Conductive Ink-Coated Paper-Based Supersandwich DNA Biosensor for Ultrasensitive Detection of Neisseria gonorrhoeae.

Herein, we report results of the studies relating to the development of an impedimetric, magnetic bead-assisted supersandwich DNA hybridization assay for ultrasensitive detection of Neisseria gonorrhoeae, the causative agent of a sexually transmitted infection (STI), gonorrhea. First, a conductive ink was formulated by homogenously dispersing carboxylated multiwalled carbon nanotubes (cMWCNTs) in a stable emulsion of terpineol and an aqueous suspension of carboxymethyl cellulose (CMC). The ink, labeled C5, was coated onto paper substrates to fabricate C5@paper conductive electrodes. Thereafter, a magnetic bead (MB)-assisted supersandwich DNA hybridization assay was optimized against the porA pseudogene of N. gonorrhoeae. For this purpose, a pair of specific 5' aminated capture probes (SCP) and supersandwich detector probes (SDP) was designed, which allowed the enrichment of target gonorrheal DNA sequence from a milieu of substances. The SD probe was designed such that instead of 1:1 binding, it allowed the binding of more than one T strand, leading to a 'ladder-like' DNA supersandwich structure. The MB-assisted supersandwich assay was integrated into the C5@paper electrodes for electrochemical analysis. The C5@paper electrodes were found to be highly conductive by a four-probe conductivity method (maximum conductivity of 10.1 S·cm-1). Further, the biosensing assay displayed a wide linear range of 100 aM-100 nM (109 orders of magnitude) with an excellent sensitivity of 22.6 kΩ·(log[concentration])-1. The clinical applicability of the biosensing assay was assessed by detecting genomic DNA extracted from N. gonorrhoeae in the presence of DNA from different non-gonorrheal bacterial species. In conclusion, this study demonstrates a highly sensitive, cost-effective, and label-free paper-based device for STI diagnostics. The ink formulation prepared for the study was found to be highly thixotropic, which indicates that the paper electrodes can be screen-printed in a reproducible and scalable manner.

Recent developments in wearable & non-wearable point-of-care biosensors for cortisol detection.

INTRODUCTION: Cortisol is one of the most prominent biomarkers used for the detection of psychological stress and related disorders. It plays an important role in many physiological processes including immunomodulation and fat metabolism. Thus, monitoring of cortisol levels can be used to indicate different pathological conditions including stress disorders. There has been a gradual rise in the development of point of care (PoC) biosensors for continuous cortisol monitoring. AREAS COVERED: This review discusses recent breakthroughs toward the development of PoC sensors (wearable and non wearable) for cortisol monitoring. Challenges associated with them have also been summarized. EXPERT OPINION: Electrochemical PoC devices have recently emerged as a powerful tools for continuous monitoring of cortisol that can be utilized for stress management and treatment of related disorders. However, there are many challenges that should be addressed before such devices can be deployed at mass level, such as inter-individual variability, changing the device calibration with the circadian rhythm, interference from other endocrine moieties, etc. [Figure: see text].

Electrochemical Immunosensor for Detection of H. pylori Secretory Protein VacA on g-C3N4/ZnO Nanocomposite-Modified Au Electrode.

A g-C3N4/ZnO (graphitic carbon nitride/zinc oxide) nanocomposite-decorated gold electrode was employed to design an antigen-antibody-based electrochemical biosensor to detect Helicobacter pylori specific toxin, vacuolating cytotoxin A (VacA). The thermal condensation method was used to synthesize the g-C3N4/ZnO nanocomposite, and the nanocomposite was deposited electrochemically on a gold electrode. The morphology as well as the structure of the synthesized nanocomposite were confirmed by scanning electron microscopy, energy-dispersive X-ray analysis, X-ray diffraction, and Fourier transform infrared techniques. The nanocomposite efficiently increased the sensor performance by amplifying the signals. EDC-NHS chemistry was exploited for attachment of VacA antibodies covalently with the g-C3N4/ZnO-modified gold electrode. This modified electrode was exploited for immunosensing of H. pylori-specific VacA antigen. The immunosensor was stable for up to 30 days and exhibited good sensitivity of 0.3 µA-1 ng mL-1 in a linear detection range of 0.1 to 12.8 ng mL-1. Apart from this, the fabricated sensor showed unprecedented reproducibility and remarkable selectivity toward the H. pylori toxin VacA. Thus, the highly sensitive immunosensor is a desirable platform for H. pylori detection in practical applications and clinical diagnosis.

Applications of metal-organic framework-based bioelectrodes.

Metal-organic frameworks (MOFs) are an emerging class of porous nanomaterials that have opened new research possibilities. The inherent characteristics of MOFs such as their large surface area, high porosity, tunable pore size, stability, facile synthetic strategies and catalytic nature have made them promising materials for enormous number of applications, including fuel storage, energy conversion, separation, and gas purification. Recently, their high potential as ideal platforms for biomolecule immobilization has been discovered. MOF-enzyme-based materials have attracted the attention of researchers from all fields with the expansion of MOFs development, paving way for the fabrication of bioelectrochemical devices with unique characteristics. MOFs-based bioelectrodes have steadily gained interest, wherein MOFs can be utilized for improved biomolecule immobilization, electrolyte membranes, fuel storage, biocatalysis and biosensing. Likewise, applications of MOFs in point-of-care diagnostics, including self-powered biosensors, are exponentially increasing. This paper reviews the current trends in the fabrication of MOFs-based bioelectrodes with emphasis on their applications in biosensors and biofuel cells.

Point-of-Care PCR Assays for COVID-19 Detection.

Molecular diagnostics has been the front runner in the world's response to the COVID-19 pandemic. Particularly, reverse transcriptase-polymerase chain reaction (RT-PCR) and the quantitative variant (qRT-PCR) have been the gold standard for COVID-19 diagnosis. However, faster antigen tests and other point-of-care (POC) devices have also played a significant role in containing the spread of SARS-CoV-2 by facilitating mass screening and delivering results in less time. Thus, despite the higher sensitivity and specificity of the RT-PCR assays, the impact of POC tests cannot be ignored. As a consequence, there has been an increased interest in the development of miniaturized, high-throughput, and automated PCR systems, many of which can be used at point-of-care. This review summarizes the recent advances in the development of miniaturized PCR systems with an emphasis on COVID-19 detection. The distinct features of digital PCR and electrochemical PCR are detailed along with the challenges. The potential of CRISPR/Cas technology for POC diagnostics is also highlighted. Commercial RT-PCR POC systems approved by various agencies for COVID-19 detection are discussed.

Emerging DNA-based multifunctional nano-biomaterials towards electrochemical sensing applications.

DNA is known to be ubiquitous in nature as it is the controlling unit for genetic information storage in most living organisms. Lately, there has been a surge in studies relating to the use of DNA as a biomaterial for various biomedical applications such as biosensing, therapeutics, and drug delivery. The role of DNA as a bioreceptor in biosensors has been known for a long time. DNA-based biosensors are gradually evolving into highly sophisticated and sensitive molecular devices. The current realization of DNA-based biosensors embraces the unique structural and functional properties of DNA in the form of a biopolymer. The interesting properties of DNA, such as self-assembly, programmability, catalytic activity, dynamic behavior, and precise molecular recognition, have led to the emergence of innovative DNA assembly based electrochemical biosensors. This review article aims to cover the recent progress in the field of DNA-based electrochemical (EC) biosensors. It commences with an introduction to electrochemical biosensors and elucidates the advantages of integrating DNA-based materials into them. Besides this, we discuss the principles of EC biosensors based on different types of DNA-based materials. The article concludes by highlighting the outlook and importance of this interesting field for biomedical developments.

Ultrasensitive biosensing platform based on yttria doped zirconia-reduced graphene oxide nanocomposite for detection of salivary oral cancer biomarker.

Herein, we report results of the studies relating to the fabrication of yttria-doped zirconia-reduced graphene oxide nanocomposite (nYZR) based biosensing platform for detection of salivary CYFRA-21-1 biomarker. The nYZR nanocomposite was hydrothermally synthesized and amine-functionalized using 3-aminopropyl triethoxysilane (APTES). This functionalized nanocomposite (APTES/nYZR) was electrophoretically deposited (45 V; 3 min) onto pre-hydrolyzed indium tin oxide (ITO) coated glass substrate (APTES/nYZR/ITO) followed by biofunctionalization via covalent immobilization of the anti-CYFRA-21-1 antibodies (anti-CYFRA-21-1/APTES/nYZR/ITO). The synthesized nanomaterial and the fabricated electrodes were characterized to investigate crystal structure, morphology and electrochemical properties via X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy. The fabricated biosensing electrode (BSA/anti-CYFRA-21-1/APTES/nYZR/ITO) has an operating shelf life of 56 days and can be used to detect salivary CYFRA-21-1 biomarker concentration as low as 7.2 pg mL-1 with wide linear detection range of 0.01-50 ng mL-1. This work opens new opportunities to explore the electrochemical behavior of nanostructured yttria stabilized zirconia (YSZ) and its composites at room temperature and its utility in developing biosensors and other electrochemical devices.

Gold nanomaterials for optical biosensing and bioimaging.

Gold nanoparticles (AuNPs) are highly compelling nanomaterials for biomedical studies due to their unique optical properties. By leveraging the versatile optical properties of different gold nanostructures, the performance of biosensing and biomedical imaging can be dramatically improved in terms of their sensitivity, specificity, speed, contrast, resolution and penetration depth. Here we review recent advances of optical biosensing and bioimaging techniques based on three major optical properties of AuNPs: surface plasmon resonance, surface enhanced Raman scattering and luminescence. We summarize the fabrication methods and optical properties of different types of AuNPs, highlight the emerging applications of these AuNPs for novel optical biosensors and biomedical imaging innovations, and discuss the future trends of AuNP-based optical biosensors and bioimaging as well as the challenges of implementing these techniques in preclinical and clinical investigations.

Prospects of nanomaterials-enabled biosensors for COVID-19 detection.

We are currently facing the COVID-19 pandemic which is the consequence of severe acute respiratory syndrome coronavirus (SARS-CoV-2). Since no specific vaccines or drugs have been developed till date for the treatment of SARS-CoV-2 infection, early diagnosis is essential to further combat this pandemic. In this context, the reliable, rapid, and low-cost technique for SARS-CoV-2 diagnosis is the foremost priority. At present reverse transcription polymerase chain reaction (RT-PCR) is the reference technique presently being used for the detection of SARS-CoV-2 infection. However, in a number of cases, false results have been noticed in COVID-19 diagnosis. To develop advanced techniques, researchers are continuously working and in the series of constant efforts, nanomaterials-enabled biosensing approaches can be a hope to offer novel techniques that may perhaps meet the current demand of fast and early diagnosis of COVID-19 cases. This paper provides an overview of the COVID-19 pandemic and nanomaterials-enabled biosensing approaches that have been recently reported for the diagnosis of SARS-CoV-2. Though limited studies on the development of nanomaterials enabled biosensing techniques for the diagnosis of SARS-CoV-2 have been reported, this review summarizes nanomaterials mediated improved biosensing strategies and the possible mechanisms that may be responsible for the diagnosis of the COVID-19 disease. It is reviewed that nanomaterials e.g. gold nanostructures, lanthanide-doped polysterene nanoparticles (NPs), graphene and iron oxide NPs can be potentially used to develop advanced techniques offered by colorimetric, amperometric, impedimetric, fluorescence, and optomagnetic based biosensing of SARS-CoV-2. Finally, critical issues that are likely to accelerate the development of nanomaterials-enabled biosensing for SARS-CoV-2 infection have been discussed in detail. This review may serve as a guide for the development of advanced techniques for nanomaterials enabled biosensing to fulfill the present demand of low-cost, rapid and early diagnosis of COVID-19 infection.

Recent Advances of Conducting Polymers and Their Composites for Electrochemical Biosensing Applications.

Conducting polymers (CPs) have been at the center of research owing to their metal-like electrochemical properties and polymer-like dispersion nature. CPs and their composites serve as ideal functional materials for diversified biomedical applications like drug delivery, tissue engineering, and diagnostics. There have also been numerous biosensing platforms based on polyaniline (PANI), polypyrrole (PPY), polythiophene (PTP), and their composites. Based on their unique properties and extensive use in biosensing matrices, updated information on novel CPs and their role is appealing. This review focuses on the properties and performance of biosensing matrices based on CPs reported in the last three years. The salient features of CPs like PANI, PPY, PTP, and their composites with nanoparticles, carbon materials, etc. are outlined along with respective examples. A description of mediator conjugated biosensor designs and enzymeless CPs based glucose sensing has also been included. The future research trends with required improvements to improve the analytical performance of CP-biosensing devices have also been addressed.

Emerging Trends in Microfluidics Based Devices.

One of the major challenges for scientists and engineers today is to develop technologies for the improvement of human health in both developed and developing countries. However, the need for cost-effective, high-performance diagnostic techniques is very crucial for providing accessible, affordable, and high-quality healthcare devices. In this context, microfluidic-based devices (MFDs) offer powerful platforms for automation and integration of complex tasks onto a single chip. The distinct advantage of MFDs lies in precise control of the sample quantities and flow rate of samples and reagents that enable quantification and detection of analytes with high resolution and sensitivity. With these excellent properties, microfluidics (MFs) have been used for various applications in healthcare, along with other biological and medical areas. This review focuses on the emerging demands of MFs in different fields such as biomedical diagnostics, environmental analysis, food and agriculture research, etc., in the last three or so years. It also aims to reveal new opportunities in these areas and future prospects of commercial MFDs.

Nanomaterial-Modified Conducting Paper: Fabrication, Properties, and Emerging Biomedical Applications.

The emerging demand for wearable, lightweight portable devices has led to the development of new materials for flexible electronics using non-rigid substrates. In this context, nanomaterial-modified conducting paper (CP) represents a new concept that utilizes paper as a functional part in various devices. Paper has drawn significant interest among the research community because it is ubiquitous, cheap, and environmentally friendly. This review provides information on the basic characteristics of paper and its functionalization with nanomaterials, methodology for device fabrication, and their various applications. It also highlights some of the exciting applications of CP in point-of-care diagnostics for biomedical applications. Furthermore, recent challenges and opportunities in paper-based devices are summarized.

Biofunctionalized Nanostructured Yttria Modified Non-Invasive Impedometric Biosensor for Efficient Detection of Oral Cancer.

We report results of the studies relating to the development of an efficient biosensor for non-invasive detection of CYFRA-21-1 cancer biomarker. We used a low dielectric constant material (nanostructured yttrium oxide, nY2O3) for the fabrication of the biosensing platform. The nY2O3 was synthesized via solvothermal process and functionalized using 3-aminopropyl triethoxy silane (APTES). Electrophoretic deposition (EPD) of the functionalized nanomaterial (APTES/nY2O3) onto an indium tin oxide (ITO)-coated glass electrode was conducted at a DC potential of 50V for 60s. The EDC-NHS chemistry was used for covalent immobilization of -COOH bearing monoclonal anti-CYFRA-21-1 onto -NH2 groups of APTES/nY2O3/ITO electrode. To avoid the non-specific interaction on the anti-CYFRA-21-1/APTES/nY2O3/ITO immunoelectrode, bovine serum albumin (BSA) was used. X-ray diffraction (XRD), transmission electron microscopy (TEM), and field emission scanning electron microscopy (FESEM) were utilized for structural and morphological studies, whereas Fourier-transform infrared spectroscopy (FTIR) was used for the bonding analysis. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques were used for electrochemical characterization and response studies of fabricated electrodes. The fabricated immunosensor (BSA/anti-CYFRA-21-1/APTES/nY2O3/ITO) exhibited linearity in the range of 0.01-50 ng·mL-1, sensitivity of 226.0 Ω·mL·ng-1, and lower detection limit of 0.01·ng·mL-1. A reasonable correlation was observed between the results obtained using this biosensor and concentration of CYFRA-21-1 measured through ELISA (enzyme-linked immunosorbent assay) technique in salivary samples of oral cancer patients.

Cell-based biosensors: Recent trends, challenges and future perspectives.

Existing at the interface of biology and electronics, living cells have been in use as biorecognition elements (bioreceptors) in biosensors since the early 1970s. They are an interesting choice of bioreceptors as they allow flexibility in determining the sensing strategy, are cheaper than purified enzymes and antibodies and make the fabrication relatively simple and cost-effective. And with advances in the field of synthetic biology, microfluidics and lithography, many exciting developments have been made in the design of cell-based biosensors in the last about five years. 3D cell culture systems integrated with electrodes are now providing new insights into disease pathogenesis and physiology, while cardiomyocyte-integrated microelectrode array (MEA) technology is set to be standardized for the assessment of drug-induced cardiac toxicity. From cell microarrays for high-throughput applications to plasmonic devices for anti-microbial susceptibility testing and advent of microbial fuel cell biosensors, cell-based biosensors have evolved from being mere tools for detection of specific analytes to multi-parametric devices for real time monitoring and assessment. However, despite these advancements, challenges such as regeneration and storage life, heterogeneity in cell populations, high interference and high costs due to accessory instrumentation need to be addressed before the full potential of cell-based biosensors can be realized at a larger scale. This review summarizes results of the studies that have been conducted in the last five years toward the fabrication of cell-based biosensors for different applications with a comprehensive discussion on the challenges, future trends, and potential inputs needed for improving them.

Protein functionalised self assembled monolayer based biosensor for colon cancer detection.

We report results of the studies relating to the fabrication of a surface plasmon resonance (SPR) based label-free immunosensor for real-time monitoring of endothelin-1 (ET-1), a colon cancer biomarker. A gold disk modified with a self-assembled monolayer (SAM) of 11-mercaptoundecanoic acid (11-MUA) was functionalised via covalent immobilization of monoclonal anti-ET-1 antibodies using EDC-NHS (1-(3-(dimethylamine)-propyl)-3-ethylcarbodiimide hydrochloride, N-hydroxy succinimide) chemistry. This immunosensing platform (ethanolamine/anti-ET-1/11-MUA/Au) was characterized via atomic force microscopy (AFM), contact angle (CA) and Fourier transform infrared (FT-IR) spectroscopic techniques. The fabricated SPR electrode was further used to detect ET-1 in the broad concentration range 2-100 pg mL-1, with a detection limit of 0.30 pg mL-1 and remarkable sensitivity of 2.18 mo pg-1mL. The adsorption mechanism was studied using monophasic model and the values of association (ka) and dissociation (kd) constants for anti-ET-1 and ET-1 binding were calculated to be 4.4 ±â€¯0.4 × 105 M-1 s-1 and 2.04 ±â€¯0.0003 × 10-3 s-1, respectively. The results obtained via analysis of serum samples of colorectal cancer patients were found to be in good agreement with those obtained from enzyme-linked immunosorbent assay (ELISA) technique. Further, electrochemical studies were performed to prove the efficacy of the fabricated platform as a point of care device for the detection of ET-1.

Nanoengineered cellulosic biohydrogen production via dark fermentation: A novel approach.

The insights of nanotechnology for cellulosic biohydrogen production through dark fermentation are reviewed. Lignocellulosic biomass to sugar generation is a complex process and covers the most expensive part of cellulose to sugar production technology. In this context, the impacts of nanomaterial on lignocellulosic biomass to biohydrogen production process have been reviewed. In addition, the feasibility of nanomaterials for implementation in each step of the cellulosic biohydrogen production is discussed for economic viability of the process. Numerous aspects such as possible replacement of chemical pretreatment method using nanostructured materials, use of immobilized enzyme for a fast rate of reaction and its reusability along with long viability of microbial cells and hydrogenase enzyme for improving the productivity are the highlights of this review. It is found that various types of nanostructured materials e.g. metallic nanoparticles (Fe°, Ni, Cu, Au, Pd, Au), metal oxide nanoparticles (Fe2O3, F3O4, NiCo2O4, CuO, NiO, CoO, ZnO), nanocomposites (Si@CoFe2O4, Fe3O4/alginate) and graphene-based nanomaterials can influence different parameters of the process and therefore may perhaps be utilized for cellulosic biohydrogen production. The emphasis has been given on the cost issue and synthesis sustainability of nanomaterials for making the biohydrogen technology cost effective. Finally, recent advancements and feasibility of nanomaterials as the potential solution for improved cellulose conversion to the biohydrogen production process have been discussed, and this is likely to assist in developing an efficient, economical and sustainable biohydrogen production technology.

Electrochemical paper based cancer biosensor using iron oxide nanoparticles decorated PEDOT:PSS.

We report results of the studies relating to the fabrication of a label-free, flexible, light weight and disposable conducting paper based immunosensing platform comprising of poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) and nanostructured iron oxide (nFe2O3@PEDOT:PSS) nanocomposite for detection of carcinoembryonic antigen (CEA), a cancer biomarker. The effect of various solvents such as sorbitol, ethanol, propanol, n-methyl-2-pyrrolidone (NMP) and dimethyl sulfoxide (DMSO) on the electrical conductivity of Whatman filter paper (WP) modified with nFe2O3@PEDOT:PSS/WP was investigated. The electrical conductivity of the PEDOT:PSS/WP electrode was found to be enhanced by two orders of magnitude (from 6.8× 10-4 to 1.92 × 10-2 Scm-1) after its treatment with DMSO. Further, nFe2O3 doped PEDOT:PSS/WP electrode exhibited the electrical conductivity as 2.4 × 10-2 Scm-1. Besides this, the incorporation of iron oxide nanoparticles (nFe2O3) into PEDOT:PSS/WP resulted in improved electrochemical performance and signal stability. This nFe2O3@PEDOT:PSS/WP based platform was used for immobilization of the anti-carcinoembronic antigen (anti-CEA) protein for quantitative estimation of cancer biomarker (CEA). The results of electrochemical response studies revealed that this conducting paper based immunoelectrode had a sensitivity of 10.2 µAng-1mLcm-2 in the physiological range (4-25 ngmL-1) and shelf life of 34 days. Further, the proposed immunoelectrode was validated with conventional ELISA for the detection of CEA in serum samples of cancer patients.

Amine-Functionalized MoO3@RGO Nanohybrid-Based Biosensor for Breast Cancer Detection.

We report results of studies relating to development of an ultrasensitive, rapid, and label-free biosensor based on molybdenum trioxide (MoO3) anchored onto the reduced graphene oxide (RGO) for breast cancer detection. The human epidermal growth factor receptor-2 (HER-2) secreted in the serum of breast cancer patients was used as an analyte for the detection. The in situ growth of 1D MoO3 onto reduced graphene oxide (RGO), a 2D carbon substrate, was carried out via one-pot low-temperature hydrothermal synthesis. Subsequently, the surface conjugation of the monoclonal antibodies (anti-HER-2) onto the APTES/MoO3@RGO/ITO electrode was conducted via EDC-NHS covalent chemistry. The structural and morphological properties of the MoO3@RGO nanohybrid were investigated using electron microscopy, X-ray diffraction, scanning electron microscopy, transmission electron microscopy and X-ray photoelectron spectroscopic techniques. The surface area of the MoO3@RGO nanohybrid determined via Brauner-Emmett-Teller analysis was found to be 14 times greater than that of the pristine MoO3. The binding kinetics and the electrochemical activity of the developed platform were determined by cyclic voltammetry, differential pulse voltammetry, and impedance spectroscopic techniques. This nanohybrid based immunosensor exhibited improved sensitivity (13 uA mLng-1cm-2) in a broad concentration range (0.001-500 ng mL-1) with a correlation coefficient of 0.98. The limit of detection of this MoO3@RGO nanohybrid based immunosensor was found to be 0.001 ng mL-1. The results obtained via the developed immunosensor for the quantification of serum HER-2 were validated using ELISA.

Microfluidics Based Point-of-Care Diagnostics.

Point-of-care (POC) diagnostic devices have been predicted to provide a boon in health care especially in the diagnosis and detection of diseases. POC devices have been found to have many advantages like a rapid and precise response, portability, low cost, and non-requirement of specialized equipment. The major objective of a POC diagnostic research is to develop a chip-based, self-containing miniaturized device that can be used to examine different analytes in complex samples. Further, the integration of microfluidics (MF) with advanced biosensor technologies is likely to result in improved POC diagnostics. This paper presents the overview of the different materials (glass, silicon, polymer, paper) and techniques for the fabrication of MF based POC devices along with their wide range of biosensor applications. Besides this, the authors have presented in brief the challenges that MF is currently facing along with possible solutions that may result in the availability of the accessible, reliable, and cost-efficient technology. The development of these devices requires the combination of developed MF components into POC devices that are user-friendly, sensitive, stable, accurate, low cost, and minimally invasive. These MF based POC devices have tremendous potential in providing improved healthcare including easy monitoring, early detection of disease, and increased personalization.

Production and Optimization of Physicochemical Parameters of Cellulase Using Untreated Orange Waste by Newly Isolated Emericella variecolor NS3.

Cellulase enzymes have versatile industrial applications. This study was directed towards the isolation, production, and characterization of cellulase enzyme system. Among the five isolated fungal cultures, Emericella variecolor NS3 showed maximum cellulase production using untreated orange peel waste as substrate using solid-state fermentation (SSF). Maximum enzyme production of 31 IU/gds (per gram of dry substrate) was noticed at 6.0 g concentration of orange peel. Further, 50 °C was recorded as the optimum temperature for cellulase activity and the thermal stability for 240 min was observed at this temperature. In addition, the crude enzyme was stable at pH 5.0 and held its complete relative activity in presence of Mn2+ and Fe3+. This study explored the production of crude enzyme system using biological waste with future potential for research and industrial applications.