Purification and characterization of Alanine dehydrogenase from Streptomyces anulatus for its application as a bioreceptor in biosensor

Characterization of Alanine Dehydrogenase and Its Effect on <b><i>Streptomyces coelicolor</i></b>A3(2) Development in Liquid Culture

“…Because of the use of purified ScALD enzyme we were able to report all kinetic parameters in meaningful kinetic units unlike specific activity units reported previously due to the use of crude enzyme. We have shown that the enzyme has a high pH optimum, substrate specificity, and temperature profiles for both oxidative deamination and reductive amination reactions, similar to other amino ALD enzymes from different bacterial species, reflecting a common reaction mechanism [Dave and Kadeppagari, 2018;Giffin et al, 2012;Ohashima and Soda, 1979;Phogosee et al, 2018]. For oxidative deamination reaction, the enzyme is highly specific to Lalanine compared to other L-amino donor substrates we have tested.…”

“…This characterization effort adds to the knowledge of the growing interest for ALD in many biotechnological applications including biosensor, biocatalyst, and antimicrobial drug development [Dave and Kadeppagari, 2018; Dave and Kadeppagari, 2019;Hasan et al, 2006]. We also expect that the ScALD can be utilized as an ideal model system for further protein engineering studies to obtain ALDs with improved functional and catalytic properties for different biotechnological applications.…”

“…The oxidative deamination reaction catalyzed by ALD is required for microorganisms to utilize alanine as a nitrogen source [Lin et al, 2012;Salerno et al, 2013]. Since the demonstration of the involvement of ALD for the β-lactam antibiotic production in S. clavuligerus [Aharonowitz and Friedrich, 1980], several Streptomyces species have been analyzed for ALD activities [Dave and Kadeppagari, 2018;Itoh and Morikawa, 1983;Vancurova et al, 1989]. However, as per our knowledge, all enzymatic characterizations were carried out with crude ALD purified from Streptomyces species.…”

Forward-Looking Roadmaps for Long-Term Continuous Water Quality Monitoring: Bottlenecks, Innovations, and Prospects in a Critical Review

“…Biosensors measure the target analyte qualitatively or quantitatively by converting the observed responses into the measurable electrical or optical signals, which is carried out by the biological recognition elements such as enzymes and microorganisms (Figure j). Biosensors have gained enormous attention in the past decade, especially for monitoring nutrients and toxic analytes such as biological oxygen demand (BOD), microbes, nitrogen, pathogens, and heavy metals with the response time of seconds (Figure k, Table ). Microbial fuel cell-based and biofilm-based sensors were developed for monitoring contaminants in WW and demonstrated distinct features such as self-sustained operational mode, small reagent volume (<30 mL , ), low cost (intrinsic microorganisms as recognition elements), − and clear correlations between potential (mV) output and contaminant concentration.…”