Industrial Biotechnology

CRISPR-Cas9 is a groundbreaking technology that has revolutionized genetic engineering by allowing precise and efficient genome editing. Despite its potential, the complexity and variability of genomic data present significant challenges. Artificial intelligence (AI) offers powerful tools to enhance CRISPR-Cas9 applications by improving target identification, minimizing off-target effects, optimizing components, and streamlining data analysis. AI tools have significantly enhanced the CRISPR-Cas9 genomic editing technique in precision agriculture, particularly in the area of disease detection and trait improvement. This review explores the various roles that AI plays in advancing CRISPR-Cas9 technology, highlighting current tools and future directions. This review also explores various AI-driven tools and methods that have been developed to optimize CRISPR-Cas9 genomic editing, from target identification to data analysis, off-target prediction, optimizing the efficiency, accuracy, and safety of gene editing, data analysis, and personalized medicine.

Expression of recombinant proteins is highly dependent on the micro-environment provided to the cells during the cell culture process. Typically, the cell culture process at industrial scale lasts for 7–15 days, and maintaining a suitable nutrient-rich micro-environment is extremely critical to achieve the desired quality of the protein. Composition of Media and feed, and their application in the process, is crucial as they decide the quality and quantity of the protein. Most of the commercially available media and feeds come with proprietary label from the supplier, so there is always an uncertainty on the concentration and time of addition during the process, and hence the variability in desired quality is likely, and it is always the case that we are under-feeding few and over-feeding few other nutrients. In general, the media and feeds complex mixture of nutrients, which are required for cell growth, is primarily made up of amino acids, vitamins, trace metals, etc. Understanding the composition of these components in media and their consumption during the cell culture process can significantly help in directing the outcome of the process in a desired manner. In this work, the focus was to make a single high-throughput high-performance liquid chromatography method for profiling of nine water-soluble vitamins (vitamins B and C) with baseline separation and to develop a robust sample preparation procedure to minimize the matrix interference of other components present in the cell culture media. The outcome of the study achieved both objectives, where all nine water-soluble vitamins were obtained as baseline separated, and the same method could detect the presence of vitamins in cell culture components where no baseline drift was observed, which confirmed the development of robust sample preparation.

Nanotechnology involves the study of matter at the nanoscale level. Significant developments in nanotechnology have drawn a lot of interest in a range of biomedical applications. Synthesis of Syzygium cumini copper nanoparticles (SC-CuNPs) using S. cumini seed extract was primarily confirmed by the color change where light brown turns into dark brown, with sharp absorption maxima at 360 nm. Fourier Transform Infrared Spectroscopy revealed the various functional groups corresponding to the secondary metabolites that are responsible for the stabilization of SC-CuNPs, while X-ray diffraction analysis indicated a crystalline structure with an average particle size of 80 nm. SC-CuNPs were spherical, as revealed by the Scanning Electron Microscope analysis. At 125 μg/mL concentration, the SC-CuNPs exhibited significant antioxidant activity, as evidenced by 53% of hydrogen peroxide scavenging and 55% of free radical scavenging activity as measured by the phosphomolybdenum method. The antibacterial activity of SC-CuNPs revealed maximum zone of inhibition against Escherichia coli (28 ± 0.97 mm) and Staphylococcus aureus (32 ± 0.89 mm). Furthermore, SC-CuNPs demonstrated 43% anti-inflammatory activity at a 500 μg/mL concentration. The current work concludes by emphasizing the non-toxic and cost-effective properties of SC-CuNPs. Additionally, it presents an alluring and promising avenue for future applications in the biomedical field.

In this study, we compare the effectiveness of two distinct resins for the purification of lactoferrin (LF) from skimmed milk and fermentation broth. The first resin, a highly porous and soft agarose-based material, exhibits strong hydrophilic properties, while the second resin, composed of rigid polymethacrylate, is more hydrophobic. The agarose-based resin, SP Seplife 6AG XL 200, demonstrates superior performance in purifying bovine lactoferrin (bLF) from milk and whey. Its hydrophilic structure minimizes undesired hydrophobic interactions, achieving a purity of 97% and a yield of 90%. Notably, the agarose resin effectively isolates lactoferrin despite its low concentration (<0.3%) and the presence of a similar protein, lactoperoxidase, which tends to coelute. Conversely, for the purification of recombinant bLF (rbLF) from the fermentation broth, the Seplife LXPM SP 5504C resin outperforms in both yield and purity, reaching 99% and 98%, respectively. This resin facilitates elution using only 0.5 M NaCl—an improvement over the 0.7 M NaCl required for the agarose resin. Both agarose and methacrylic resins excel at purifying feeds with high concentrations of rbLF (>10 mg/mL), while consistently maintaining excellent purity levels. Both resins are designed to be cost-effective and scalable and can be used in different chromatography systems employing axial or radial flow columns. Overall, the choice of resin significantly influences the purity and yield of lactoferrin, highlighting the importance of resin selection based on the source of protein.

Electric bacteria possess unique electrochemical capabilities, allowing them to transfer electrons to or from solid conductive materials. This ability potentiates the use of electric bacteria in microbial fuel cells (MFCs) to serve as bioelectrochemical systems for generating electricity from organic matters. A more efficient way of producing such energy could be used to create small, independent power sources. While integration of electric bacteria in electric cars is still in its early stages, it offers several potentials for the future of electric cars. Possible strategies to implement the use of electric bacteria in electric cars could include engineering electrically conductive pili (e-PNs) genes as an artificial operon into nonpathogenic Escherichia coli, using strong promoters to optimize gene expression and enhance current output in MFCs. When scaled to an industrially stacked spiral electrode with 100 m² surface area, it can generate up to 6 kW of continuous power. Furthermore, inclusion of heat-tolerating genes can provide a robust use of electric bacteria; this is in addition to quorum sensing molecules to maintain resilience and stability of the biofilm. The design of a continuous growth system can provide long-term duration of bacterial functionality. These applications could also potentially reduce greenhouse gas emissions, lower the carbon footprint, and contribute to a more sustainable transportation ecosystem. While the use of electric bacteria is still evolving, it holds promise as a sustainable and eco-friendly energy source.