Biotechnology Reports最新文献

The gal80 mutant of yeast Saccharomyces cerevisiae is used for the constitutive expression under strong GAL promoters without galactose induction. To enhance productivity of gal80 mutant, an alternative strain, allgal, was developed by removing all galactose-utilizing genes that consume significant cellular resources in the gal80 strain when cultured in non-galactose conditions. The efficacy of the allgal mutant (gal80, gal1, gal2, gal7, and gal10) was verified by assessing the secretory expression of three recombinant proteins, Candida antarctica lipase B (CalB), human serum albumin (HSA), and human epidermal growth factor (hEGF), using the GAL10 promoter. The growth of the allgal mutant was enhanced by 15–38% compared to the gal80 mutant, and the secretion of recombinant proteins also increased by 16–22% in fed-batch fermentation. Thus, the expression of recombinant proteins using GAL10 promoter in the allgal mutant is suitable for the economical production of recombinant proteins in S. cerevisiae.

Carbendazim (CBZ) is a steady benzimidazole fungicide broadly exploited in cultivation for pre- and post-harvest treatment to dominate microorganisms infection on several plants. CBZ causes toxic effects in the different tissues of rat via influencing biochemical and hematological factors causing histopathological alterations in the liver and kidney of rats. Banana peel (BP) makes up about 38% of the whole banana weight, BP is usually disposed of as waste that is considered as an environmental problem. BP comprise bioactive a lot of compounds that can be exploited for their unique biological and pharmacological attributes. The current study was carried out to determine the protective effect of dried banana peels consumption against carbendazim toxicity in rats. The results indicated that banana peels had the ability to counteract the toxic effect of carbendazim on rats which was evident by the improvement in liver and kidney functions, lipid profile and histopathological examination.

Heat production during fermentation is undesirable. It can affect the growth of biomass, sporulation, products formation and the scaling-up. Physico-chemical properties of substrates influence heat and mass transfer in Solid State Fermentation. Heat is chemically produced into substrates without micro-organism to allow better reproducibility. A 2^4–1 fractional factorial design is chosen to study the influence of four physicochemical parameters on heat transfer: Granulometry, Bulk Density, Carr Index (compressibility index) and Water Absorption Capacity. Results show that the two main physicochemical parameters which influence heat transfer are Granulometry and Carr Index. High Granulometry and low Carr Index have influence on maximum temperature reached during the test, warm-up speed and cooling speed. These two parameters allow efficient air flow through the substrate bed with large interparticle spaces enhancing exchange surface between air and particles. A substrate with these characteristics facilitates heat transfers in forced-aerated reactors.

Coenzyme Q₁₀ (CoQ₁₀) is a powerful antioxidant with a myriad of applications in healthcare and cosmetic industries. The most effective route of CoQ₁₀ production is microbial biosynthesis. In this study, four CoQ₁₀ biosynthesizing purple photosynthetic bacteria: Rhodobacter blasticus, Rhodovulum adriaticum, Afifella pfennigii and Rhodovulum marinum, were identified using 16S rRNA sequencing of enriched microbial mat samples obtained from Purple Island mangroves (Qatar). The membrane bound enzyme 4-hydroxybenzoate octaprenyltransferase (UbiA) is pivotal for bacterial biosynthesis of CoQ₁₀. The identified bacteria could be inducted as efficient industrial bio-synthesizers of CoQ₁₀ by engineering their UbiA enzymes. Therefore, the mutation sites and substitution residues for potential functional enhancement were determined by comparative computational study. Two mutation sites were identified within the two conserved Asp-rich motifs, and the effect of proposed mutations in substrate binding affinity of the UbiA enzymes was assessed using multiple ligand simultaneous docking (MLSD) studies, as a groundwork for experimental studies.

Spontaneous fermentation during black tea production involves several reactions, including the oxidation of phenolic compounds. This process has usually been studied without considering the potential involvement of indigenous tea microorganisms. This work utilised a metagenomic technique targeting bacterial 16S rRNA genes and evaluated the profile of phenolic compounds generated during the production of black tea. The resulting data were used to develop correlational and predictive functional analyses related to bacterial dynamics and the syntheses of various phenolic compounds. In particular, the genera Methylobacterium and Devosia were correlated with gallic acid and quercetin. Concurrently, the genera Sphingomonas, Chryseobacterium and Aureimonas were correlated with kaempferol, theaflavins, thearubigins and theabrownins. These results, supported by predicted functional analysis based on 16S rRNA genes associated with phenolic compounds, indicated that yfiH (polyphenol oxidase) and katG (catalase-peroxidase) are likely the dominant genes of the bacterial community involved in the black tea production process. This research suggests that bacteria could potentially contribute to the production process of black tea.

The purpose of this study was to antibacterial, and antibiofilm activity of two Lactobacillus strains secretome and extraction against E. coli isolated from women with urinary tract infection (UTI). We isolated 100 E. coli samples from women with UTI. Lactobacillus acidophilus and Lactobacillus casei characteristics were evaluated, and their secretome and extraction were prepared. The antibacterial and antibiofilm activity of secretome and extraction of both Lactobacillus strains were evaluated against isolated E. coli samples. L. acidophilus and L. casei were able to tolerate pH 3, bile salts, and pancreatic enzymes. Both probiotics were not resistant to antibiotics and demonstrated an appropriate ability to adhere to the intestinal epithelial cells. Secretome and extraction of L. acidophilus and L. casei strains showed a good antibacterial and antibiofilm against E. coli isolates. Generally, present study suggested that the secretome and extraction of L. acidophilus and L. casei strains exhibits a good antimicrobial activity.

SR4 genotype of rice is high altitude Himalayan rice prone to various abiotic stresses such as cold stress and therefore gives a poor yield. An efficient protocol for callusing and regeneration via direct and indirect means was established using mature seeds as an explant which can be utilized for molecular studies for genetic advancement of Himalayan rice genotype SR4 through transformation. Highest frequency (96.6%) of callus induction was obtained on MS media 3.0 mg/L 2, 4-D. While maximum regeneration frequency (100%), number of shoots with maximum length 9.14 ± 0.204 (cm) from callus was recovered from MS media amended with 5.0 mg/L BAP in combination with 0.5 mg/L NAA with highest number of shoots having an average shoot length 9.14 ± 0.204 (cm) after four weeks of culture. Direct multiple shoot regeneration from seed explants was obtained using various concentrations of TDZ and BAP with highest regeneration frequency was observed on MS media fortified with 6 mg/L of TDZ with maximum number of shoots. The shoots developed roots on MS media supplemented with 0.6 mg/L IBA.

The present article exemplifies a novel method to isolate highly purified bioactive lactoferrin from camel milk. Cytotoxicity of lactoferrin against the Hela cells was used to evaluate its bioactivity. SDS-PAGE and LC-MS analysis was done for its identification and characterization. The purified camel milk lactoferrin was found to be 708 amino acids in length with a molecular weight of 77.3 kDa and a pI value of 8.24. This pH-dependent isolation procedure ensures the retention of bioactive lactoferrin from camel milk. The importance of the present work lies in its simplicity and scalability for manufacturing bioactive lactoferrin at an industrial level.

Fungi are potential biocontrol agents and rich sources of secondary metabolites with demonstrated biological activities. This study aimed to isolate and identify fungi from surface-sterilized honeybees (Apis mellifera), as well as to evaluate their biological activities. One fungal isolate was obtained and identified morphologically and genetically as Mucor bainieri MK-Bee-2. Gas chromatography-mass spectroscopy (GC–MS) analysis of fungus crude extract, showed the existence of six major metabolites representing 92.48% of the total peak area. The crude extract of Mucor bainieri MK-Bee-2 was tested for antimicrobial, antioxidant, and antitumor activities. It demonstrated wide antimicrobial activities against human pathogenic Gram-positive and Gram-negative bacterial strains, as well as Candida albicans, with MIC values ranged from 62.5 to 250 µg/ml. The results revealed that the extract exhibited considerable antioxidant activities indicated by strong inhibition of both DPPH and ABTS free radicals. Additionally, the extract exhibited greater potential anticancer activity against both adenocarcinomic human non-small cell lung cancer cells (A549) [IC₅₀ = 6.45 μg/ml], and immortal cell line hepatoma G2 (HepG2) human liver cancer cells [IC₅₀ = 27.48 μg/ml] and higher selectivity in cancer cells than normal cell lines. Furthermore, the extract showed less cytotoxic activity against normal cells with higher IC₅₀ values of 106.99 and 132.57 μg/ml against human lung fibroblast Wistar-38 (Wi-38) and oral epithelial cells (OEC), respectively. Taken together, the Mucor bainieri MK-Bee-2 extract comprises bioactive compounds as promising potential therapeutic candidates for the treatment of lung cancer. Strikingly, the extract sensitizes the lung cancer cells A549  to the ionizing radiation through the pro-apoptotic pathway as indicated by the annexin V flow cytometry analysis which showed that the extract induced the apoptosis of lung cancer cells.

Three-dimensional cell spheroids are superior cell-administration form for cell-based therapy which generally exhibit superior functionality and long-term survival after transplantation. Here, we nondestructively measured the oxygen consumption rate of cell spheroids using an on-chip electrochemical device (OECD) and examined whether this rate can be used as a marker to estimate the quality of cell spheroids. Cell spheroids containing NanoLuc luciferase-expressing mouse mesenchymal stem cell line C3H10T1/2 (C3H10T1/2/Nluc) were prepared. Spheroids of high or low quality were prepared by altering the medium change frequency. After transplantation into mice, the high-quality C3H10T1/2/Nluc spheroids exhibited a higher survival rate than the low-quality ones. The oxygen consumption rate of the high-quality C3H10T1/2/Nluc spheroids was maintained at high levels, whereas that of the low-quality spheroids decreased with time. These results indicate that OECD-based measurement of the oxygen consumption rate can be used to estimate the quality of cell spheroids without destructive analysis of the spheroids.