; Ye, Jianwen; Shen, R.; Zong, Yeqing; Zhao, Xuejin; Lou, Chunbo; Chen, Guoqiang

doi:10.1021/acssynbio.6b00083

Cited by 70 publications

(34 citation statements)

References 36 publications

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“…In this study, nine promoters were chosen from the 96-promoter library and were named P1-P9 (Supporting Information Table S3). They were named H-PC, H3, and H36 (T. Li, Ye, et al, 2016). They were named H-PC, H3, and H36 (T. Li, Ye, et al, 2016).…”

Section: Construction Of Pporin-phacab Cn Library For H Campaniensmentioning

confidence: 99%

“…The pretreatment process of cells for staining was modified from a previously reported protocol (J. H. Lee et al, 2013;T. Li , Ye, et al, 2016).…”

Section: Cells Staining With Boron-dipyrromethene (Bodipy) Fluorescmentioning

confidence: 99%

“…Zhao et al, 2017). Nine promoters P porin of various strengths were combined with the three RBS optimized phaCAB cn operons including H36, H3 and H-PC (T. Li, Ye, et al, 2016), respectively. This study identified another 43 bp core region of P porin in H. campaniensis LS21, based on which nine promoters were screened with strength variations over 1700 folds (Table.…”

Section: Enhanced Phb Synthesis By Overexpressing Heterologous Phacmentioning

confidence: 99%

“…The nine promoters containing the highest induction strength of more than 300 folds and good linear correlation, were further combined with three different ribosome binding site (RBS)-optimized PHA synthesis operons, namely, pha-CAB cn operons (phaCAB from Cupriavidus necator). They were named H-PC, H3, and H36 (T. Li, Ye, et al, 2016). All the plasmids were constructed based on the pBMCS1-OriC backbone (Yue et al, 2014) using Gibson assembly method (Gibson et al, 2009).…”

Section: Construction Of Pporin-phacab Cn Library For H Campaniensmentioning

confidence: 99%

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Engineering self‐flocculating Halomonas campaniensis for wastewaterless open and continuous fermentation

Chen

Qiao

Shuai

et al. 2019

Biotech & Bioengineering

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Halomonas has been developed as a platform for the next generation industrial biotechnology allowing open and nonsterile growth without microbial contamination under a high-salt concentration and alkali pH. To reduce downstream cost associated with continuous centrifugation and salt containing wastewater treatment, Halomonas campaniensis strain LS21 was engineered to become self-flocculating by knocking out an etf operon encoding two subunits of an electron transferring flavoprotein in the predicted electron transfer chain. Self-flocculation could be attributed to the decrease of the surface charge and increase of the cellular hydrophobicity resulted from deleted etf. A wastewaterless fermentation strategy based on the self-flocculating H. campaniensis was developed for growth and the production of poly-3-hydroxybutyrate (PHB) as an example. Most microbial cells flocculated and precipitated to the bottom of the bioreactor within 1 min after stopping the aeration and agitation. The supernatant can be used again without sterilization or inoculation for the growth of the next batch after collecting the precipitated cell mass. The wastewaterless process was conducted for four runs without generating wastewater. PHB accumulation by the self-flocculent strain was enhanced via promoter and ribosome binding site optimizations, the productivities of cell dry weight and PHB were increased from 0.45 and 0.18 g·L −1 ·hr −1 for the batch process compared to 0.82 and 0.33 g·L −1 ·hr −1 for the wastewaterless continuous process, respectively. This has clearly demonstrated the advantages of the wastewaterless process in that it not only reduces wastewater but also increases cell growth and product formation efficiency in a given period of time. K E Y W O R D S continuous process, etf, flocculation, Halomonas campaniensis, next generation industry biotechnology, NGIB, PHB

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Section: Construction Of Pporin-phacab Cn Library For H Campaniensmentioning

confidence: 99%

“…The pretreatment process of cells for staining was modified from a previously reported protocol (J. H. Lee et al, 2013;T. Li , Ye, et al, 2016).…”

Section: Cells Staining With Boron-dipyrromethene (Bodipy) Fluorescmentioning

confidence: 99%

Section: Enhanced Phb Synthesis By Overexpressing Heterologous Phacmentioning

confidence: 99%

Section: Construction Of Pporin-phacab Cn Library For H Campaniensmentioning

confidence: 99%

See 2 more Smart Citations

Engineering self‐flocculating Halomonas campaniensis for wastewaterless open and continuous fermentation

Chen

Qiao

Shuai

et al. 2019

Biotech & Bioengineering

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“…Therefore, fine-tuning expression with a combination of suitable promoters and ribosome binding sites, and dynamic regulation of key enzymes are useful for enhancing the production from synthetic pathways [57][58][59][60][61][62]. The primary consideration is to tune the expression levels of multiple enzymes of the synthetic pathway to achieve a good balance.…”

Section: Compatibility Within Synthetic Pathwaysmentioning

confidence: 99%

Advances in bio‐based production of dicarboxylic acids longer than C4

Qian

Zhong

2018

Engineering in Life Sciences

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Growing concerns of environmental pollution and fossil resource shortage are major driving forces for bio‐based production of chemicals traditionally from petrochemical industry. Dicarboxylic acids (DCAs) are important platform chemicals with large market and wide applications, and here the recent advances in bio‐based production of straight‐chain DCAs longer than C4 from biological approaches, especially by synthetic biology, are reviewed. A couple of pathways were recently designed and demonstrated for producing DCAs, even those ranging from C5 to C15, by employing respective starting units, extending units, and appropriate enzymes. Furthermore, in order to achieve higher production of DCAs, enormous efforts were made in engineering microbial hosts that harbored the biosynthetic pathways and in improving properties of biocatalytic elements to enhance metabolic fluxes toward target DCAs. Here we summarize and discuss the current advantages and limitations of related pathways, and also provide perspectives on synthetic pathway design and optimization for hyper‐production of DCAs.

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Microbial Polyhydroxyalkanoates and Nonnatural Polyesters

et al. 2020

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Microorganisms produce diverse polymers for various purposes such as storing genetic information, energy, and reducing power, and serving as structural materials and scaffolds. Among these polymers, polyhydroxyalkanoates (PHAs) are microbial polyesters synthesized and accumulated intracellularly as a storage material of carbon, energy, and reducing power under unfavorable growth conditions in the presence of excess carbon source. PHAs have attracted considerable attention for their wide range of applications in industrial and medical fields. Since the first discovery of PHA accumulating bacteria about 100 years ago, remarkable advances have been made in the understanding of PHA biosynthesis and metabolic engineering of microorganisms toward developing efficient PHA producers. Recently, nonnatural polyesters have also been synthesized by metabolically engineered microorganisms, which opened a new avenue toward sustainable production of more diverse plastics. Herein, the current state of PHAs and nonnatural polyesters is reviewed, covering mechanisms of microbial polyester biosynthesis, metabolic pathways, and enzymes involved in biosynthesis of short‐chain‐length PHAs, medium‐chain‐length PHAs, and nonnatural polyesters, especially 2‐hydroxyacid‐containing polyesters, metabolic engineering strategies to produce novel polymers and enhance production capabilities and fermentation, and downstream processing strategies for cost‐effective production of these microbial polyesters. In addition, the applications of PHAs and prospects are discussed.

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Semirational Approach for Ultrahigh Poly(3-hydroxybutyrate) Accumulation in Escherichia coli by Combining One-Step Library Construction and High-Throughput Screening

Cited by 70 publications

References 36 publications

Engineering self‐flocculating Halomonas campaniensis for wastewaterless open and continuous fermentation

Engineering self‐flocculating Halomonas campaniensis for wastewaterless open and continuous fermentation

Advances in bio‐based production of dicarboxylic acids longer than C4

Microbial Polyhydroxyalkanoates and Nonnatural Polyesters

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