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Improving PHA production in a SBR of coupling PHA-storing microorganism enrichment and PHA accumulation by feed-on-demand control.

ABSTRACT: With volatile fatty acids as substrates, the typical polyhydroxyalkanoates (PHAs) production by mixed culture always includes two steps: PHA-storing culture enrichment via aerobic dynamic feeding strategy and PHA accumulation under nutrient-limited condition. To simplify the PHA-production steps, the enrichment and accumulation step were coupled in a SBR. At start-up period, to investigate the effect of settling selection, one acetate-fed SBR was operated by settling selection-double growth limitation (SS-DGL) strategy, while the other was operated by DGL strategy. The results showed that the stable operation in SBR1 was obtained at about 21, 12 days faster than SBR2, implying the settling selection accelerated the start-up process. After omitting the settling selection under the stable operation, the SBR1 was run above 15 days. The results showed that the performance was not substantial altered. Therefore, the settling selection affected the start-up process but not the stable operation. At operational period, based on the sharp decreasing of oxygen uptake rate (OUR), the poly-?-hydroxybutyrate (PHB) content was improved 13%, from 70 to 83% by feed-on-demand control-double growth limitation (FD-DGL). And the harvested volumetric productivity was 5.0 gPHB/L/day, almost 1-folder improvement. That was to say, the PHB production in a SBR of coupling the enrichment and accumulation step was improved by feed-on-demand control. Meanwhile, the FD experiment can keep steady running for 10 SRTs. Therefore, the SS-DGL/FD-DGL strategy was a promising method for PHA production.

SUBMITTER: Zeng S

PROVIDER: S-EPMC5997608 | biostudies-literature | 2018 Jun

REPOSITORIES: biostudies-literature

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Improving PHA production in a SBR of coupling PHA-storing microorganism enrichment and PHA accumulation by feed-on-demand control.

Zeng Shanwen S Song Fuzhong F Lu Peili P He Qiang Q Zhang Daijun D

AMB Express 20180612 1

With volatile fatty acids as substrates, the typical polyhydroxyalkanoates (PHAs) production by mixed culture always includes two steps: PHA-storing culture enrichment via aerobic dynamic feeding strategy and PHA accumulation under nutrient-limited condition. To simplify the PHA-production steps, the enrichment and accumulation step were coupled in a SBR. At start-up period, to investigate the effect of settling selection, one acetate-fed SBR was operated by settling selection-double growth limi ...[more]

PMID: 29896682

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Project description:Seven strains of sulfate-reducing bacteria (SRB) were tested for the accumulation of polyhydroxyalkanoates (PHAs). During growth with benzoate Desulfonema magnum accumulated large amounts of poly(3-hydroxybutyrate) [poly(3HB)]. Desulfosarcina variabilis (during growth with benzoate), Desulfobotulus sapovorans (during growth with caproate), and Desulfobacterium autotrophicum (during growth with caproate) accumulated poly(3HB) that accounted for 20 to 43% of cell dry matter. Desulfobotulus sapovorans and Desulfobacterium autotrophicum also synthesized copolyesters consisting of 3-hydroxybutyrate and 3-hydroxyvalerate when valerate was used as the growth substrate. Desulfovibrio vulgaris and Desulfotalea psychrophila were the only SRB tested in which PHAs were not detected. When total DNA isolated from Desulfococcus multivorans and specific primers deduced from highly conserved regions of known PHA synthases (PhaC) were used, a PCR product homologous to the central region of class III PHA synthases was obtained. The complete pha locus of Desulfococcus multivorans was subsequently obtained by inverse PCR, and it contained adjacent phaE(Dm) and phaC(Dm) genes. PhaC(Dm) and PhaE(Dm) were composed of 371 and 306 amino acid residues and showed up to 49 or 23% amino acid identity to the corresponding subunits of other class III PHA synthases. Constructs of phaC(Dm) alone (pBBRMCS-2::phaC(Dm)) and of phaE(Dm)C(Dm) (pBBRMCS-2::phaE(Dm)C(Dm)) in various vectors were obtained and transferred to several strains of Escherichia coli, as well as to the PHA-negative mutants PHB(-)4 and GPp104 of Ralstonia eutropha and Pseudomonas putida, respectively. In cells of the recombinant strains harboring phaE(Dm)C(Dm) small but significant amounts (up to 1.7% of cell dry matter) of poly(3HB) and of PHA synthase activity (up to 1.5 U/mg protein) were detected. This indicated that the cloned genes encode functionally active proteins. Hybrid synthases consisting of PhaC(Dm) and PhaE of Thiococcus pfennigii or Synechocystis sp. strain PCC 6308 were also constructed and were shown to be functionally active.

Project description:Pseudomonas chlororaphis PA23 was isolated from the rhizosphere of soybeans and identified as a biocontrol bacterium against Sclerotinia sclerotiorum, a fungal plant pathogen. This bacterium produces a number of secondary metabolites, including phenazine-1-carboxylic acid, 2-hydroxyphenazine, pyrrolnitrin (PRN), hydrogen cyanide, proteases, lipases and siderophores. It also synthesizes and accumulates polyhydroxyalkanoate (PHA) polymers as carbon and energy storage compounds under nutrient-limited conditions. Pseudomonads like P. chlororaphis metabolize glucose via the Entner-Doudoroff and Pentose Phosphate pathways, which provide precursors for phenazine production. Mutants defective in phenazine (PHZ; PA23-63), PRN (PA23-8), or both (PA23-63-1) accumulated higher concentrations of PHAs than the wild-type strain (PA23) when cultured in Ramsay's Minimal Medium with glucose or octanoic acid as the carbon source. Expression levels of six pha genes, phaC1, phaZ, phaC2, phaD, phaF, and phaI, were compared with wild type PA23 by quantitative real time polymerase chain reaction (qPCR). The qPCR studies indicated that there was no change in levels of transcription of the PHA synthase genes phaC1 and phaC2 in the phz? (PA23-63) and phz? prn? (PA23-63-1) mutants in glucose medium. There was a significant increase in expression of phaC2 in octanoate medium. Transcription of phaD, phaF and phaI increased significantly in the phz? prn? (PA23-63-1) mutant. Mutations in regulatory genes like gacS, rpoS, and relA/spoT, which affect PHZ and PRN production, also resulted in altered gene expression. The expression of phaC1, phaC2, phaF, and phaI genes was down-regulated significantly in gacS and rpoS mutants. Thus, it appears that PHZ, PRN, and PHA production is regulated by common mechanisms. Higher PHA production in the phz? (PA23-63), prn- (PA23-8), and phz? prn? (PA23-63-1) mutants in octanoic medium could be correlated with higher expression of phaC2. Further, the greater PHA production observed in the phz? and prn? mutants was not due to increased transcription of PHA synthase genes in glucose medium, but due to more accessibility of carbon substrates and reducing power, which were otherwise used for the synthesis of PHZ and PRN.

Dataset Information

Improving PHA production in a SBR of coupling PHA-storing microorganism enrichment and PHA accumulation by feed-on-demand control.

Publications

Improving PHA production in a SBR of coupling PHA-storing microorganism enrichment and PHA accumulation by feed-on-demand control.

Similar Datasets

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