AI Article Synopsis

  • - The study focuses on the insect pathogenic fungus responsible for honeybee chalk brood disease and explores the optimization of amylase production using response surface methodology (RSM) and Box-Behnken design (BBD) for enhanced enzyme activity.
  • - The optimized media led to a significant increase in amylase activity (45.28 U/mL) compared to standard media, with the purified enzyme demonstrating ideal conditions of 55 °C and pH 7.5, while showing inhibition by certain metals and solvents.
  • - This research highlights the potential biotechnological applications of the enzyme, especially given its characteristics as an endo-amylase, and presents an effective strategy for producing fungal enzymes on a larger scale.

Article Abstract

The insect pathogenic fungus, , is the causative agent of honeybee chalk brood disease. Amylases are secreted by many plant pathogenic fungi to access host nutrients through the metabolism of starch, and the identification of new amylases can have important biotechnological applications. Production of amylase by in submerged culture was optimized using the response surface method (RSM). Media composition was modeled using Box-Behnken design (BBD) at three levels of three variables, and the model was experimentally validated to predict amylase activity ( = 0.9528). Amylase activity was highest (45.28 ± 1.16 U/mL, mean ± SE) in media composed of 46 g/L maltose and1.51 g/L CaCl at a pH of 6.6, where total activity was ~11-fold greater as compared to standard basal media. The enzyme was purified to homogeneity with a 2.5% yield and 14-fold purification. The purified enzyme had a molecular weight of 75 kDa and was thermostable and active in a broad pH range (> 80% activity at a pH range of 7-10), with optimal activity at 55 °C and pH = 7.5. Kinetic analyses revealed a of 6.22 mmol/L and a of 4.21 μmol/mL·min using soluble starch as the substrate. Activity was significantly stimulated by Fe and completely inhibited by Cu, Mn, and Ba (10 mM). Ethanol and chloroform (10% /) also caused significant levels of inhibition. The purified amylase essentially exhibited activity only on hydrolyzed soluble starch, producing mainly glucose and maltose, indicating that it is an endo-amylase (α-amylase). Amylase activity peaked at 99.38 U/mL fermented in a 3.7 L-bioreactor (2.15-fold greater than what was observed in flask cultures). These data provide a strategy for optimizing the production of enzymes from fungi and provide insight into the α-amylase of .

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10672707PMC
http://dx.doi.org/10.3390/jof9111082DOI Listing

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