Biochemical characterization of purified phytase produced from Aspergillus awamori AFE1 associated with the gastrointestinal tract of longhorn beetle (Cerambycidae latreille).

The need for industrially and biotechnologically significant enzymes, such as phytase, is expanding daily as a result of the increased use of these enzymes in a variety of operations, including the manufacture of food, animal feed, and poultry feed. This study sought to characterize purified phytase from A. awamori AFE1 isolated from longhorn beetle for its prospect in industrial applications. Ammonium sulfate precipitation, ion-exchange chromatography, and gel-filtration chromatography were used to purify the crude enzyme obtained from submerged fermentation using phytase-producing media, and its physicochemical characteristics were examined. The homogenous 46.8-kDa phytase showed an 8.1-fold purification and 40.7% recovery. At 70 C and pH 7, the optimum phytase activity was noted. At acidic pH 4-6 and alkaline pH 8-10, it likewise demonstrated relative activity of 88-95% and 67-88%, respectively. It showed 67-70% residual activity between 30 and 70 C after 40 min, and 68-94% residual activity between pH 2 and 12 after 2 h. The presence of Hg+, Mg2+, and Al3+ significantly decreased the enzymatic activity, whereas Ca2+ and Cu2+ enhanced it. Ascorbic acid increased the activity of the purified enzyme, whereas ethylenediaminetetraacetic acid (EDTA) and mercaptoethanol inhibited it. The calculated values for Km and Vmax were 55.4 mM and1.99 µmol/min/mL respectively. A. awamori phytase, which was isolated from a new source, showed unique and remarkable qualities that may find use in industrial operations such as feed pelleting and food processing.

Purification and biochemical characterization of pullulanase produced from Bacillus sp. modified by ethyl-methyl sulfonate for improved applications.

Strain improvement via chemical mutagen could impart traits with better enzyme production or improved characteristics. The present study sought to investigate the physicochemical properties of pullulanase produced from the wild Bacillus sp and the mutant. The pullulanases produced from the wild and the mutant Bacillus sp. (obtained via induction with ethyl methyl sulfonate) were purified in a-three step purification procedure and were also characterized. The wild and mutant pullulanases, which have molecular masses of 40 and 43.23 kDa, showed yields of 2.3% with 6.0-fold purification and 2.0% with 5.0-fold purification, respectively, and were most active at 50 and 40 °C and pH 7 and 8, respectively. The highest stability of the wild and mutant was between 40 and 50 °C after 1 h, although the mutant retained greater enzymatic activity between pH 6 and 9 than the wild. The mutant had a decreased Km of 0.03 mM as opposed to the wild type of 1.6 mM. In comparison to the wild, the mutant demonstrated a better capacity for tolerating metal ions and chelating agents. These exceptional characteristics of the mutant pullulanase may have been caused by a single mutation, which could improve its utility in industrial and commercial applications.

Production of herbal toothpaste: Physical, organoleptic, phyto-compound, and antimicrobial properties.

Objective: To investigate the possibility of producing dental antimicrobial toothpaste from Allium cepa L skin chaff, Azadirachta indica A. seed, and Tetrapleura tetraptera pod extracts. Methods: Ethanolic extracts of the three plant materials were obtained. These were subjected to phytochemical and GC-MS analyses. The different extract combinations were used for the production of various toothpaste. The toothpaste's physical, organoleptic, and antimicrobial properties were determined. Results: From the phytochemical analysis, Allium cepa has the highest phenolic (1.20 mgGAE/g), saponin (14.80%), tannin (0.11 mg/g) and DPPH (82.80%), Tetrapleura tetraptera has the highest flavonoid (0.33 mg RE/g), and alkaloid (20.50 mg/g) while, Azadirachta indica has the highest oxalate (77.50 mg/g). The GC-MS revealed significant chemical components of Allium cepa as 1-heptatriacotanol, germacra-1(10),4,11(13)-trien-12-oic acid, 6-alpha-hydroxy-,gamma-lactone, (E,E)-, 11H-Indeno [1,2b] quinoxaline, 2-methyl- while Azadirachta indica have butyl benzoate, benzoic acid, hexyl ester, hexadecanoic acid, methyl ester and Tetrapleura tetraptera have the following 15-hydroxypentadecanoic acid, cis-9-hexadecenal, and 11,13-dimethyl-12-tetradecen-1-ol acetate. All the produced toothpaste has a brown colour and a pleasant smell, with pH from 7.30 to 8.10 and foamability from 19.23% of stand-alone toothpaste to 44.44% of Allium cepa-based. Amongst the produced toothpaste Allium cepa-based toothpaste has the best antimicrobial activities against the tested bacteria (Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca) and fungi (Candida albicans and Candida parapsilosis). The stand-alone toothpaste has the lowest minimum inhibitory concentration of 1.25 mg/mL against bacteria and fungi. Clinical significance: The study provides information on the production of human health-friendly dental antimicrobial toothpaste from plant materials.