Abstract The increasing interest in environmental protection laws has compelled companies to regulate the disposal of waste organic materials. Despite efforts to explore alternative energy sources, the world remains heavily dependent on crude petroleum oil and its derivatives. The expansion of the petroleum industry has significant implications for human and environmental well-being. Bioremediation, employing living microorganisms, presents a promising approach to mitigate the harmful effects of organic hydrocarbons derived from petroleum. This study aimed to isolate and purify local yeast strains from oil-contaminated marine water samples capable of aerobically degrading crude petroleum oils and utilizing them as sole carbon and energy sources. One yeast strain (isolate B) identified as Candida tropicalis demonstrated high potential for biodegrading petroleum oil in seawater. Physiological characterization revealed the strain’s ability to thrive across a wide pH range (4–11) with optimal growth at pH 4, as well as tolerate salt concentrations ranging from 1 to 12%. The presence of glucose and yeast extract in the growth medium significantly enhanced the strain's biomass formation and biodegradation capacity. Scanning electron microscopy indicated that the yeast cell diameter varied based on the medium composition, further emphasizing the importance of organic nitrogenous sources for initial growth. Furthermore, the yeast strain exhibited remarkable capabilities in degrading various aliphatic and aromatic hydrocarbons, with a notable preference for naphthalene and phenol at 500 and 1000 mg/l, naphthalene removal reached 97.4% and 98.6%, and phenol removal reached 79.48% and 52.79%, respectively. Optimization experiments using multi-factorial sequential designs highlighted the influential role of oil concentration on the bioremediation efficiency of Candida tropicalis strain B. Moreover, immobilized yeast cells on thin wood chips demonstrated enhanced crude oil degradation compared to thick wood chips, likely due to increased surface area for cell attachment. These findings contribute to our understanding of the potential of Candida tropicalis for petroleum oil bioremediation in marine environments, paving the way for sustainable approaches to address oil pollution.
A shaken flask cultivation condition for enhanced and accelerated beta-mannanase formation by Bacillus subtilis 168 was achieved. Among five examined fermentation media a formula that supported enzyme generation and retarded biomass yield and sporulation was selected. The deficiency of biomass production in this medium was mastered by choosing a seed culture medium that accelerated growth and initiation of beta-mannanase synthesis. With respect to enzyme production, the optimum pH and temperature were 7 and 37 degrees C, respectively. The biosynthesis of the enzyme was extremely influenced by the cell growth state as a modulator, glucose as a catabolite repressor, and galactomannan as an inducer. A galactomannan concentration of 4 g l-1 induced a beta-mannanase activity level of 17.5 U ml-1 after 24 h of incubation at the experimental condition. Higher inducer concentrations supported growth rather than enzyme production. The influence of inoculum size was so remarkable that, at optimum, a crude filtrate with an enzyme activity of 33U ml-1 was yielded within 4 hours. It appears that this is among the highest rates reported for beta-mannanase production. We have also demonstrated that blocking of the sporulation process at stage II do not affect enzyme production significantly. This would allow an extended enzyme production phase especially in a continuous culture.
A Bacillus subtilis wild type strain and a kinA (spoIIJ) isogenic mutant were compared as hosts for the expression of the Escherichia coli beta-galactosidase gene, lacZ, driven by the B. subtilis aprE promoter in a chromosomal system. The 2 x SG sporulation formula, with some modifications, was used as a basal medium. The specific activity values recorded by the mutant strain at the stationary phase were markedly higher than those achieved by the wild type host. Exposure of the cells to increasing levels of chloramphenicol resulted in significant amplifications of the lacZ region. Gene copy numbers of 19 and 11 were estimated in the amplified wild type and kinA strains, respectively, with high segregational stability records. The magnitude of beta-galactosidase over-expression was dependent on, and roughly proportional to antibiotic resistance levels. Among five examined by-products, a 2.3-times diluted concentration of neutralized cheese whey was successfully used as a sole medium component for over-expression of the recombinant beta-galactosidase gene in B. subtilis.
Herein, we introduce an innovative nanohybrid material for advanced wastewater treatment, composed of Corchorus olitorius-derived biochar and bismuth oxychloride (Biochar/Bi12O17Cl2), demonstrated in a solar photoreactor. This work focuses on the efficient degradation of linezolid (LIN), a persistent pharmaceutical pollutant, utilizing the unique (photo)catalytic capabilities of the nanohybrid. Compared with its individual components, the biochar/Bi12O17Cl2 hybrid exhibits a remarkable degradation efficiency of 82.6% for LIN, alongside significant chemical oxygen demand (COD) and total organic carbon (TOC) mineralization rates of 81.3 and 75.8%, respectively. These results were achieved within 3 h under solar irradiation, using an optimal composite dose of 125 mg/L at pH 4.3 ± 0.45, with an initial COD and LIN concentrations of 1605 and 160.8 mg/L and TOC of 594.3 mg/L. The nanohybrid's stability across five cycles of use demonstrates its potential for repeated applications, with degradation efficiencies of 82.6 and 77.9% in the first and fifth cycles, respectively. This indicates the biochar/Bi12O17Cl2 composite's suitability as a sustainable and cost-effective solution for the remediation of heavily contaminated waters. Further, the degradation pathway proposed the degradation of all of the generated intermediates to a single-ring compound. Contributing to the development of next-generation materials for environmental remediation, this research underscores the critical role of nanotechnology in enhancing water quality and ecosystem sustainability and addressing the global imperative for clean water access and environmental preservation.
A Bacillus subtilis wild type strain and a kinA (spoIIJ) isogenic mutant were compared as hosts for the expression of the Escherichia coli beta-galactosidase gene, lacZ, driven by the B. subtilis aprE promoter in a chromosomal system. The 2 x SG sporulation formula, with some modifications, was used as a basal medium. The specific activity values recorded by the mutant strain at the stationary phase were markedly higher than those achieved by the wild type host. Exposure of the cells to increasing levels of chloramphenicol resulted in significant amplifications of the lacZ region. Gene copy numbers of 19 and 11 were estimated in the amplified wild type and kinA strains, respectively, with high segregational stability records. The magnitude of beta-galactosidase over-expression was dependent on, and roughly proportional to antibiotic resistance levels. Among five examined by-products, a 2.3-times diluted concentration of neutralized cheese whey was successfully used as a sole medium component for over-expression of the recombinant beta-galactosidase gene in B. subtilis.
Bivalve aquaculture is generally influenced by bacterial pathogens that cause high mortality-related losses in hatcheries. Six bacterial phenotypes had been previously isolated as the most predominant microorganisms in carpet shell clam ( Tapes decussates ) samples collected from natural beds in representative Egyptian clam fisheries. The main aims of this study were to investigate the nature of the pathogenic strains that significantly affect clam survival and test their susceptibility to commercially available antibiotics. Based on their 16S rRNA sequences and some biochemical features, two potent clam pathogens were emerged; one of which is gram-positive and the other is gram-negative. The former isolate was identified as Micrococcus luteus and the latter as Vibrio alginolyticus . Experimental challenges with the two bacterial pathogens introduced at different initial cell concentrations (2.5 X 10 4 - 7.5 X 10 4 cfu ml -1 ) showed markedly diverse clam mortality results. However, the pathogenic interaction of M. luteus with clam survival was generally higher than that of V. alginolyticus . When introduced at a relatively low initial cell density, the infectious records of V. alginolyticus were significantly increased suggesting that the expression of its key virulence factors is mainly triggered as a response to host contact. Antibiotic susceptibility tests suggested chloramphenicol and tetracycline as markedly effective agents that can be used to control the spread of these two bacterial pathogens in aquaculture applications.
Cell-free (in vitro) expression is a robust alternative platform to the cell-based (in vivo) system for recombinant protein production. Tumor necrosis factor-alpha (TNF-α) is an effective pro-inflammatory cytokine with pleiotropic effects. The aim of the current study was de novo optimized expression of soluble and active human TNF-α by an in vitro method in an E. coli-based cell-free protein synthesis (CFPS) system and its biological activity evaluation. The codon-optimized synthetic human TNF-α gene was constructed by a two-step PCR, cloned into pET101/D-TOPO vector and then expressed by the E. coli CFPS system. Cell-free expression of the soluble protein was optimized using a response surface methodology (RSM). The anticancer activity of purified human TNF-α was assessed against three human cancer cell lines: Caco-2, HepG-2 and MCF-7. Data from RSM revealed that the lowest value (7.2 µg/mL) of cell-free production of recombinant human TNF-α (rhTNF-α) was obtained at a certain incubation time (6 h) and incubation temperature (20 °C), while the highest value (350 µg/mL) was recorded at 4 h and 35 °C. This rhTNF-α showed a significant anticancer potency. Our findings suggest a cell-free expression system as an alternative platform for producing soluble and functionally active recombinant TNF-α for further research and clinical trials.
Abstract Herein, a novel composite of Corchorus olitorius-derived biochar and Bi 12 O 17 Cl 2 was fabricated and utilized for the degradation of tetracycline (TC) in a solar photo-oxidation reactor. The morphology, chemical composition, and interaction between the composite components were studied using various analyses. The biochar showed a TC removal of 52.7% and COD mineralization of 59.6% using 150 mg/L of the biochar at a pH of 4.7 ± 0.5, initial TC concentration of 163 mg/L, and initial COD of 1244 mg/L. The degradation efficiency of TC increased to 63% and the mineralization ratio to 64.7% using 150 mg/L of bare Bi 12 O 17 Cl 2 at a pH of 4.7 ± 0.5, initial TC concentration of 178 mg/L, and COD of 1034 mg/L. In the case of biochar/Bi 12 O 17 Cl 2 composite, the degradation efficiency of TC and COD mineralization ratio improved to 85.8% and 77.7% due to the potential of biochar to accept electrons which retarded the recombination of electrons and holes. The synthesized composite exhibited high stability over four succeeding cycles. According to the generated intermediates, TC could be degraded to caprylic acid and pentanedioic acid via the frequent attack by the reactive species. The prepared composite is a promising photocatalyst and can be applied in large-scale systems due to its high degradation and mineralization performance in a short time besides its low cost and stability.
A Bacillus subtilis wild type strain and a kinA (spoIIJ) isogenic mutant were compared as hosts for the expression of the Escherichia coli β-galactosidase gene, lacZ, driven by the B. subtilis aprE promoter in a chromosomal system. The 2 × SG sporulation formula, with some modifications, was used as a basal medium. The specific activity values recorded by the mutant strain at the stationary phase were markedly higher than those achieved by the wild type host. Exposure of the cells to increasing levels of chloramphenicol resulted in significant amplifications of the lacZ region. Gene copy numbers of 19 and 11 were estimated in the amplified wild type and kinA strains, respectively, with high segregational stability records. The magnitude of β-galactosidase over-expression was dependent on, and roughly proportional to antibiotic resistance levels. Among five examined by-products, a 2.3-times diluted concentration of neutralized cheese whey was successfully used as a sole medium component for over-expression of the recombinant β-galactosidase gene in B. subtilis.
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