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Valorization Potential of a Novel Bacterial Strain, Bacillus altitudinis RSP75, towards Lignocellulose Bioconversion: An Assessment of Symbiotic Bacteria from the Stored Grain Pest, Tribolium castaneum.

ABSTRACT: Bioconversion of lignocellulose into renewable energy and commodity products faces a major obstacle of inefficient saccharification due to its recalcitrant structure. In nature, lignocellulose is efficiently degraded by some insects, including termites and beetles, potentially due to the contribution from symbiotic gut bacteria. To this end, the presented investigation reports the isolation and characterization of cellulolytic bacteria from the gut system of red flour beetle, Tribolium castaneum. Out of the 15 isolated bacteria, strain RSP75 showed the highest cellulolytic activities by forming a clearance zone of 28 mm in diameter with a hydrolytic capacity of ~4.7. The MALDI-TOF biotyping and 16S rRNA gene sequencing revealed that the strain RSP75 belongs to Bacillus altitudinis. Among the tested enzymes, B. altitudinis RSP75 showed maximum activity of 63.2 IU/mL extract for xylanase followed by β-glucosidase (47.1 ± 3 IU/mL extract) which were manifold higher than previously reported activities. The highest substrate degradation was achieved with wheat husk and corn cob powder which accounted for 69.2% and 54.5%, respectively. The scanning electron microscopy showed adhesion of the bacterial cells with the substrate which was further substantiated by FTIR analysis that depicted the absence of the characteristic cellulose bands at wave numbers 1247, 1375, and 1735 cm^-1 due to hydrolysis by the bacterium. Furthermore, B. altitudinis RSP75 showed co-culturing competence with Saccharomyces cerevisiae for bioethanol production from lignocellulose as revealed by GC-MS analysis. The overall observations signify the gut of T. castaneum as a unique and impressive reservoir to prospect for lignocellulose-degrading bacteria that can have many biotechnological applications, including biofuels and biorefinery.

SUBMITTER: Dar MA

PROVIDER: S-EPMC8468446 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:In spite of great developments in the agricultural field and plant productivity in the last decades, the concern about the control of agricultural pests is still continuous. However, pest management is expected to have more effects from nanomaterials by providing innovative solutions. The current study confirms the biotransformation of copper nanoparticles (CuNPs) using a cell-free culture extract of metal copper-resistant bacteria Pseudomonas fluorescens MAL2, which was isolated from heavy metal-contaminated soils collected from Sharqia Governorate, Egypt. The local screened bacterial isolate, Pseudomonas fluorescens MAL2, is similar to Pseudomonas fluorescens DSM 12442T DSM. After optimization of growth conditions, F-Base medium was found to be the best medium and pH 7, temperature 35 °C, concentration of CuSO4·5H2O 300 ppm, 10 mL supernatant: 40 mL CuSO4·5H2O (300 ppm), and reaction time 90 min were recorded as the best growth conditions to the fabrication of CuNPs. The formed CuNPs were characterized using initially visual observation of the change in the color of the reaction mixture from blue color to the dark green as an indication of CuNPs biotransformation. Then, UV-Vis spectroscopy showed a maximum absorption at 610 nm under the optimum conditions performed. Transmission Electron Microscopy (TEM) revealed the formation of spherical aspect with size ranges from 10:70 nm; moreover, Energy Dispersive X-ray spectroscopy (EDX) indicated the presence of CuNPs and other elements. In addition, the presence of alcohols, phenols, alkenes, and amines is confirmed by Fourier-Transform Infrared spectroscopy (FTIR) spectroscopy analysis. Dynamic Light Scattering (DLS) supported that the Zeta-average size of nanoparticle was 48.07 with 0.227 PdI value. The Zeta potential showed -26.00mV with a single peak. The biosynthesized CuNPs (Bio CuNPs) showed toxicity against the stored grain pest (Tribolium castaneum), where LC50 value was 37 ppm after 5 days of treatment. However, the negligible effect was observed with chemical synthesis of CuNPs (Ch CuNPs) at the same concentration. The results suggest that Bio CuNPs could be used not only as a biocontrol agent, but also as an ecofriendly and inexpensive approach for controlling the stored grain pests.

Project description:Research backgroundThe presence of insect fragments is one of the major constrains in stored food commodities and it causes considerable loss in the quality of the produce. The management of the pest is viewed as a huge challenge in foodprocessingindustry. Conventionally, the detection of Tribolium castenaum in the food processing industry is carried out by acid hydrolysis and staining methods that are time consuming and lack precision.Experimental approachConsidering the importance of a quick and effective method, a quantitative polymerase chain reaction (qPCR)-based approach was developed and elucidated in this study. The mitochondrial cytochrome oxidase I (mtCOI) gene was identified as a target due to its abundance in the pest. Specific primers were designed against the target gene by Primer Premier software and amplified in a qPCR.Results and conclusionsThis method is capable of detecting all the ontogenic stages of T. castaneum in stored wheat flour. Earlier experiments had demonstrated that about 20 µg of DNA can be obtained from 2.2 mg of insects. To quantify the infestation levels, the cycle threshold (Ct) values obtained from known samples were subjected to regression analysis and expressed as adult equivalents. In the unknown samples, the infestation was calculated as 1.74 and 0.046 adult insects in 5 g of wheat flour. The maximum permissible limit of insect fragments in flour is 75 insect fragments or approx. 3 adults per 50 g of flour as per the US Food and Drug Administration (FDA). Hence, by adopting this new method, it is possible for the warehouse operators to arrive at a decision to proceed with efficient management practices where wheat flour is stored. Also, this method can be ratified by government agencies associated with international business to ascertain whether the wheat flour meets the standards set by the respective country before subjecting to foreign trade.Novelty and scientific contributionThis study is the first of its kind in the detection and quantification of T. castaneum in milled products. So far, only conventional methods have been employed to assess the presence of the pests and manual counting of fragments are practiced to quantify the infestation levels. The developed qPCR method is faster, reliable and can be employed in milling industries, bakery industries, food processing plants and foreign trade units for critical detection and quantification of T. castaneum pest infestation.

Dataset Information

Valorization Potential of a Novel Bacterial Strain, Bacillus altitudinis RSP75, towards Lignocellulose Bioconversion: An Assessment of Symbiotic Bacteria from the Stored Grain Pest, Tribolium castaneum.

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