In vitro molecular study of wound healing using biosynthesized bacteria nanocelluse/silver nanocomposite assisted by bioinformatics databases

Background: In recent years, bacterial nanocellulose (BNC) based nanocomposites have been developed to promote healing property and antibacterial activity of BNC wound dressing. Molecular study can help to better understanding about interaction of genes and pathways involved in healing progression. Objectives: The aim of this study was to prepare bacterial nanocellulose/silver (BNC/Ag) nanocomposite films as ecofriendly wound dressing in order to assess their physical, cytotoxicity and antimicrobial properties. The in vitro molecular study was performed to evaluate expression of genes involved in healing of wounds after treatment with BNC/Ag biofilms. Study design, materials, and methods: Silver nanoparticles were formed by using Citrullus colocynthis extract within new isolated bacterial nanocellulose (BNC) RM1. The nanocomposites were characterized using X-ray diffraction, Fourier transform infrared, and field emission scanning electron microscopy. Besides, swelling property and Ag release profile of the nanocomposites were studied. The ability of nanocomposites to promote wound healing of human dermal fibroblast cells in vitro was studied. Bioinformatics databases were used to identify genes with important healing effect. Key genes which interfered with healing were studied by quantitative real time PCR. Results: Spherical silver nanoparticles with particle size ranging from 20 to 50 nm were synthesized and impregnated within the structure of BNC. The resulting nanocomposites showed significant antibacterial activities with inhibition zones ranging from 7±0.25 to 16.24±0.09 mm against skin pathogenic bacteria. Moreover, it was compatible with human fibroblast cells (HDF) and could promote in vitro wound healing after 48h. Based on bioinformatics databases, the genes of TGF-β1, MMP2, MMP9, CTNNB1, Wnt4, hsa-miR-29b-3p and hsa-miR-29c-3p played important role in wound healing. The nanocomposites had an effect in expression of the genes in healing. Thus, the BNC/Ag nanocomposite can be used to heal wound in a short period and simple manner. Conclusion: This eco-friendly nanocomposite with excellent antibacterial activities and healing property confirming its utility as potential wound dressings

Pullulanase type II from a local Bacillus isolate H1.5 : isolation, production and characterization of the enzyme

Enzymes that specifically break down the branch points – debranching enzymes – have been known for more than a decade. Pullulanase (EC 3.2.1.41, pullulan 6- glucanohydrolase), which was first reported in Aerobacter aerogenes splits the α-1,6- glucosidic linkages in pullulan and give rise finally to maltotriose (Yuzuru and Masaharu, 1983). In most cases, pullulanase can also cleave the α-1,6 link of other branched polysaccharides, such as starch, glycogen and maltodextri

Large scale production of liquid wax ester by immobilized lipase

Oleyl oleate, a liquid wax ester was synthesized by an immobilized Candida antartica lipase B (Novozym 435) as biocatalyst using oleic acid and oleyl alcohol. The effects of various reaction parameters were optimized to obtain a high yield of liquid wax esters. Investigation in large scale production was performed in batch mode of stirred tank reactor (STR) with one multi-bladed impeller. The optimum condition to produce liquid wax ester was, reaction time (RT); 30 min, temperature (T); 50 °C, amount of enzyme (E); 90 g (900,000 PLU), agitation speed (A) of 400 rpm, number of impeller tip (N) of 2 and molar ratio of oleyl alcohol to oleic acid (M); 2:1. Analysis of the yield showed that at optimum condition, >90% liquid wax esters were produced. The stability of Novozym 435 showed at high percentage (>80%) up to 4 cycles

High performance enzymatic synthesis of oleyl oleate using immobilised lipase from Candida antartica

High performance enzymatic synthesis of oleyl oleate, a liquid wax ester was carried out by lipase-catalysed esterification of oleic acid and oleyl alcohol. Various reaction parameters were optimised to obtain high yield of oleyl oleate. The optimum condition to produce oleyl oleate was reaction time; 5 min, organic solvents of log P 65 3.5, temperature; 40-50\ub0C, amount of enzyme; 0.2-0.4 g and molar ratio of oleyl alcohol to oleic acid; 2:1. The operational stability of enzyme was maintained at >90% yield up to 9 cycles. Analysis of the yield of the product showed that at optimum conditions, >95% liquid wax esters were produced

Biosynthesis of ZnO nanoparticles by a new yeast strain Pichia kudriavzevii and evaluation of their antimicrobial and antioxidant activities

The potential ability of a new yeast strain, Pichia kudriavzevii, in the synthesis of zinc oxide nanoparticles (ZnO-NPs) through a green method was explored in this study. The effect of reaction time (12, 24 and 36 h) on the structure of the resulting ZnO nanoparticles was investigated. From the XRD and TEM results, the ZnO-NPs with a hexagonal wurtzite structure and a particle crystal size of ~10-61 nm was formed at different reaction times. Combing XRD, TEM, and PL results, it was revealed that the sample prepared at intermediate duration (24 h) has the most favorable nanosized structure with the lowest defect concentration. The biomedical properties of ZnO-NPs as free radical scavenging activity, cytotoxicity and antibacterial agents were characterized. Biosynthesized ZnO-NPs showed strong DPPH free radical scavenging and a dose dependent toxicity with non-toxic effects on Vero cells for concentrations below 190 µg/mL. Desirable bactericidal activity was shown by the ZnO-NPs on Gram-positive bacteria (Bacillus subtilis, Staphylococcus epidermidis and Staphylococcus aurous) and Gram-negative bacteria (Escherichia coli and Serratia marcescens). A maximum inhibition zone of ~19 mm was observed for Staphylococcus epidermidis at a concentration of 100 µg/mL for sample prepared at 24 h. The results from this study reveal that ZnO-NPs possesses potential for many medical and industrial applications

High performance enzymatic synthesis of oleyl oleate using immobilised lipase from Candida antartica

High performance enzymatic synthesis of oleyl oleate, a liquid wax ester was carried out by lipase-catalysed esterification of oleic acid and oleyl alcohol. Various reaction parameters were optimised to obtain high yield of oleyl oleate. The optimum condition to produce oleyl oleate was reaction time; 5 min, organic solvents of log P ≥ 3.5, temperature; 40-50ºC, amount of enzyme; 0.2-0.4 g and molar ratio of oleyl alcohol to oleic acid; 2:1. The operational stability of enzyme was maintained at >90% yield up to 9 cycles. Analysis of the yield of the product showed that at optimum conditions, >95% liquid wax esters were produced

Evaluation of commercial soy sauce koji strains of Aspergillus oryzae for γ-aminobutyric acid (GABA) production

In this study, four selected commercial strains of Aspergillus oryzae were collected from soy sauce koji. These A. oryzae strains designated as NSK, NSZ, NSJ and NST shared similar morphological characteristics with the reference strain (A. oryzae FRR 1675) which confirmed them as A. oryzae species. They were further evaluated for their ability to produce γ-aminobutyric acid (GABA) by cultivating the spore suspension in a broth medium containing 0.4 % (w/v) of glutamic acid as a substrate for GABA production. The results showed that these strains were capable of producing GABA; however, the concentrations differed significantly (P < 0.05) among themselves. Based on the A. oryzae strains, highest GABA concentration was obtained from NSK (194 mg/L) followed by NSZ (63 mg/L), NSJ (51.53 mg/L) and NST (31.66 mg/L). Therefore, A. oryzae NSK was characterized and the sequence was found to be similar to A. oryzae and A. flavus with 99 % similarity. The evolutionary distance (K nuc) between sequences of identical fungal species was calculated and a phylogenetic tree prepared from the K nuc data showed that the isolate belonged to the A. oryzae species. This finding may allow the development of GABA-rich ingredients using A. oryzae NSK as a starter culture for soy sauce production

Purification of kojic acid

The present invention provides a method of purifying kojic acid from fermentation broth by repeated crystallization in solvents to obtain suitable kojic acid crystal size and structure, which is free from impurities such as pigments, metals and the like. Accordingly, the present method comprises the steps of: (i) obtaining kojic acid solution from a fermentation broth; (ii) dissolving the kojic acid solution in a solvent or a mixture of solvents with appropriate volume to maintain the kojic acid solution at a concentration of 80 g/l to 100 g/l; (iii) agitating the concentrated kojic acid solution at 50 to 100 rpm; (iv) removing the solvent or mixture of solvents contained in the concentrated kojic acid solution by way of evaporation at 50°C to 80°C; (v) maintaining the temperature of the concentrated kojic acid solution at 5°C to 35°C for up to 10 hour to form kojic acid crystals; (vi) separating the kojic acid crystals; and (vii) repeating the steps of (ii) through (vi) for more than once in order to form high purity kojic acid crystals, which are free from impurities