Aspergillus spp., a versatile cell factory for enzymes and metabolites: Interventions through genome editing

Aspergillus sp. is widely distributed in nature and plays significant roles in the degradation of lignocellulose biomass and extensively used in bioprocess and fermentation technology and many species are also a generally regarded safe. Many of the Aspergillus species are established cell factories due to their inherent capacity in secreting large number of hydrolytic enzymes. With the advent of next generation genomic technologies and metabolic engineering technologies, the production potential of Aspergillus cell factory has improved over the years. Various genome editing tools has been developed for Aspergillus like engineered nucleases, zinc finger nucleases, TALEN and CRISPR-Cas9 system. Currently, the CRISPR/Cas9-based technique is extensively used to enhance the effectiveness of gene manipulation in model system Aspergillus nidulans and other strains like Aspergillus oryzae, Aspergillus niger and Aspergillus fumigatus. This review describes the recent developments of genome editing technologies in Aspergillus the synthesis of heterologous proteins and secondary metabolites in the Aspergillus species

Bioconversion of waste cooking oil for the production of poly-3-hydroxybutyrate using Bacillus cereus MPTDC

557-562Used cooking oil is generated as a byproduct during frying process. It cannot be reused for cooking process due to health issues such as cancer and other digestive disorders. Alternative strategy is utilization of this waste cooking oil for production of poly-3-hydroxybutyrate (PHB) a biopolymer which can be used as a substitute for petroleum derived plastics or other value added products. In the present investigation, we used waste cooking oil as carbon source for PHB production by Bacillus cereus MPTDC. The optimum conditions of PHB production by Bacillus cereus MPTDC were waste cooking oil concentration of 2% (v/v), incubation time of 96 h, ammonium sulphate concentration of 7.5% and yeast extract concentration of 0.2%. Under optimized conditions the strain produced 3.777 g/L of PHB. The results indicate the potential of used cooking oil as carbon source for PHB production by Bacillus cereus MPTDC

Bioconversion of waste cooking oil for the production of poly-3-hydroxybutyrate using Bacillus cereus MPTDC

Used cooking oil is generated as a byproduct during frying process. It cannot be reused for cooking process due to health issues such as cancer and other digestive disorders. Alternative strategy is utilization of this waste cooking oil for production of poly-3-hydroxybutyrate (PHB) a biopolymer which can be used as a substitute for petroleum derived plastics [ABG1] or other value added products. In the present investigation, we used waste cooking oil as carbon source for PHB production by Bacillus cereus MPTDC. The optimum conditions of PHB production by Bacillus cereus MPTDC were waste cooking oil concentration of 2% (v/v), incubation time of 96 h, ammonium sulphate concentration of 7.5% and yeast extract concentration of 0.2%. Under optimized conditions the strain produced 3.777 g/L of PHB. The results indicate the potential of used cooking oil as carbon source for PHB production by Bacillus cereus MPTDC

Applications of Microbial Enzymes in Food Industry

Uporaba enzima i mikroorganizama za pripremu hrane poznata je od davnina. S napretkom tehnologije razvijeni su novi enzimi specifičnih svojstava i širokog raspona primjene, te se neprestano traga za novim mogućnostima njihove uporabe. Bakterije, kvasci i gljivice te njihovi enzimi često se upotrebljavaju za pripremu hrane poboljšanog okusa i teksture, a ekonomski su isplativi. Mikrobni enzimi se koriste u većoj mjeri nego biljni i životinjski enzimi, i to zbog jednostavnije i jeftinije proizvodnje te njihove postojane kvalitete. U ovom se revijalnom prikazu raspravlja o najnovijim postignućima u tehnologiji proizvodnje enzima u prehrambenoj industriji. Naveden je opsežan popis enzima koji se koriste za obradu hrane, mikroorganizama iz kojih su proizvedeni, te je dan pregled njihove raznovrsne primjene.The use of enzymes or microorganisms in food preparations is an age-old process. With the advancement of technology, novel enzymes with wide range of applications and specificity have been developed and new application areas are still being explored. Microorganisms such as bacteria, yeast and fungi and their enzymes are widely used in several food preparations for improving the taste and texture and they offer huge economic benefits to industries. Microbial enzymes are the preferred source to plants or animals due to several advantages such as easy, cost-effective and consistent production. The present review discusses the recent advancement in enzyme technology for food industries. A comprehensive list of enzymes used in food processing, the microbial source of these enzymes and the wide range of their application are discussed

In vivo genome editing improves muscle function in a mouse model of Duchenne muscular dystrophy

Duchenne muscular dystrophy (DMD) is a devastating disease affecting about 1 out of 5000 male births and caused by mutations in the dystrophin gene. Genome editing has the potential to restore expression of a modified dystrophin gene from the native locus to modulate disease progression. In this study, adeno-associated virus was used to deliver the CRISPR/Cas9 system to the mdx mouse model of DMD to remove the mutated exon 23 from the dystrophin gene. This includes local and systemic delivery to adult mice and systemic delivery to neonatal mice. Exon 23 deletion by CRISPR/Cas9 resulted in expression of the modified dystrophin gene, partial recovery of functional dystrophin protein in skeletal myofibers and cardiac muscle, improvement of muscle biochemistry, and significant enhancement of muscle force. This work establishes CRISPR/Cas9-based genome editing as a potential therapy to treat DMD

Harnessing the probiotic properties and immunomodulatory effects of fermented food-derived Limosilactobacillus fermentum strains: implications for environmental enteropathy

IntroductionEnvironmental enteropathy (EE), a chronic small intestine disease characterized by gut inflammation, is widely prevalent in low-income countries and is hypothesized to be caused by continuous exposure to fecal contamination. Targeted nutritional interventions using potential probiotic strains from fermented foods can be an effective strategy to inhibit enteric pathogens and prevent chronic gut inflammation.MethodsWe isolated potential strains from fermented rice water and lemon pickle and investigated their cell surface properties, antagonistic properties, adhesion to HT-29 cells, and inhibition of pathogen adherence to HT-29 cells. Bacteriocin-like inhibitory substances (BLIS) were purified, and in vivo, survival studies in Caenorhabditis elegans infected with Salmonella enterica MW116733 were performed. We further checked the expression pattern of pro and anti-inflammatory cytokines (IL-6, IL8, and IL-10) in HT-29 cells supplemented with strains.ResultsThe strains isolated from rice water (RS) and lemon pickle (T1) were identified as Limosilactobacillus fermentum MN410703 and MN410702, respectively. Strains showed probiotic properties like tolerance to low pH (pH 3.0), bile salts up to 0.5%, simulated gastric juice at low pH, and binding to extracellular matrix molecules. Auto-aggregation of T1 was in the range of 85% and significantly co-aggregated with Klebsiella pneumoniae, S. enterica, and Escherichia coli at 48, 79, and 65%, respectively. Both strains had a higher binding affinity to gelatin and heparin compared to Bacillus clausii. Susceptibility to most aminoglycoside, cephalosporin, and macrolide classes of antibiotics was also observed. RS showed BLIS activity against K. pneumoniae, S. aureus, and S. enterica at 60, 48, and 30%, respectively, and the protective effects of BLIS from RS in the C. elegans infection model demonstrated a 70% survival rate of the worms infected with S. enterica. RS and T1 demonstrated binding efficiency to HT-29 cell lines in the 38–46% range, and both strains inhibited the adhesion of E. coli MDR and S. enterica. Upregulation of IL-6 and IL-10 and the downregulation of IL-8 were observed when HT-29 cells were treated with RS, indicating the immunomodulatory effects of the strain.DiscussionThe potential strains identified could effectively inhibit enteric pathogens and prevent environmental enteropathy

Micropropagation and conservation of selected endangered anticancer medicinal plants from the Western Ghats of India

Globally, cancer is a constant battle which severely affects the human population. The major limitations of the anticancer drugs are the deleterious side effects on the quality of life. Plants play a vital role in curing many diseases with minimal or no side effects. Phytocompounds derived from various medicinal plants serve as the best source of drugs to treat cancer. The global demand for phytomedicines is mostly reached by the medicinal herbs from the tropical nations of the world even though many plant species are threatened with extinction. India is one of the mega diverse countries of the world due to its ecological habitats, latitudinal variation, and diverse climatic range. Western Ghats of India is one of the most important depositories of endemic herbs. It is found along the stretch of south western part of India and constitutes rain forest with more than 4000 diverse medicinal plant species. In recent times, many of these therapeutically valued herbs have become endangered and are being included under the red-listed plant category in this region. Due to a sharp rise in the demand for plant-based products, this rich collection is diminishing at an alarming rate that eventually triggered dangerous to biodiversity. Thus, conservation of the endangered medicinal plants has become a matter of importance. The conservation by using only in situ approaches may not be sufficient enough to safeguard such a huge bio-resource of endangered medicinal plants. Hence, the use of biotechnological methods would be vital to complement the ex vitro protection programs and help to reestablish endangered plant species. In this backdrop, the key tools of biotechnology that could assist plant conservation were developed in terms of in vitro regeneration, seed banking, DNA storage, pollen storage, germplasm storage, gene bank (field gene banking), tissue bank, and cryopreservation. In this chapter, an attempt has been made to critically review major endangered medicinal plants that possess anticancer compounds and their conservation aspects by integrating various biotechnological tool

Characterizing low-frequency flame-front oscillations – An investigation into their origin and possible mitigation

This investigation focuses on the premixed conical flame, a mainstay of combustion diagnostics. While extensive studies have explored high-frequency oscillations in the canonical Bunsen flame, little attention has been given to low-frequency oscillations affecting flame fronts. This thesis aims to explain the origins of these oscillations and proposes mitigation strategies. The goal is to improve high-accuracy diagnostics of standard conical flames by generating stable flame plumes through oscillation reduction.The thesis explores two theories for low-frequency oscillations: buoyancy variations and flow rate fluctuations. Image chemiluminescence and CARS spectroscopy validate these hypotheses. Spatial oscillations are detected using flame front edge detection algorithms, while CARS analysis indirectly confirms fluctuations. Preliminary experiments establish reference data sets, followed by a primary experimental campaign spanning Reynolds numbers from 500 to 1000 for constant equivalence ratios. This isolates the effect of buoyancy variations. The report concludes by suggesting simple design changes for the flame holder to reduce instability amplitudes.Aerospace Engineerin

Synthetic Biology and Metabolic Engineering Approaches and Its Impact on Non-Conventional Yeast and Biofuel Production

The increasing fossil fuel scarcity has led to an urgent need to develop alternative fuels. Currently microorganisms have been extensively used for the production of first-generation biofuels from lignocellulosic biomass. Yeast is the efficient producer of bioethanol among all existing biofuels option. Tools of synthetic biology have revolutionized the field of microbial cell factories especially in the case of ethanol and fatty acid production. Most of the synthetic biology tools have been developed for the industrial workhorse Saccharomyces cerevisiae. The non-conventional yeast systems have several beneficial traits like ethanol tolerance, thermotolerance, inhibitor tolerance, genetic diversity, etc., and synthetic biology have the power to expand these traits. Currently, synthetic biology is slowly widening to the non-conventional yeasts like Hansenula polymorpha, Kluyveromyces lactis, Pichia pastoris, and Yarrowia lipolytica. Herein, we review the basic synthetic biology tools that can apply to non-conventional yeasts. Furthermore, we discuss the recent advances employed to develop efficient biofuel-producing non-conventional yeast strains by metabolic engineering and synthetic biology with recent examples. Looking forward, future synthetic engineering tools’ development and application should focus on unexplored non-conventional yeast species