Molecular Weight Change of Polyhydroxyalkanoate (PHA) Caused by the PhaC Subunit of PHA Synthase from Bacillus cereus YB-4 in Recombinant Escherichia coli

Tomizawa, Satoshi; Hyakutake, Manami; Saito, Yuta; Agus, Jumiarti; Mizuno, Kouhei; Abe, Hideki; Tsuge, Takeharu

doi:10.1021/bm2004687

Cited by 49 publications

(46 citation statements)

References 26 publications

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“…The most important genes that involves in PHB production in Bacillus strains were phaA and phaR, phaB, phaC [3,23]. On the other hand, Valappil et al [25] and Tomizawa et al [14] proved that in Bacillus genus, two proteins, PhaC, the normal PHA synthase enzyme, and PhaR have been proven to be essential for the PHA synthase activity. The PHA synthases was exist in two forms; an active form in PHAaccumulating cells and an inactive form in non-accumulating cells, where PhaC is more susceptible to degradation in cell [18].…”

Section: Discussionmentioning

confidence: 99%

“…PHA biosynthetic genes from different species were cloned for mass production of PHAs for commercial use [6,[11][12][13]. Tomizawa et al [14] studied the expression of PHA synthases from Bacillus megaterium NBRC15308 (PhaRCBm), B. cereus YB-4 (PhaRCYB4) and hybrids (PhaRBmCYB4 and PhaRYB4CBm) in Escherichia coli JM109 to characterize the molecular weight of the synthesized poly (3-hydroxybutyrate) [P (3HB)].…”

Section: Introductionmentioning

confidence: 99%

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Isolation, Cloning and Sequencing of Poly 3-Hydroxybutyrate Synthesis Genes from Local Strain of Bacillus cereus mm7 and Expression them in E. coli

Rabey¹

2017

JIG

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Poly 3-hydroxybutyrate (PHB) is the most known degradable biopolymers, produced by some genera of bacteria under unfavorable growth conditions. Isolation, cloning, and expression of PHB genes from PHB producing Bacillus cereus was achieved. A highest Poly 3-hydroxybutyrate producer bacterium was selected and identified as Bacillus cereus MM7, based on morphological and physiological characters in addition to 16S rRNA gene sequences. The four genes of Bacillus cereus MM7 that involved in Poly 3-hydroxybutyrate production were successfully amplified using specific primers which were designed depending on the sequence alignment of the known PHA biosynthetic genes from Bacillus cereus, cloned in E. coli DH5α and completely sequenced. Moreover, the expression of the four genes responsible for PHA biosynthetic in E. coli XL1-Blue and E coli MJ 109 was assayed by 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis. In the current study Bacillus cereus was able to synthesize PHB. PHB genes were isolated, cloned and expressed in E. coli DH5α. The PHB encoding proteins were analyzed in SDS-PAGE and the PHB corresponding protein bands were analyzed using mass spectrometry to nominate their amino acid sequence. Keywords: Pre cultureThe pre culture was prepared in 250 ml Erlenmeyer flasks containing 50 ml of nutrient broth [16]. Each flask was inoculated with a loop of the tested cultures and incubated in a 130 rpm speed rotary shaker at 30˚C for 24h.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Isolation, Cloning and Sequencing of Poly 3-Hydroxybutyrate Synthesis Genes from Local Strain of Bacillus cereus mm7 and Expression them in E. coli

Rabey¹

2017

JIG

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“…Under the same cultivation condition, the PHA produced from 10 g/L glucose, represented the highest Mw compared to 20 g/L glucose and 10 g/L mixed organic acids. These molecular weights were found to be higher than the PHA polymer synthesized by the recombinant E. coli, expressing a different PHA synthase [44,45]. The highest Mw was achieved from batch fermentation with 10 g/L glucose, whereas the lowest PDI was obtained from the fermentation of mixed acids (Table 6).…”

Section: Polymer Characterizationmentioning

confidence: 91%

Polyhydroxyalkanoate Synthesis by Recombinant Escherichia coli JM109 Expressing PHA Biosynthesis Genes from Comamonas sp. EB172

Yee¹,

Mumtaz²,

Mohammadi³

et al. 2012

J Microb Biochem Technol

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Recombinant Escherichia coli JM109 harbouring the polyhydroxyalkanoate (PHA) biosynthesis gene (phaCABco) of Comamonas sp. EB172, an acid tolerant microbe, was examined for the production of PHAs from various carbon sources. The study demonstrated that the recombinant E. coli JM109 had the potential to utilize both sugar-and acid-based carbon sources, for the biosynthesis of both poly(3-hydroxybutyrate) P(3HB) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) P(3HB-co-3HV) copolymers. In the shake flask experiments, the strain was capable of producing P (3HB-co-3HV) copolymer from mixed organic acids, and higher productivities were obtained using glucose compared to mixed acids. However, PHA accumulation was found to be similar, regardless of the carbon source used. Nitrogen supplementation in the medium was found to improve the cell dry weight, but negatively affected the 3HV formation in copolymer production. Maximum 3HV monomer (3 mol%) was obtained with C/N 42.1, using mixed acids as the carbon source. In the 2L bioreactor, the productivity and yield based on substrate utilization coefficient were found to be 0.16 g PHA/(L.h) and 0.41 g PHA/g substrate under C/N around 75, using 20 g/L glucose and 0.5 g/L ammonium sulphate, respectively. The polymer produced by the recombinant strain had molecular weight in the range of 8.5 x 105 to 1.4 x 106 Da. Overall, the ability of the recombinant E. coli JM109 to utilize both glucose and mixed acids, has widened its substrate selection for fermentation, including the opportunity to use renewable biomass.

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“…in R. rubrum remain unclear. Heterologous expression of class IV PHA synthases has been studied intensively in the past 22,47,48 . In our study, class IV PHA synthases of the PHB accumulating strains Bacillus megaterium and Bacillus cereus were investigated.…”

Section: Phb Accumulation Of Recombinant R Rubrum Strainsmentioning

confidence: 99%

Heterologous Expression of Various PHA Synthase Genes in Rhodospirillum rubrum

Klask

Raberg

Heinrich

et al. 2015

Chem. Biochem. Eng. Q.

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Molecular Weight Change of Polyhydroxyalkanoate (PHA) Caused by the PhaC Subunit of PHA Synthase from Bacillus cereus YB-4 in Recombinant Escherichia coli

Cited by 49 publications

References 26 publications

Isolation, Cloning and Sequencing of Poly 3-Hydroxybutyrate Synthesis Genes from Local Strain of Bacillus cereus mm7 and Expression them in E. coli

Isolation, Cloning and Sequencing of Poly 3-Hydroxybutyrate Synthesis Genes from Local Strain of Bacillus cereus mm7 and Expression them in E. coli

Polyhydroxyalkanoate Synthesis by Recombinant Escherichia coli JM109 Expressing PHA Biosynthesis Genes from Comamonas sp. EB172

Heterologous Expression of Various PHA Synthase Genes in Rhodospirillum rubrum

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