Use In Bioconversion Research Articles

The case for a wider range of flies for use in waste bioconversion

Bioconversion is the process whereby nutrients are recovered from organic waste products, often by flies, to produce value-added products such as protein for animal feed and lipids for biodiesel production. Currently, research and industry focus on a select few fly species for use in bioconversion that are generalists in their feeding behaviour, the black soldier fly,Hermetia illucens L., and the housefly,Musca domestica L. More investigation is needed on species of flies that are more suited to underutilised waste streams, including meat processing waste. Species of flies that breed in carrion, such as blowflies (Calliphoridae) and flesh flies (Sarcophagidae) can be used to reduce meat processing waste and produce a valuable source of protein. In this review, we propose more investigation and use of a wider range of fly species for bioconversion of organic waste. Four blowfly species are recommended for use in the bioconversion of meat processing waste or a mixture of manure waste and meat processing waste.Chrysomya chloropyga (Wiedemann) is a large mammal carcass specialist and has been found to be effective at recovering nutrients from abattoir waste within four days and producing large larvae in the process.Chrysomya putoria (Wiedemann) andChrysomya megacephala (Wiedemann) naturally breed in carrion and faeces, are associated with pit latrines and respond well under mass rearing conditions, with high egg production. These species would be recommended for a large-scale bioconversion facility that receives mixed waste streams including manure and animal remains.Lucilia sericata is known to produce antimicrobial compounds that assist in wound healing and has been frequently bred and studied and responds well to lab and mass rearing conditions. We recognise the potential obstacles to using alternative species in industrial-scale bioconversion facilities and pose future directions for research to overcome these challenges.

Modeling and experimental validation of covalent immobilization of Trametes maxima laccase on glyoxyl and MANA-Sepharose CL 4B supports, for the use in bioconversion of residual colorants.

Our novel strategy for the rational design of immobilized derivatives (RDID) is directed to predict the behavior of the protein immobilized derivative before its synthesis, by the usage of mathematic algorithms and bioinformatics tools. However, this approach needs to be validated for each target enzyme. The objective of this work was to validate the RDID strategy for covalent immobilization of the enzyme laccase from Trametes maxima MUCL 44155 on glyoxyl- and monoaminoethyl-N-aminoethyl (MANA)-Sepharose CL 4B supports. Protein surface clusters, more probable configurations of the protein-supports systems at immobilization pHs, immobilized enzyme activity, and protein load were predicted by RDID1.0 software. Afterward, immobilization was performed and predictions were experimentally confirmed. As a result, the laccase-MANA-Sepharose CL 4B immobilized derivative is better than laccase-glyoxyl-Sepharose CL 4B in predicted immobilized derivative activity (63.6% vs. 29.5%). Activity prediction was confirmed by an experimentally expressed enzymatic activity of 68%, using 2,6-dimethoxyphenol as substrate. Experimental maximum protein load matches the estimated value (11.2±1.3 vs. 12.1protein mg/support mL). The laccase-MANA-Sepharose CL 4B biocatalyst has a high specificity for the acid blue 62 colorant. The results obtained in this work suggest the possibility of using this biocatalyst for wastewater treatment.

Longevity, Fertility and Fecundity of Adult Blow Flies (Diptera: Calliphoridae) Held at Varying Densities: Implications for Use in Bioconversion of Waste.

Large numbers of flies are needed to produce the quantity of larvae required for insect bioconversion of waste. However, this 'mass-rearing' may negatively affect adult survival and reproductive output. This study assessed the suitability for mass-rearing of four blow fly species, Chrysomya chloropyga, Chrysomya chloropyga (Wiedemann), Chrysomya megacephala (F.), Chrysomya putoria (Wiedemann) and Lucilia sericata (Meigen). Flies were kept at densities of 20, 50, 100, 250, 500, and 1,000 flies per 30 × 30 × 30 cm cage with an even sex ratio. Time to 50% mortality (LT50) was recorded, and the effects of density, species, and sex on LT50, fecundity, and fertility were determined. Females survived longer than males across all species. There was evidence for a trade-off between survival and high fecundity in L. sericata and C. chloropyga at density 250. C. megacephala had low fecundity across all densities. At high densities, C. putoria had the lowest mortality and highest fecundity, making it the most suitable for mass-rearing.

Characterization of natural co-cultures of Piromyces with Methanobrevibacter ruminantium from yaks grazing on the Qinghai-Tibetan Plateau: a microbial consortium with high potential in plant biomass degradation

Anaerobic fungi reside in the gut of herbivore and synergize with associated methanogenic archaea to decompose ingested plant biomass. Despite their potential for use in bioconversion industry, only a few natural fungus–methanogen co-cultures have been isolated and characterized. In this study we identified three co-cultures of Piromyces with Methanobrevibacter ruminantium from the rumen of yaks grazing on the Qinghai Tibetan Plateau. The representative co-culture, namely (Piromyces + M. ruminantium) Yak-G18, showed remarkable polysaccharide hydrolase production, especially xylanase. Consequently, it was able to degrade various lignocellulose substrates with a biodegrading capability superior to most previously identified fungus or fungus–methanogen co-culture isolates. End-product profiling analysis validated the beneficial metabolic impact of associated methanogen on fungus as revealed by high-yield production of methane and acetate and sustained growth on lignocellulose. Together, our data demonstrated a great potential of (Piromyces + M. ruminantium) Yak-G18 co-culture for use in industrial bioconversion of lignocellulosic biomass.

Isolation and characterization of bacterial strains with a hydrolytic profile with potential use in bioconversion of agroindustial by-products and waste

There is a trend towards the use of novel technologies nowadays, mainly focused on biological processes, for recycling and the efficient utilization of organic residues that can be metabolized by different microorganisms as a source of energy. In the present study the isolation of bacterial strains from six different agro-industrial by-products and waste was performed with the objective of evaluating their hydrolytic capacities and suitability for use in bioconversion of specific substrates. The 34 isolated strains were screened in specific culture media for the production of various hydrolytic enzymes (lipase, protease, cellulase, and amylase). It was found that 28 strains exhibited proteolytic activity, 18 had lipolytic activity, 13 had caseinolytic activity, 15 had amylolytic activity, and 11 strains exhibited cellulolytic activity. The strains that showed the highest hydrolytic capacities with biotechnological potential were selected, characterized genotipically, and identified as Bacillus, Serratia, Enterococcus, Klebsiella, Stenotrophomonas, Lactococcus, and Escherichia genera. It was concluded that the strain isolates have a high potential for use in the bioconversion of agro-industrial waste, both as a pure culture and as a microbial consortium.

A quantitative method for fungal ligninolytic enzyme screening studies

AbstractScreening is conducted to select a fungus with desired characteristics intended for various applications, e.g. bioremediation and enzyme production. A qualitative method was used in this study for screening of fungal ligninolytic enzyme activities. The fungal ligninolytic activity was correlated with its growth from the screenings to identify a suitable fungus for solid substrate fermentation. Four strains of fungi, namely, Phanerochaete chrysosporium, Pycnoporus sanguineus, Phlebia radiata and Pleurotus sajor‐caju, were screened for their ligninolytic enzyme activities using guaiacol and remazol brilliant blue‐R (RBB‐R) as screening reagents. The screenings were conducted at both, the optimal growth temperature of each fungus and 35 °C. All the fungi, except P. radiata, showed positive guaiacol oxidation and RBB‐R decolourisation activities. On the basis of the results, P. sajor‐caju may be a suitable fungus for use in bioconversion of lignocellulose because it maintained guaiacol oxidation and RBB‐R decolourisation activities at 35 °C. Its halo‐to‐colony area ratios of guaiacol oxidation and RBB‐R decolourisation after 6 days were 7.0 and 2.7, respectively. In the subsequent growth study, the measured colony area was closely correlated with the dry fungal cell weight with a regression value (R2) of 0.971. Thus, the colony size could be used as an indicator of fungal growth if the screening media do not significantly slow the growth of fungus. Copyright © 2010 Curtin University of Technology and John Wiley & Sons, Ltd.

Rapid conversion of toluene by an acinetobacter sp. Tol 5 mutant showing monolayer adsorption to water-oil interface

Two-liquid-phase partitioning bioreactors (TPPBs) consist of an aqueous phase containing microorganism cells and a water-immiscible, non-bioavailable organic phase, and have been investigated for use in bioconversion of toxic, hydrophobic chemicals. We achieved a toluene conversion rate 10-fold faster than any experimental data previously demonstrated, using a novel bacterium in a TPPB employing silicone oil. The bacterium was a bald mutant of toluene-degrading Acinetobacter sp. Tol 5, which has numerous filamentous cell appendages, hydrophobic cell surfaces, adheres to solid surfaces, and self-agglutinates. The bald mutant cells lack the appendages, have decreased adhesiveness to solid surfaces, retain the hydrophobic cell surface, and reversibly adsorb to the surface of oil droplets dispersed in an aqueous phase. The effects of toluene concentration, aqueous and oil phase volume ratios, and cell concentration on the initial conversion rate of toluene were investigated. Volume ratios did not significantly affect conversion rate, and the rate greater than 1 g/(l.h) was attained even in W/O emulsion containing 80% oil. As cell concentration increased, the volumetric conversion rate increased, but the specific conversion rate decreased. The highest volumetric conversion rate, 3.1 g/(l.h), was achieved using 8.20 g/l cells in a TPPB containing 20% oil.

Hexose oxidase from Chondrus crispus: improved purification using perfusion chromatography

1An improved method for purifying hexose oxidase (d-hexose: O2 1-oxidoreductase, EC 1.1.3.5) from the marine red alga Chondrus crispus is described for obtaining enzyme suitable for structural characterization and use in bioconversion of lactose to lactobionic acid. This involved extracting enzyme from finely ground lyophilized tissue in sodium phosphate buffer (pH 7) containing 20% ammonium sulfate, eliminating the previously used solvent extraction and protease treatments, and by applying Poros perfusion chromatography media to achieve rapid separations of high resolution. Primary separation of contaminating phycobiliproteins and carrageenans was achieved using Poros DEAE-50. Sequential HPLC purification steps using Poros HP2 and Poros HQ were followed by Sephacryl S200 h chromatography. Enzyme activity was determined with a peroxidase-coupled assay using 2,2′-azino-bis (3-ethylbenzthiazoline-6 sulfonic acid) substrate. A final specific activity of 69 U/mg was obtained, representing a 100-fold purification with an activity recovery of about 10%. A native size of approximately 117,000 Da was determined by size exclusion chromatography, and SDS-PAGE revealed the presence of 38,000 and 29,000 Da polypeptides that appear to be derived from a 65,000 Da subunit. Further properties of the enzyme are described.

Enzymology and Genetics of Halogenating Enzymes from Bacteria

This paper discusses the properties of bacterial haem-containing and non-haem haloperoxidases, their involvement in the biosynthesis of halometabolites and their use in bioconversion. The very low peroxidase activity of bacterial non-haem haloperoxidases, their stability at high temperature and over a wide pH-range makes them particularly suited for use in the bromination of organic compounds. The chloroperoxidase from Pseudomonas pyrrocinia is the only haloperoxidase showing substrate specificity and regioselectivity. The genes of one chloro- and one bromoperoxidase could be cloned. The corresponding enzymes can now be produced in large amounts and at low costs.