Purifikacija i svojstva kolagenolitičke proteaze proizvedene s pomoću Bacillus cereus MBL13

A novel collagenase-producing bacterium has been isolated and identified as Bacillus cereus MBL13. From the culture supernatant of B. cereus MBL13 grown on bone collagen as the sole carbon and nitrogen source, an extracellular protease with novel property of hydrolyzing waste animal bones was purified. The molecular mass of the purified collagenolytic protease was estimated to be (38.0±1.5) kDa. As determined by amino acid analysis, it had high contents of asparagine, lysine and serine. The optimum temperature and pH for the collagenase activity were 40 °C and pH=8.0, respectively. The results of the effects of some metal ions, inhibitors and protein substrates suggested that the purified collagenolytic protease is a member of the metalloproteases. Type I collagen (the typical collagen in animal bone) was used as the substrate for determination of Michaelis-Menten kinetics. The obtained Km value was (1.31±0.05) g/L and the corresponding vmax value was (12.54±2.5) μmol/min. The study assumes that the collagenolytic protease purified from B. cereus MBL13 strain could be applied in the hydrolysis of waste animal bones.Izolirana je nova bakterija koja proizvodi kolagenazu, identificirana kao Bacillus cereus MBL13. Iz supernatanta kulture B. cereus MBL13, uzgojene na koštanom kolagenu kao izvoru ugljika i dušika, izdvojena je ekstracelularna proteaza sa sposobnošću razgradnje životinjskih kostiju. Procijenjena je molekularna masa pročišćene kolagenolitičke proteaze od (38,0±1,5) kDa. Analiziran je aminokiselinski sastav i utvrđeno da enzim sadržava veliku količinu asparagina, lizina i serina. Optimalna temperatura za aktivnost kolagenaze bila je 40 °C, a optimalna pH-vrijednost 8,0. Prema utjecaju iona metala, inhibitora i proteinskih supstrata na aktivnost enzima zaključeno je da enzim pripada u grupu metaloproteaza. Kolagen tipa I (tipični sastojak životinjskih kostiju) upotrijebljen je kao supstrat pri određivanju parametara Michaelis-Mentenove kinetike. Km vrijednost bila je (1,31±0,05) g/L, a vmax (12,54±2,5) μmol/min. Zaključeno je da se kolagenolitička proteaza izdvojena iz B. cereus MBL13 može upotrijebiti za razgradnju životinjskih kostiju

Purifikacija i svojstva kolagenolitičke proteaze proizvedene s pomoću Bacillus cereus MBL13

A novel collagenase-producing bacterium has been isolated and identified as Bacillus cereus MBL13. From the culture supernatant of B. cereus MBL13 grown on bone collagen as the sole carbon and nitrogen source, an extracellular protease with novel property of hydrolyzing waste animal bones was purified. The molecular mass of the purified collagenolytic protease was estimated to be (38.0±1.5) kDa. As determined by amino acid analysis, it had high contents of asparagine, lysine and serine. The optimum temperature and pH for the collagenase activity were 40 °C and pH=8.0, respectively. The results of the effects of some metal ions, inhibitors and protein substrates suggested that the purified collagenolytic protease is a member of the metalloproteases. Type I collagen (the typical collagen in animal bone) was used as the substrate for determination of Michaelis-Menten kinetics. The obtained Km value was (1.31±0.05) g/L and the corresponding vmax value was (12.54±2.5) μmol/min. The study assumes that the collagenolytic protease purified from B. cereus MBL13 strain could be applied in the hydrolysis of waste animal bones.Izolirana je nova bakterija koja proizvodi kolagenazu, identificirana kao Bacillus cereus MBL13. Iz supernatanta kulture B. cereus MBL13, uzgojene na koštanom kolagenu kao izvoru ugljika i dušika, izdvojena je ekstracelularna proteaza sa sposobnošću razgradnje životinjskih kostiju. Procijenjena je molekularna masa pročišćene kolagenolitičke proteaze od (38,0±1,5) kDa. Analiziran je aminokiselinski sastav i utvrđeno da enzim sadržava veliku količinu asparagina, lizina i serina. Optimalna temperatura za aktivnost kolagenaze bila je 40 °C, a optimalna pH-vrijednost 8,0. Prema utjecaju iona metala, inhibitora i proteinskih supstrata na aktivnost enzima zaključeno je da enzim pripada u grupu metaloproteaza. Kolagen tipa I (tipični sastojak životinjskih kostiju) upotrijebljen je kao supstrat pri određivanju parametara Michaelis-Mentenove kinetike. Km vrijednost bila je (1,31±0,05) g/L, a vmax (12,54±2,5) μmol/min. Zaključeno je da se kolagenolitička proteaza izdvojena iz B. cereus MBL13 može upotrijebiti za razgradnju životinjskih kostiju

Purification and Properties of a Collagenolytic Protease Produced by Bacillus cereus MBL13 Strain

A novel collagenase-producing bacterium has been isolated and identified as Bacillus cereus MBL13. From the culture supernatant of B. cereus MBL13 grown on bone collagen as the sole carbon and nitrogen source, an extracellular protease with novel property of hydrolyzing waste animal bones was purified. The molecular mass of the purified collagenolytic protease was estimated to be (38.0±1.5) kDa. As determined by amino acid analysis, it had high contents of asparagine, lysine and serine. The optimum temperature and pH for the collagenase activity were 40 °C and pH=8.0, respectively. The results of the effects of some metal ions, inhibitors and protein substrates suggested that the purified collagenolytic protease is a member of the metalloproteases. Type I collagen (the typical collagen in animal bone) was used as the substrate for determination of Michaelis-Menten kinetics. The obtained Km value was (1.31±0.05) g/L and the corresponding vmax value was (12.54±2.5) μmol/min. The study assumes that the collagenolytic protease purified from B. cereus MBL13 strain could be applied in the hydrolysis of waste animal bones

Interpretable Hybrid Experiment Learning-Based Simulation Analysis of Power System Planning under the Spot Market Environment

The electricity spot market plays a significant role in promoting the self-improvement of the overall resource utilization efficiency of the power system and advancing energy conservation and emission reduction. This paper analyzes and compares the potential impacts of spot market operations on system planning, considering the differences between planning methods in traditional and spot market environments through theoretical analysis and model comparison. Furthermore, we conduct research and analysis on grid planning methods under the spot market environment with the goal of maximizing social benefits. Unlike the pricing approach based on historical price data in traditional market simulation processes, a data-driven approach that combines experimental economics and machine learning is proposed, specifically using mixed empirical learning to simulate unit bidding strategies in market transactions. A simulation model for electricity spot market trading is constructed to analyze the performance of the planning results in the spot market environment. The case study results indicate that the proposed planning methods can enable the grid to operate well in the spot market environment, maintain relatively stable nodal prices, and ensure the integration of a high proportion of clean energy