Background A wide variety of bacterial species produces protease enzyme, and the application of the same enzyme has been manipulated precisely and used in various biotechnological areas including industrial and environmental sectors. The main aim of this research study was to isolate, screen, and identify alkaline protease-producing bacteria that were sampled from leather industry effluent present in the outer skirts of Addis Ababa, Ethiopia. Purpose To isolate and characterize the alkaline protease-producing bacteria from leather industrial effluents. Methods Samples are collected from Modji leather industrial effluents and stored in the microbiology lab. After isolated bacteria from effluent using serial dilution and followed by isolated protease-producing bacteria using skim milk agar media. After studying primary and secondary screening using zonal inhibition methods to select potential protease-producing bacteria using skim milk agar media. Finally, to identify the potential bacteria using biochemical methods, bacterial biomass, protease activity, and gene sequencing (16S rRNA) method to finalize the best alkaline protease producing bacteria identified. Results First twenty-eight different bacterial colonies were isolated initially from the leather industry effluent sample situated at the Modjo town of Ethiopia. The isolated bacteria were screened using the primary and secondary screening method with skim milk agar medium. At the primary level, we selected three isolates namely ML5(14 mm), ML12(18 mm), and MS12 (15 mm), showing the highest zone of proteolysis as a result of casein degradation on the agar plates were selected and subjected to primary screening. Further secondary screening confirmed that the zone of inhibition methods ML5 (14.00±0.75 mm), ML12 (19.50±0.66 mm), and MS12 (15.00±1.32 mm) has efficient proteolytic activity and can be considered as effective protease producer. The three isolates were then subjected to morphological and biochemical tests to identify probably bacterial species, and all the three bacterial isolates were found out to be of Bacillus species. The shake flask method was carried out to identify the most potent one having greater biomass production capabilities and protease activity. ML12 isolated from leather effluent waste showed the highest protease activity (19 U/ml), high biomass production, and the same was subjected to molecular identification using 16s sequencing and a phylogenetic tree was constructed to identify the closest neighbor. The isolate ML12 (Bacillus cereus strain -MN629232.1) is 97.87% homologous to Bacillus cereus strain (KY995152.1) and 97.86% homologous to Bacillus cereus strain (MK968813.1). Conclusions This study has exposed that from twenty-eight different bacterial samples isolated from leather industry effluent; further primary and secondary screening methods were selected three potential alkaline protease strains. Finally, based on its biochemical identification, biomass, and protease activity, ML12 (Bacillus cereus strains) is the best strain identified. The alkaline protease has the significant feature of housing potent bacterial species for producing protease of commercial value.

In recent years, there has been an increase in demand for bioactive techniques in the food packaging industry. Although edible packaging is popular, it has yet to be effectively implemented into the market. Packaging made of plastics and chemicals is widely employed in the market today, posing a threat to the environment and living creatures. This research attempts to show current breakthroughs and progress in the field of biodegradable packaging. When compared to ancient packaging materials, bio-based packaging materials are safer. Sustainable biodegradable packaging materials can be made from edible films, coatings, and other bio food packaging techniques made from various biological resources. This paper discusses the important qualities and advantages of several bio-based packing materials. It is highlighted the advantages of bio-based packaging materials over synthetic packaging materials. It has been debated the importance of employing bio-based packaging to mitigate the environmental risks associated with traditional packaging technologies. Many researchers may be prompted by this study to focus on packaging reformulation options. Thus, we can attain food packing materials by considering customer's economic and sustainability aspects.

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Protease is an enzyme which has a wide range of applications in various fields. Extracellular protease was produced from Pseudomonas aeruginosa and Enterococcus hirae which were isolated from the effluents of diary industries. Protease immobilized with super paramagnetic nanoparticles was characterized by DLS, XRD and TEM methods in relation to their size and structure. The protease enzyme was bound to magnetic nanoparticles via surface transformation technique including Silica coated magnetic Nano composite, amine and cysteine functioned Nano composite formation. Successful binding of protease onto the particles was confirmed by TEM imaging. The maximal enzyme activity of immobilized protease was determined using universal protease assay and was found to be 105µg mL -1 & 290µg mL -1 for Pseudomonas sp. and Enterococcus sp. respectively. The immobilization capacity of protease onto nanoparticles was 6000µM/g. The stability of the immobilized enzyme increased in comparison with the free enzyme. Overall, this study showed that the stability and activity of the protease was enhanced by immobilization to the magnetic nanoparticles. This suggested that immobilized enzyme on the magnetic beads of nanoparticles could be used in an interesting range of applications, both in broader temperature and pH ranges, also permitting magnetic recovery of the enzyme for reuse or purification of the product.

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