1.Optimizing electrochemical DNA biosensors for the detection of avian infectious bronchitis virus
Md. Safiul Alam Bhuiyan ; Gilbert Ringgit ; Zarina Amin ; Ag Muhammad Sagaf Abu Bakar ; Suryani Saallah ; Sharifudin Md. Shaarani ; Shafiquzzaman Siddiquee
Malaysian Journal of Microbiology 2023;19(no.6):651-663
Aims:
The aim of the study is to develop the optimised parameters of electrochemical DNA biosensors for the specific detection of the Infectious Bronchitis Virus (IBV) in chickens. The goal is to further create a highly sensitive and specific biosensor that can be used for on-site monitoring of IBV on poultry farms.
Methodology and results:
In this study, an electrochemical DNA biosensor was developed for detecting a specific sequence in the IBV genome. The process involved attaching a NH2-ssDNA probe to a gold electrode, followed by
hybridization with the target DNA. Various parameters like buffer, pH, scan rate, incubation time, redox indicators and temperature were optimised using cyclic voltammetry. The probe DNA was designed to enhance hybridization efficiency, which was assessed by measuring current signals. The biosensor, under optimal conditions, demonstrated high sensitivity and specificity when tested with different sequences, including complementary, non-complementary and mismatched ones. Cross-reactivity studies against non-IBV viruses showed distinguishable current signals. These findings have implications for developing a portable on-site IBV monitoring device for use on farms.
Conclusion, significance and impact of study
The optimised parameters and specificity of the electrochemical DNA biosensor suggest its potential for the development of a portable device for on-site monitoring of IBV on poultry farms. This device could prove to be a valuable tool for the early detection of IBV, helping to prevent further spread of the disease. However, it's essential to conduct further research to ensure the practicality and accuracy of the biosensor in real-world farm settings.
2.Enhancing stability and activity of β-galactosidase from Kluyveromyces lactis through immobilization on polymethacrylate monolith and optimisation using response surface methodology
Rima Fatira Dahari ; Mailin Misson ; Clarence M Ongkudon ; Suryani Saallah ; Aliyah Madihah Asran ; Daniel Joe Dailin ; Lai Fatt Chuah ; Hesham El Enshasy
Malaysian Journal of Microbiology 2023;19(no.6):752-765
Aims:
This study investigates the potential of porous polymethacrylate monoliths as enzyme support materials for large-scale enzyme commercialization.
Methodology and results:
It focuses on their preparation and various immobilization techniques, such as adsorption, covalent-binding and cross-linking, specifically applied to β-galactosidase for bioprocess applications. The research assesses immobilization performance, operational stability, reusability and optimization using response surface methodology (RSM). The results reveal that covalent-binding exhibited the highest enzyme activity recovery, while cross-linking showed superior performance at lower enzyme concentrations but decreased at higher concentrations. Covalent-bound enzymes demonstrated reusability for up to four cycles, with optimal pH ranging between 7 and 8 and optimal temperature ranging between 30 °C and 40 °C. Furthermore, RSM optimization highlighted the significant influence of substrate concentration on enzyme activity, with a reliable model (R2 = 0.9163) and adequate precision (S/N = 13.1409).
Conclusion, significance and impact of study
Overall, this study provides valuable guidelines for effectively employing porous monoliths in large-scale industrial bioprocessing, offering potential cost-saving benefits and enhanced efficiency in enzyme commercialization.