• Journal of Microbiology and Biotechnology
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• 제30권10호
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• pp.1592-1596
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• 2020

The aerobic growth and metabolic performance of Escherichia coli strains BL21 and W3110 were studied when the Vitreoscilla hemoglobin (VHb) was constitutively expressed in the chromosome. When VHb was expressed, acetate production decreased in both strains and was nearly eliminated in BL21. Transcriptional levels of the glyoxylate shunt genes decreased in both strains when VHb was expressed. However, higher transcription of the α-ketoglutarate dehydrogenase genes were observed for W3110, while for BL21 transcription levels decreased. VHb expression reduced the transcription of the cytochrome bo₃ genes only in BL21. These results are useful for better selecting a production host.

• Biotechnology and Bioprocess Engineering:BBE
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• 제9권4호
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• pp.274-277
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• 2004

Production of green fluorescent protein (GFP) as a model foreign protein using different culture scales under co-expression of Vitreoscilla hemoglobin (VHb) in the industrial Escherichia coli strain W3110 (a K12 derivative), was examined. It was found that the VHb co-expressing W3110, exhibited an exceptional and sustained production ability during cell cultures using different scales, while the VHb non-expressing strain showed variable production levels. This high and sustained production ability indicates that the VHb co-expressing E. coli W3110, could be successfully employed for practical large-scale production cultures without the need for serious consideration of scale-up problems.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.754-757
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• 2001

Expression of the vhb gene encoding bacterial hemoglobin (VHb) from Vitreoscilla has been used to improve recombinant cell growth and enhance product formation under microaerobic conditions because of its ability to enhance oxygen use. We coexpressed GFP and VHb in Escherichia coli BL21 and W3110, and compared with GFP control which was not expressed VHb. We used nar oxygen-dependent inducible promoter for VHb expression. The GFP amounts in E. coli expressed VHb was about five fold higher than in the control Fluorescence intensity was increased about two fold.

• ALGAE
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• 제17권3호
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• pp.195-199
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• 2002

We investigated the association of Chlorella vulgaris and E. coli W9110 in removal of total organic carbon with the lab-scaled continuous river water flow system (CRWFS). Artificial wastewater was applied at two levels of organic carbon concentration; 1,335 $mg{\cdot}l^{-1}$ in the treatment (T)-1 and 267 $mg{\cdot}l^{-1}$ in T-2. The highest densities of C. vulgaris were $8.3{\times10^6\;cells{\cdot}ml^{-1}$ in T-1 and $6.9{\times}10^6\;cells{\cdot}ml^{-1}$ in T-2. The maximum densities of E. coli W3110 were $2.0{\times}10^8$ clony forming unit (CFU)${\cdot}ml^{-1}$ in T-1 and $3.9{\times}10^8\;CFU{\cdot}ml^{-1}$ in T-2. The densities increased during the first 11 days in T-q and 4 days in T-2, and decreased rapidly till 35th day, then increased slightly afterwards. This trend was prominent in T-2. It was inplied that wider range of nutrients was required in the growth of heterotrophic bacteria in T-2 than in T-1. The algal biomass should be increased effectively for the successful removal of organic carbon.

• 한국환경과학회지
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• 제12권8호
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• pp.847-852
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• 2003

It was examined carefully that the bacterial die-off between Chlorella vulgaris and E. coli. W3110 was tested through adding TOC (total organic carbon) with the lab-scaled continuous river water flow system (CRWFS). Artificial synthetic wastewater was applied at two levels of organic carbon concentration; 1,335 mg/l in treatment type 1 and 267 mg/l in type 2. In both types, the population densities of Chlorella vulgaris were similar in a maximum 8.25 ${\times}$ 10$\^$6/ cells/ml (type 1) and 6.925 ${\times}$ 10$\^$6/ cells/ml (type 2). The maximum densities of E. coli. W3110 were 2.0 ${\times}$ 10$\^$8/ colony forming unit (CFU)/ml in type 1 and 3.9 ${\times}$ 10$\^$8/ CFU/ml in type 2. The densities increased for 11 days in type 1 and 4 days in type 2, then decreased rapidly till the 35th day, then slightly increased again. This trend was prominent in type 2. It implied that a wider range of nutrients was required in the growth of heterotrophic bacteria in type 2 than in type 1. We could not expect successful bacterial die-off if the wastewater retention time was not furnished sufficiently.

• 한국미생물·생명공학회지
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• 제19권6호
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• pp.583-587
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• 1991

대장균 W3110으로부터 NTG 및 UV를 사용하여 여러 단계의 돌연변이 실험을 거치면서 스레오닌 고생산균주인 대장균 TF427를 선별하였다. 선별된 변이주는 스레오닌 유사체인 AHV 내성, 메치오닌 및 이소루이신 요구성을 특징으로 한다. 5-L 발효조 실험에서 44시간 발효하였을 때 46.5g/l의 스레오닌이 생산되었다. 효모분석에 의하면, TF427의 아스파토키나아제 I의 활성은 스레오닌에 의해 저해받지 않았으며, 이 효소의 합성은 스레오닌과 이소루이신에 의해 억제를 받지 않았다.

• Journal of Microbiology and Biotechnology
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• 제26권8호
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• pp.1452-1456
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• 2016

Overexpression of the genes encoding phosphoeneolpyruvate carboxykinase (pckA) and NAD-dependent malic enzyme (maeA) facilitates higher intracellular ATP and NAD(P)H concentrations, respectively, under aerobic conditions in Escherichia coli. To verify a hypothesis that higher intracellular energy reserves might contribute to H₂ fermentation, wild-type E. coli strains overexpressing pckA and maeA were cultured under anaerobic conditions in a glucose minimal medium. Overexpression of pckA and maeA enabled E. coli to produce 3-times and 4-times greater H₂ (193 and 284 nmol, respectively) than the wild type (66 nmol H₂). The pckA and maeA genes were further overexpressed in a hydrogenase-3-enhanced E. coli strain. The hydrogenase-3-enhanced strain (W3110+fhlA) produced 322 nmol H₂, whereas the ATP-enhanced strain (W3110+fhlA+pckA) produced 50% increased H₂ (443 nmol). Total H₂ in the NAD(P)H-enhanced strain (W3110+fhlA+maeA) was similar to that in the control strain at 319 nmol H₂. Possible explanations for the contribution of the increased cellular energy reserves to the enhanced hydrogen fermentation observed are discussed based on the viewpoint of metabolic engineering strategy.

• Journal of Microbiology and Biotechnology
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• 제21권4호
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• pp.374-378
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• 2011

To find alternative genetic resources for D-serine dehydratase (E.C. 4.3.1.18, dsdA) mediating the deamination of D-serine into pyruvate, metagenomic libraries were screened. The chromosomal dsdA gene of a wild-type Escherichia coli W3110 strain was disrupted by inserting the tetracycline resistance gene (tet), using double-crossover, for use as a screening host. The W3110 dsdA::tet strain was not able to grow in a medium containing D-serine as a sole carbon source, whereas wild-type W3110 and the complement W3110 dsdA::tet strain containing a dsdA-expression plasmid were able to grow. After introducing metagenome libraries into the screening host, a strain containing a 40-kb DNA fragment obtained from the metagenomic souce derived from a compost was selected based on its capability to grow on the agar plate containing D-serine as a sole carbon source. For identification of the genetic resource responsible for the D-serine degrading capability, transposon-${\mu}$ was randomly inserted into the 40-kb metagenome. Two strains that had lost their D-serine degrading ability were negatively selected, and the two 6-kb contigs responsible for the D-serine degrading capability were sequenced and deposited (GenBank code: HQ829474.1 and HQ829475.1). Therefore, new alternative genetic resources for D-serine dehydratase was found from the metagenomic resource, and the corresponding ORFs are discussed.

• KSBB Journal
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• 제29권4호
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• pp.244-249
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• 2014

Biosynthesis pathway of medium-chain-length (MCL) polyhydroxyalkanoates (PHA) from fatty acid ${\beta}$-oxidation pathway was constructed in recombinant Escherichia coli by introducing the Pseudomonas sp. 61-3 PHA synthase gene (phaC2) and the maoC genes from Pseudomonas putida, Sinorhizobium meliloti, and Ralstonia eutropha. The metabolic link between fatty acid ${\beta}$-oxidation pathway and PHA biosynthesis pathway was constructed by MaoC, which is homologous to P. aeruginosa (R)-specific enoyl-CoA hydratase (PhaJ1). When the E. coli W3110 strains expressing the phaC2 gene and one of the maoC genes from P. putida, Sinorhizobium meliloti, and Ralstonia eutropha were cultured in LB medium containing 2 g/L of sodium decanoate as a carbon source, MCL-PHA that mainly consists of 3-hydroxyhexanoate (3HHx), 3-hydroxyoctanoate (3HO) and 3-hydroxydecanoate (3HD), was produced. The monomer composition of PHA and PHA contents varied depending on MaoC employed for the production of PHA. The highest PHA content of 18.7 wt% was achieved in recombinant E. coli W3110 expressing the phaC2 gene and the P. putida maoC gene. These results suggest that MCL-PHA biosynthesis pathway can be constructed in recombinant E. coli strains from the b-oxidation pathway by employing MaoC able to supply (R)-3-hydroxyacyl-CoA, the substrate of PHA synthase.

• Biotechnology and Bioprocess Engineering:BBE
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• 제11권2호
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• pp.164-167
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• 2006

The influence of temperature and agitation on the growth of Escherichia coli expressing hepatitis B core antigen (HBcAg) in stirred tank bioreactor were investigated. The highest specific growth rate for E. coli$(0.844 h^{-1})$ was achieved at a temperature of $37^{\circ}C$ and an agitation speed of 250 rpm. The activation energy for the growth of the E. coli strain W3110lQ in the stirred tank bioreactor was estimated to be 11 kcal/mol. The highest protein yield was achieved at a temperature of $44^{\circ}C$ and an agitation speed of 250 rpm. The relative protein concentration at $44^{\circ}C$ is 30 and 6% higher compared to that at 30 and $37^{\circ}C$, respectively.