Biosynthesis of polyhydroxyalkanoates from sucrose by metabolically engineered Escherichia coli strains

Sohn, Yu Jung; Kim, Hee Taek; Baritugo, Kei Anne; Song, Hyun Keun; Ryu, Mi Hee; Kang, Kyoung Hee; Jo, Seo Young; Kim, Hoyong; Kim, You Jin; Choi, Jong Il; Park, Su Kyeong; Joo, Jeong Chan; Park, Si Jae

doi:10.1016/j.ijbiomac.2020.01.254

Cited by 33 publications

(17 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Although Pseudomonas putida is the most studied PHA-producing strain, species including P. putida and its variants have difficulty using sucrose directly [ 25 ]. Because sugarcane-based feedstocks and waste fructose syrup are cheap and abundant sources for PHA production, several efforts have been undertaken to metabolically engineer strains to fully uptake sucrose, pretreat feedstock to degrade sucrose to glucose and fructose, or discover strains that are able to utilize sucrose to produce PHA [ 25 , 26 , 27 ].…”

Section: Introductionmentioning

confidence: 99%

Fructose-Based Production of Short-Chain-Length and Medium-Chain-Length Polyhydroxyalkanoate Copolymer by Arctic Pseudomonas sp. B14-6

et al. 2021

View full text Add to dashboard Cite

Arctic bacteria employ various mechanisms to survive harsh conditions, one of which is to accumulate carbon and energy inside the cell in the form of polyhydroxyalkanoate (PHA). Whole-genome sequencing of a new Arctic soil bacterium Pseudomonas sp. B14-6 revealed two PHA-production-related gene clusters containing four PHA synthase genes (phaC). Pseudomonas sp. B14-6 produced poly(6% 3-hydroxybutyrate-co-94% 3-hydroxyalkanoate) from various carbon sources, containing short-chain-length PHA (scl-PHA) and medium-chain-length PHA (mcl-PHA) composed of various monomers analyzed by GC-MS, such as 3-hydroxybutyrate, 3-hydroxyhexanoate, 3-hydroxyoctanoate, 3-hydroxydecanoate, 3-hydroxydodecenoic acid, 3-hydroxydodecanoic acid, and 3-hydroxytetradecanoic acid. By optimizing the PHA production media, we achieved 34.6% PHA content using 5% fructose, and 23.7% PHA content using 5% fructose syrup. Differential scanning calorimetry of the scl-co-mcl PHA determined a glass transition temperature (Tg) of 15.3 °C, melting temperature of 112.8 °C, crystallization temperature of 86.8 °C, and 3.82% crystallinity. In addition, gel permeation chromatography revealed a number average molecular weight of 3.6 × 104, weight average molecular weight of 9.1 × 104, and polydispersity index value of 2.5. Overall, the novel Pseudomonas sp. B14-6 produced a polymer with high medium-chain-length content, low Tg, and low crystallinity, indicating its potential use in medical applications.

show abstract

Section: Introductionmentioning

confidence: 99%

Fructose-Based Production of Short-Chain-Length and Medium-Chain-Length Polyhydroxyalkanoate Copolymer by Arctic Pseudomonas sp. B14-6

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Hence, various bio-based production methods using renewable feedstocks have recently been developed [1,2]. To address the growing demand for the production of value-added products, such as platform chemicals and biopolymers from renewable resources, several engineered microorganisms have been developed [3][4][5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

Effect of DR1558, a Deinococcus radiodurans response regulator, on the production of GABA in the recombinant Escherichia coli under low pH conditions

et al. 2020

Self Cite

View full text Add to dashboard Cite

Background: Gamma aminobutyric acid (GABA) is an important platform chemical, which has been used as a food additive and drug. Additionally, GABA is a precursor of 2-pyrrolidone, which is used in nylon synthesis. GABA is usually synthesized from glutamate in a reaction catalyzed by glutamate decarboxylase (GAD). Currently, there are several reports on GABA production from monosodium glutamate (MSG) or glucose using engineered microbes. However, the optimal pH for GAD activity is 4, which is the limiting factor for the efficient microbial fermentative production of GABA as fermentations are performed at pH 7. Recently, DR1558, a response regulator in the two-component signal transduction system was identified in Deinococcus radiodurans. DR1558 is reported to confer cellular robustness to cells by binding the promoter regions of genes via DNA-binding domains or by binding to the effector molecules, which enable the microorganisms to survive in various environmental stress conditions, such as oxidative stress, high osmotic shock, and low pH. Results: In this study, the effect of DR1558 in enhancing GABA production was examined using two different strategies: whole-cell bioconversion of GABA from MSG and direct fermentative production of GABA from glucose under acidic culture conditions. In the whole-cell bioconversion, GABA produced by E. coli expressing GadBC and DR1558 (6.52 g/L GABA from 13 g/L MSG•H 2 O) in shake flask culture at pH 4.5 was 2.2-fold higher than that by E. coli expressing only GadBC (2.97 g/L of GABA from 13 g/L MSG•H 2 O). In direct fermentative production of GABA from glucose, E. coli ∆gabT expressing isocitrate dehydrogenase (IcdA), glutamate dehydrogenase (GdhA), GadBC, and DR1558 produced 1.7-fold higher GABA (2.8 g/L of GABA from 30 g/L glucose) than E. coli ∆gabT expressing IcdA, GdhA, and GadBC (1.6 g/L of GABA from 30 g/L glucose) in shake flask culture at an initial pH 7.0. The transcriptional analysis of E. coli revealed that DR1558 conferred acid resistance to E. coli during GABA production. The fed-batch fermentation of E. coli expressing IcdA, GdhA, GadBC, and DR1558 performed at pH 5.0 resulted in the final GABA titer of 6.16 g/L by consuming 116.82 g/L of glucose in 38 h. Conclusion: This is the first report to demonstrate GABA production by acidic fermentation and to provide an engineering strategy for conferring acid resistance to the recombinant E. coli for GABA production.

show abstract

“…Synthesis of PHB via recombinant E. coli strain has received extensive attention, and numerous studies attempted to reduce the production cost of PHB to promote its commercialization [ 20 , 21 , 22 , 23 , 24 ]. For example, by optimizing the metabolic pathways of PHB, an engineered E. coli can use crude glycerol as the sole carbon source to produce PHB and reach a PHB content of 65% dry cell weight after fermentation [ 20 ].…”

Section: Introductionmentioning

confidence: 99%

“…For example, by optimizing the metabolic pathways of PHB, an engineered E. coli can use crude glycerol as the sole carbon source to produce PHB and reach a PHB content of 65% dry cell weight after fermentation [ 20 ]. Moreover, construction of the sucrose utilization pathway in an engineered E. coli XL1-Blue strain resulted in the production of 38 wt.% PHB with 20 g/L sucrose as the sole carbon source [ 21 ]. In addition to expanding the repertoire of the widely available and low-cost substrates such as sucrose, xylose, and glycerol, many studies also aimed at reducing the energy consumption in the fermentation processes, by utilizing hypoxic conditions that are more conducive to the accumulation of PHB [ 22 , 23 , 24 ].…”

Section: Introductionmentioning

confidence: 99%

“…A sucrose utilization pathway was constructed in E. coli by expressing the β-fructofuranosidase (SacC) from Mannheimia succiniciproducens MBEL 55E [ 29 ]. By introducing the sacC gene, E. coli could convert sucrose into glucose and fructose [ 21 ]. In this way, the recombinant strain becomes a cost-effective and sustainable host to produce PHB and other materials.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Biosynthesis of Poly-(3-hydroxybutyrate) under the Control of an Anaerobically Induced Promoter by Recombinant Escherichia coli from Sucrose

Zhou

Pei

et al. 2022

Molecules

View full text Add to dashboard Cite

Poly-(3-hydroxybutyrate) (PHB) is a polyester with biodegradable and biocompatible characteristics and has many potential applications. To reduce the raw material costs and microbial energy consumption during PHB production, cheaper carbon sources such as sucrose were evaluated for the synthesis of PHB under anaerobic conditions. In this study, metabolic network analysis was conducted to construct an optimized pathway for PHB production using sucrose as the sole carbon source and to guide the gene knockout to reduce the generation of mixed acid byproducts. The plasmid pMCS-sacC was constructed to utilize sucrose as a sole carbon source, and the cascaded promoter P3nirB was used to enhance PHB synthesis under anaerobic conditions. The mixed acid fermentation pathway was knocked out in Escherichia coli S17-1 to reduce the synthesis of byproducts. As a result, PHB yield was improved to 80% in 6.21 g/L cell dry weight by the resulted recombinant Escherichia coli in a 5 L bed fermentation, using sucrose as the sole carbon source under anaerobic conditions. As a result, the production costs of PHB will be significantly reduced.

show abstract

Biosynthesis of polyhydroxyalkanoates from sucrose by metabolically engineered Escherichia coli strains

Cited by 33 publications

References 34 publications

Fructose-Based Production of Short-Chain-Length and Medium-Chain-Length Polyhydroxyalkanoate Copolymer by Arctic Pseudomonas sp. B14-6

Fructose-Based Production of Short-Chain-Length and Medium-Chain-Length Polyhydroxyalkanoate Copolymer by Arctic Pseudomonas sp. B14-6

Effect of DR1558, a Deinococcus radiodurans response regulator, on the production of GABA in the recombinant Escherichia coli under low pH conditions

Biosynthesis of Poly-(3-hydroxybutyrate) under the Control of an Anaerobically Induced Promoter by Recombinant Escherichia coli from Sucrose

Contact Info

Product

Resources

About