skip to main content
US FlagAn official website of the United States government
dot gov icon
Official websites use .gov
A .gov website belongs to an official government organization in the United States.
https lock icon
Secure .gov websites use HTTPS
A lock ( lock ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.


Title: One-Step Oxidation of Orange Peel Waste to Carbon Feedstock for Bacterial Production of Polyhydroxybutyrate
Orange peels are an abundant food waste stream that can be converted into useful products, such as polyhydroxyalkanoates (PHAs). Limonene, however, is a key barrier to building a successful biopolymer synthesis from orange peels as it inhibits microbial growth. We designed a one-pot oxidation system that releases the sugars from orange peels while eliminating limonene through superoxide (O2• −) generated from potassium superoxide (KO2). The optimum conditions were found to be treatment with 0.05 M KO2 for 1 h, where 55% of the sugars present in orange peels were released and recovered. The orange peel sugars were then used, directly, as a carbon source for polyhydroxybutyrate (PHB) production by engineered Escherichia coli. Cell growth was improved in the presence of the orange peel liquor with 3 w/v% exhibiting 90–100% cell viability. The bacterial production of PHB using orange peel liquor led to 1.7–3.0 g/L cell dry weight and 136–393 mg (8–13 w/w%) ultra-high molecular weight PHB content (Mw of ~1900 kDa) during a 24 to 96 h fermentation period. The comprehensive thermal characterization of the isolated PHBs revealed polymeric properties similar to PHBs resulting from pure glucose or fructose. Our one-pot oxidation process for liberating sugars and eliminating inhibitory compounds is an efficient and easy method to release sugars from orange peels and eliminate limonene, or residual limonene post limonene extraction, and shows great promise for extracting sugars from other complex biomass materials.  more » « less
Award ID(s):
1719875
PAR ID:
10411654
Author(s) / Creator(s):
; ; ;
Date Published:
Journal Name:
Polymers
Volume:
15
Issue:
3
ISSN:
2073-4360
Page Range / eLocation ID:
697
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. ABSTRACT Bacteria and eukaryotes produce the reactive oxygen species superoxide both within and outside the cell. Although superoxide is typically associated with the detrimental and sometimes fatal effects of oxidative stress, it has also been shown to be involved in a range of essential biochemical processes, including cell signaling, growth, differentiation, and defense. Light‐independent extracellular superoxide production has been shown to be widespread among many marine heterotrophs and phytoplankton, but the extent to which this trait is relevant to marine microbial physiology and ecology throughout the global ocean is unknown. Here, we investigate the dark extracellular superoxide production of five groups of organisms that are geographically widespread and represent some of the most abundant organisms in the global ocean. These includeProchlorococcus,Synechococcus,Pelagibacter,Phaeocystis, andGeminigera. Cell‐normalized net extracellular superoxide production rates ranged seven orders of magnitude, from undetectable to 14,830 amol cell−1h−1, with the cyanobacteriumProchlorococcusbeing the lowest producer and the cryptophyteGeminigerabeing the most prolific producer. Extracellular superoxide production exhibited a strong inverse relationship with cell number, pointing to a potential role in cell signaling. We demonstrate that rapid, cell‐number–dependent changes in the net superoxide production rate bySynechococcusandPelagibacterarose primarily from changes in gross production of extracellular superoxide, not decay. These results expand the relevance of dark extracellular superoxide production to key marine microbes of the global ocean, suggesting that superoxide production in marine waters is regulated by a diverse suite of marine organisms in both dark and sunlit waters. 
    more » « less
  2. ABSTRACT There is a growing appreciation within animal and plant physiology that the reactive oxygen species (ROS) superoxide is not only detrimental but also essential for life. Yet, despite widespread production of extracellular superoxide by healthy bacteria and phytoplankton, this molecule remains associated with stress and death. Here, we quantify extracellular superoxide production by seven ecologically diverse bacteria within the Roseobacter clade and specifically target the link between extracellular superoxide and physiology for two species. We reveal for all species a strong inverse relationship between cell-normalized superoxide production rates and cell number. For exponentially growing cells of Ruegeria pomeroyi DSS-3 and Roseobacter sp. strain AzwK-3b, we show that superoxide levels are regulated in response to cell density through rapid modulation of gross production and not decay. Over a life cycle of batch cultures, extracellular superoxide levels are tightly regulated through a balance of both production and decay processes allowing for nearly constant levels of superoxide during active growth and minimal levels upon entering stationary phase. Further, removal of superoxide through the addition of exogenous superoxide dismutase during growth leads to significant growth inhibition. Overall, these results point to tight regulation of extracellular superoxide in representative members of the Roseobacter clade, consistent with a role for superoxide in growth regulation as widely acknowledged in fungal, animal, and plant physiology. IMPORTANCE Formation of reactive oxygen species (ROS) through partial reduction of molecular oxygen is widely associated with stress within microbial and marine systems. Nevertheless, widespread observations of the production of the ROS superoxide by healthy and actively growing marine bacteria and phytoplankton call into question the role of superoxide in the health and physiology of marine microbes. Here, we show that superoxide is produced by several marine bacteria within the widespread and abundant Roseobacter clade. Superoxide levels outside the cell are controlled via a tightly regulated balance of production and decay processes in response to cell density and life stage in batch culture. Removal of extracellular superoxide leads to substantial growth inhibition. These findings point to an essential role for superoxide in the health and growth of this ubiquitous group of microbes, and likely beyond. 
    more » « less
  3. Polyhydroxybutyrate (PHB) is a bio-based, biodegradable alternative to petroleum-based plastics. PHB production at industrial scales remains infeasible, in part due to insufficient yields and high costs. Addressing these challenges requires identifying novel biological chassis for PHB production and modifying known biological chassis to enhance production using sustainable, renewable inputs. Here, we take the former approach and present the first description of PHB production by two prosthecate photosynthetic purple non-sulfur bacteria (PNSB), Rhodomicrobium vannielii and Rhodomicrobium udaipurense. We show that both species produce PHB across photoheterotrophic, photoautotrophic, photoferrotrophic, and photoelectrotrophic growth conditions. Both species show the greatest PHB titers during photoheterotrophic growth on butyrate with dinitrogen gas as a nitrogen source (up to 44.08 mg/L), while photoelectrotrophic growth demonstrated the lowest titers (up to 0.13 mg/L). These titers are both greater (photoheterotrophy) and less (photoelectrotrophy) than those observed previously in a related PNSB, Rhodopseudomonas palustris TIE-1. On the other hand, we observe the highest electron yields during photoautotrophic growth with hydrogen gas or ferrous iron electron donors, and these electron yields were generally greater than those observed previously in TIE-1. These data suggest that non model organisms like Rhodomicrobium should be explored for sustainable PHB production and highlights utility in exploring novel biological chassis. 
    more » « less
  4. This study investigates the underlying mechanisms of hydrogen peroxide (H₂O₂) sensing using a composite material of bismuth oxide and bismuth oxyselenide (Bi2OxSey). The antagonistic effect of tungsten (W)-doping on the electrochemical behavior was also examined. Undoped, 2 mol%, 4 mol%, and 6 mol% W-doped Bi2OxSey nanostructures were synthesized using a one-pot solution phase method involving selenium powder and hydrazine hydrate. W-doping induced a morphological transformation from nanosheets to spherical nanoparticles and amorphization of the bismuth oxyselenide phase. Electrochemical sensing measurements were conducted using cyclic voltammetry (CV) and differential pulse voltammetry (DPV). H₂O₂ detection was achieved over a wide concentration range of 0.02 to 410 µM. In-depth CV analysis revealed the complex interplay of oxidation-reduction processes within the bismuth oxide and Bi2O2Se components of the composite material. W-doping exhibited an antagonistic effect, significantly reducing sensitivity. Among the studied samples, undoped Bi2OxSeγ demonstrated a high sensitivity of 83 μA μM⁻1 cm⁻2 for the CV oxidation peak at 0 V, while 6 mol% W-Bi2OxSey became completely insensitive to H2O2. Interestingly, DPV analysis showed a reversal of sensitivity trends with 2 and 4 mol% W-doping. The applicability of these samples for real-world analysis, including rainwater and urine, was also demonstrated. 
    more » « less
  5. In this study, a facile protocol was used to convert non-valuable orange peels (OP) waste into a new sensing iron oxide orange-peel nanomaterial (FeOP). The presence of iron oxide nanoparticles in the modified OP was confirmed by physicochemical characterisations including Fourier-transform infrared spectroscopy, X-ray diffractometry, thermogravimetry, and scanning electron microscopy-energy dispersive X-ray. FeOP was used to modify a carbon paste electrode (CPE/FeOP) which displayed a significant increase in specific capacitance of 2939 F.g−1, two folds higher than that obtained with CPE at 10 m.s−1 in NaCl. The electroanalysis of Congo red (CR) in aqueous solutions using CPE/FeOP displayed detection limits of 2.8 × 10−7 mol.L−1 and 8.2 × 10−7 mol.L−1 respectively in deionised and spring waters, in the linear range of 5 to 55 µM. CPE/FeOP electrochemical sensor is therefore suitable for the determination of Congo red in wastewater. 
    more » « less