Cloning and heterologous expression of a novel subgroup of class IV polyhydroxyalkanoate synthase genes from the genus <i>Bacillus</i>

Mizuno, Kouhei; Kihara, Takahiro; Tsuge, Takeharu; Lundgren, Benjamin R.; Sarwar, Zaara; Pinto, Atahualpa; Nomura, Christopher T.

doi:10.1080/09168451.2016.1230006

Cited by 10 publications

(7 citation statements)

References 16 publications

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“…4a). The members of the genus Bacillus harbor the type IV PHA synthases (PhaRC) (Mizuno et al 2017). The PhaRC YB4 from Bacillus cereus YB4 demonstrates a boarder substrate specificity than Bacillus megaterium and synthesized PHA copolymers from sugar and plant oils during fermentation (Tsuge et al 2015).…”

Section: Fermentation On the Large Scalementioning

confidence: 99%

Effect of glucose and olive oil as potential carbon sources on production of PHAs copolymer and tercopolymer by Bacillus cereus FA11

et al. 2017

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In this study, the influence of different physicochemical parameters on the yield of polyhydroxyalkanoates (PHAs) produced by Bacillus cereus FA11 is investigated. The physicochemical factors include pH, temperature, time, inoculum size and its age, agitation speed and composition of the glucose rich peptone deficient (GRPD) medium. During two-stage fermentation, B. cereus FA11 produced a significantly high (p < 0.05) yield (80.59% w/w) of PHAs copolymer using GRPD medium containing glucose (15 g/L) and peptone (2 g/L) at pH 7, 30 °C and 150 rpm after 48 h of incubation. On the other hand, the presence of olive oil (1% v/v) and peptone (2 g/L) in the GRPD medium resulted in biosynthesis of tercopolymer during two-stage fermentation and the yield of tercopolymer was 60.31% (w/w). The purified PHAs was characterized by Fourier transform infrared spectroscopy and proton resonance magnetic analysis. Proton resonance magnetic analysis confirmed that the tercopolymer was comprised of three different monomeric subunits, i.e., 3-hydroxybutyrate, 3-hydroxyvalerate and 6-hydroxyhexanoate.

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Section: Fermentation On the Large Scalementioning

confidence: 99%

Effect of glucose and olive oil as potential carbon sources on production of PHAs copolymer and tercopolymer by Bacillus cereus FA11

et al. 2017

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“…The most prevalent PHB biosynthesis process starts with acetyl-CoA and goes through condensation, reduction and polymerization catalyzed by acetyl-CoA acetyltransferase (PhaA), acetoacetyl-CoA reductase (PhaB), PHA synthase (PhaC) respectively for the final PHB production. In another common pathway, enoyl-CoA from fatty acid βoxidation is first oxidized to (R)-3-hydroxyacyl-CoA by (R)-specific enoyl-CoA hydratase (PhaJ) before polymerized into PHB polymer by PhaC [9]. Genes involved in the PHB biosynthesis pathway are co-localized in the chromosome region and organized in an operon.…”

Section: Introductionmentioning

confidence: 99%

Proteome profile changes during poly-hydroxybutyrate intracellular mobilization in gram positive Bacillus cereus tsu1

O’Hair

Thapa

et al. 2020

BMC Microbiol

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Background: Bacillus cereus is a bacterial species which grows efficiently on a wide range of carbon sources and accumulates biopolymer poly-hydroxybutyrate (PHB) up to 80% cell dry weight. PHB is an aliphatic polymer produced and stored intracellularly as a reservoir of carbon and energy, its mobilization is a key biological process for sporulation in Bacillus spp. Previously, B. cereus tsu1 was isolated and cultured on rapeseed cake substrate (RCS), with maximum of PHB accumulation reached within 12 h, and depleted after 48 h. Fore-spore and spore structure were observed after 24 h culture. Results: Quantitative proteomic analysis of B. cereus tsu1 identified 2952 quantifiable proteins, and 244 significantly changed proteins (SCPs) in the 24 h:12 h pair of samples, and 325 SCPs in the 48 h:12 h pair of samples. Based on gene ontology classification analysis, biological processes enriched only in the 24 h:12 h SCPs include purine nucleotide metabolism, protein folding, metal ion homeostasis, response to stress, carboxylic acid catabolism, and cellular amino acid catabolism. The 48 h:12 h SCPs were enriched into processes including carbohydrate metabolism, protein metabolism, oxidative phosphorylation, and formation of translation ternary structure. A key enzyme for PHB metabolism, poly(R)-hydroxyalkanoic acid synthase (PhaC, KGT44865) accumulated significantly higher in 12 h-culture. Sporulation related proteins SigF and SpoEII were significantly higher in 24 h-samples. Enzymes for nitrate respiration and fermentation accumulated to the highest abundance level in 48 h-culture. Conclusions: Changes in proteome of B. cereus tsu1 during PHB intracellular mobilization were characterized in this study. The key enzyme PhaC for PHB synthesis increased significantly after 12 h-culture which supports the highest PHB accumulation at this time point. The protein abundance level of SpoIIE and SigF also increased, correlating with sporulation in 24 h-culture. Enzymes for nitrate respiration and fermentation were significantly induced in 48 hculture which indicates the depletion of oxygen at this stage and carbon flow towards fermentative growth. Results from this study provide insights into proteome profile changes during PHB accumulation and reuse, which can be applied to achieve a higher PHB yield and to improve bacterial growth performance and stress resistance.

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“…The most prevalent PHB biosynthesis pathway starts with acetyl-CoA and goes through condensation, reduction and polymerization catalyzed by acetyl-CoA acetyltransferase (PhaA), acetoacetyl-CoA reductase (PhaB), PHA synthase (PhaC) respectively for the final PHB production. In another common pathway, enoyl-CoA from fatty acid β-oxidation is first oxidized to (R)-3-hydroxyacyl-CoA by (R)-specific enoyl-CoA hydratase (PhaJ) before polymerized into PHB polymer by PhaC [9]. The genes encoding for PHB biosynthesis enzymes are often co-localized and organized in an operon.…”

Section: Introductionmentioning

confidence: 99%

Proteome Profile Changes During Poly-hydroxybutyrate Intracellular Mobilization in Gram Positive Bacillus cereus tsu1

O’Hair

Thapa

et al. 2020

Preprint

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Background Bacillus cereus is a bacterial species which grows efficiently on a wide range of carbon sources and accumulates biopolymer poly-hydroxybutyrate (PHB) up to 80% cell dry weight. PHB is an aliphatic polymer produced and stored intracellularly as a reservoir of carbon and energy, its mobilization is a key biological process for sporulation in Bacillus spp. Previously, a B. cereus tsu1 was isolated and cultured on rapeseed cake substrate (RCS), with maximum of PHB accumulation reached within 12 h, and depleted after 48 h; fore-spore and spore structure were observed after 24 h culture. Results Quantitative proteomic analysis of B. cereus tsu1 identified 2,952 quantifiable proteins, and 244 significantly changed proteins (SCPs) in the 24h-12h pair of samples, 325 SCPs in the 48h-12h pair of samples. Based on gene ontology classification analysis, biological processes enriched only in the 24h:12h SCPs include purine nucleotide metabolism, protein folding, metal ion homeostasis, response to stress, carboxylic acid catabolism, and cellular amino acid catabolism. The 48h:12h SCPs were enriched into processes including carbohydrate metabolism, protein metabolism, oxidative phosphorylation, and formation of translation ternary structure. A key enzyme for PHB metabolism, poly(R)-hydroxyalkanoic acid synthase (PhaC, KGT44865) accumulated significantly higher in 12h-culture. Sporulation related proteins SigF and SpoEII were significantly higher in 24h-samples. Enzymes for nitrate respiration and fermentation accumulated at a higher level in 48h-culture. Conclusions Changes in proteome of B. cereus tsu1 during PHB intracellular mobilization were characterized in this study. The key enzyme PhaC for PHB synthesis increased significantly after 12h-culture which supports the highest PHB accumulation at this time point. The protein abundance level of SpoIIE and SigF were increased which is correlated with sporulation in 24h-culture. Enzymes for nitrate respiration and fermentation were significantly induced in 48h-culture which indicates the depletion of oxygen at this stage and carbon flow towards fermentative growth. Results from this study provide insights into proteome profile changes during PHB accumulation and reuse, which can be applied to achieve a higher PHB yield and to improve bacterial growth performance and stress resistance.

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Cloning and heterologous expression of a novel subgroup of class IV polyhydroxyalkanoate synthase genes from the genus Bacillus

Cited by 10 publications

References 16 publications

Effect of glucose and olive oil as potential carbon sources on production of PHAs copolymer and tercopolymer by Bacillus cereus FA11

Effect of glucose and olive oil as potential carbon sources on production of PHAs copolymer and tercopolymer by Bacillus cereus FA11

Proteome profile changes during poly-hydroxybutyrate intracellular mobilization in gram positive Bacillus cereus tsu1

Proteome Profile Changes During Poly-hydroxybutyrate Intracellular Mobilization in Gram Positive Bacillus cereus tsu1

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