RESUMO
Novel Geobacillus sp. DS3, isolated from the Sikidang Crater in Dieng, exhibits promising characteristics for industrial applications, particularly in thermostable α-amylase production. Recombinant technology was used to express thermostable α-amylase in E. coli BL21(DE3) to overcome high-temperature production challenges. The study aimed to express, purify, characterize, and explore potential applications of this novel enzyme. The enzyme was successfully expressed in E. coli BL21(DE3) at 18 °C for 20 h with 0.5 mM IPTG induction. Purification with Ni-NTA column yielded 69.23 % from the initial crude enzyme, with a 3.6-fold increase in specific activity. The enzyme has a molecular weight of ±70 kDa (±58 kDa enzyme+11 kDa SUMO protein). It exhibited activity over a wide temperature range (30-90 °C) and pH range (6-8), with optimal activity at 70 °C and pH 6 with great stability at 60 °C. Kinetic analysis revealed Km and Vmax values of 324.03 mg/ml and 36.5 U/mg, respectively, with dextrin as the preferred substrate without cofactor addition. As a metalloenzyme, it showed the best activity in the presence of Ca2+. The enzyme was used for porous starch production and successfully immobilized with chitosan, exhibiting improved thermal stability. After the fourth reuse, the immobilized enzyme maintained 62 % activity compared to the initial immobilization.
RESUMO
Incorporating antimicrobial components into food packaging materials can prevent microbial contamination. Fungus combs could be an alternative source of natural antimicrobial agents. In this study, n-hexane, ethyl acetate, methanol, and water extracts were obtained from fungus combs isolated from Indomalayan termite (Macrotermes gilvus Hagen) mound. Their antibacterial and antifungal activities against food spoilage microorganisms including Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus ATCC 25923, Aspergillus flavus, and Aspergillus niger were evaluated by Kirby-Bauer disc diffusion and microdilution. Results showed that ethyl acetate extract formed the largest diameter inhibition zone for all tested bacteria and fungi, exhibited antibacterial activity against all tested bacteria with minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of 0.39 and 0.78 mg/mL, respectively, and suppressed A. flavus and A. niger with an MIC value of 0.78 mg/mL. This extract contained guaiacol and syringol, which were predicted as the main antimicrobial components in fungus comb. n-Hexane extract only inhibited Gram-positive bacteria. S. aureus ATCC 25923 was the most sensitive to all the extracts, and A. flavus was more sensitive than A. niger. All these fungus comb extracts exhibited antimicrobial activity against E. coli ATCC 25922, P. aeruginosa ATCC 27853, S. aureus ATCC 25923, A. flavus, and A. niger. This study revealed that fungus comb extracts, especially ethyl acetate, could be considered as a new antimicrobial agent.