Bacterial Production of Poly-β-hydroxybutyrate (PHB): Converting Starch into Bioplastics

Yanti, Nur Arfa; Sembiring, Langkah; Margino, Sebastian; Ahmad, Sitti Wirdhana

doi:10.1007/978-981-16-1823-9_9

Cited by 6 publications

(3 citation statements)

References 42 publications

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“…Ostle & Holt, (1982) report that P(3HB) granules found in bacteria will show orange fluorescence if dropped with a 1% Nile Blue A. This is in accordance with research by Yanti et al, (2021) that bacterial colonies that accumulate PHB in their cells, after Nile blue A staining, show bright orange fluorescence when irradiated with UV light. The discovery of bacteria producing bioplastic P(3HB).…”

Section: Screening Of P(3hb) Bioplastic Producing Bacteria Isolatessupporting

confidence: 79%

Screening of Poly(3-Hydroxybutyrate) P(3HB) Producing Bacteria from Mackerel Fish (Rastrelliger sp.)

Rahmi,

Agustien,

Djamaan

2023

jppipa, pendidikan ipa, fisika, biologi, kimia

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Bioplastic P(3HB) is a plastic that can be decomposed by decomposing microorganisms. Bacteria can produce P(3HB) in conditions of low nutrition and high carbon sources accumulated in cells as energy reserves. Isolation of bacteria-producing bioplastic P(3HB) was carried out in mackerel fish's gills and intestines, allowing direct contact with polluted waters. This research aimed to determine the presence of P(3HB) bioplastic-producing bacteria in mackerel fish samples and to screen for P(3HB) bioplastic-producing bacteria. The stages of the research were the isolation of P(3HB) producing bacteria from the gills and intestines of mackerel fish, screening of P(3HB) bioplastic-producing bacteria by using Nile Blue A staining, and molecular identification of bioplastic-producing bacteria using 16S rRNA gene. The results of bacterial isolation in the gills and intestines of mackerel obtained 10 bacterial isolates. P(3HB) screening found 1 bacterium showing orange fluorescence, namely IKE-1 isolate which was isolated from the gills of mackerel fish and included in the Gram-negative group with a monobacilli cell shape. Molecular identification of bacteria by using the 16S rRNA gene that isolate IKE-1 is Enterobacter roggenkampii. These results indicate that Enterobacter roggenkampii can produce P(3HB) which was confirmed by the screening results

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Section: Screening Of P(3hb) Bioplastic Producing Bacteria Isolatessupporting

confidence: 79%

Screening of Poly(3-Hydroxybutyrate) P(3HB) Producing Bacteria from Mackerel Fish (Rastrelliger sp.)

Rahmi,

Agustien,

Djamaan

2023

jppipa, pendidikan ipa, fisika, biologi, kimia

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“…In most bacteria, PHB production proceeds in three steps mediated by the genes located in the phaCAB operon [ 42 , 43 ]. These genes encode three enzymes including β-ketothiolase (PhaA), acetoacetyl-CoA reductase (PhaB), and PHA synthase (PhaC) [ 44 , 45 ]. However, alternative genes that can function as phaCAB genes were predicted in Salinicola salarius strain ES021 leading to the construction of distinct PHB and MCL-PHA synthetic pathways (Fig.…”

Section: Resultsmentioning

confidence: 99%

Genetic characterization of a novel Salinicola salarius isolate applied for the bioconversion of agro-industrial wastes into polyhydroxybutyrate

Abdelrahman,

Barakat,

Ahmed

2024

Microb Cell Fact

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Background Polyhydroxybutyrate (PHB) has emerged as a promising eco-friendly alternative to traditional petrochemical-based plastics. In the present study, we isolated and characterized a new strain of Salinicola salarius, a halophilic bacterium, from the New Suez Canal in Egypt and characterized exclusively as a potential PHB producer. Further genome analysis of the isolated strain, ES021, was conducted to identify and elucidate the genes involved in PHB production. Results Different PHB-producing marine bacteria were isolated from the New Suez Canal and characterized as PHB producers. Among the 17 bacterial isolates, Salinicola salarius ES021 strain showed the capability to accumulate the highest amount of PHB. Whole genome analysis was implemented to identify the PHB-related genes in Salinicola salarius ES021 strain. Putative genes were identified that can function as phaCAB genes to produce PHB in this strain. These genes include fadA, fabG, and P3W43_16340 (encoding acyl-CoA thioesterase II) for PHB production from glucose. Additionally, phaJ and fadB were identified as key genes involved in PHB production from fatty acids. Optimization of environmental factors such as shaking rate and incubation temperature, resulted in the highest PHB productivity when growing Salinicola salarius ES021 strain at 30°C on a shaker incubator (110 rpm) for 48 h. To maximize PHB production economically, different raw materials i.e., salted whey and sugarcane molasses were examined as cost-effective carbon sources. The PHB productivity increased two-fold (13.34 g/L) when using molasses (5% sucrose) as a fermentation media. This molasses medium was used to upscale PHB production in a 20 L stirred-tank bioreactor yielding a biomass of 25.12 g/L, and PHB of 12.88 g/L. Furthermore, the produced polymer was confirmed as PHB using Fourier-transform infrared spectroscopy (FTIR), gas chromatography-mass spectroscopy (GC–MS), and nuclear magnetic resonance spectroscopy (NMR) analyses. Conclusions Herein,Salinicola salarius ES021 strain was demonstrated as a robust natural producer of PHB from agro-industrial wastes. The detailed genome characterization of the ES021 strain presented in this study identifies potential PHB-related genes. However, further metabolic engineering is warranted to confirm the gene networks required for PHB production in this strain. Overall, this study contributes to the development of sustainable and cost-effective PHB production strategies.

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“…It must be underlined that, potentially, starch could be directly metabolized by Hfx. mediterranei without the need of an intense enzymatic hydrolysis of the bread-derived substrate ( Rodriguez-Valera and Lillo, 1992 ; Huang et al, 2006 ; Yanti et al, 2021 ). Morevover, Hfx.…”

Section: Discussionmentioning

confidence: 99%

Exploitation of wasted bread as substrate for polyhydroxyalkanoates production through the use of Haloferax mediterranei and seawater

et al. 2022

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The use of the halophile microorganism Haloferax mediterranei, able to synthesize poly(hydroxybutyrate-hydroxyvalerate) (PHBV), is considered as a promising tool for the industrial production of bioplastic through bioprocessing. A consistent supplementation of the growth substrate in carbohydrates and minerals is overall necessary to allow its PHBV production. In this work, wasted bread was used as substrate for bioplastic production by microbial fermentation. Instead of the consistent and expensive minerals supplement required for Hfx. mediterranei DSM1411 growth, microfiltered seawater was added to the wasted bread-derived substrate. The suitable ratio of wasted bread homogenate and seawater, corresponding to 40:60, was selected. The addition of proteases and amylase to the bread homogenate promoted the microbial growth but it did not correspond to the increase of bioplastic production by the microorganism, that reach, under the experimental conditions, 1.53 g/L. An extraction procedure of the PHBV from cells, based on repeated washing with water, followed or not by a purification through ethanol precipitation, was applied instead of the conventional extraction with chloroform. Yield of PHBV obtained using the different extraction methods were 21.6 ± 3.6 (standard extraction/purification procedure with CHCl3:H2O mixture), 24.8 ± 3.0 (water-based extraction), and 19.8 ± 3.3 mg PHAs/g of wasted bread (water-based extraction followed by ethanol purification). Slightly higher hydroxyvalerate content (12.95 vs 10.78%, w/w) was found in PHBV obtained through the water-based extraction compared to the conventional one, moreover, the former was characterized by purity of 100% (w/w). Results demonstrated the suitability of wasted bread, supplemented with seawater, to be used as substrate for bioplastic production through fermentation. Results moreover demonstrated that a solvent-free extraction, exclusively based on osmotic shock, could be used to recover the bioplastic from cells.

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Bacterial Production of Poly-β-hydroxybutyrate (PHB): Converting Starch into Bioplastics

Cited by 6 publications

References 42 publications

Screening of Poly(3-Hydroxybutyrate) P(3HB) Producing Bacteria from Mackerel Fish (Rastrelliger sp.)

Screening of Poly(3-Hydroxybutyrate) P(3HB) Producing Bacteria from Mackerel Fish (Rastrelliger sp.)

Genetic characterization of a novel Salinicola salarius isolate applied for the bioconversion of agro-industrial wastes into polyhydroxybutyrate

Exploitation of wasted bread as substrate for polyhydroxyalkanoates production through the use of Haloferax mediterranei and seawater

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