Effect of Agitation and Aeration on Keratinase Production in Bioreactors Using Bioprocess Engineering Aspects.

Streptomyces sp. 2M21 was evaluated for keratinase production in bioreactors using chicken feathers. Firstly, optimization of bioengineering parameters (agitation and aeration rates) using Response Surface Methodology was carried out in 2 L bioreactors. Optimized conditions identified by the modified quadratic model were verified as 150 rpm and 1 vvm experimentally corresponding to 351 U/ml of keratinase activity. Moreover, scaling up sequentially to 20 L bioreactors was implemented using constant impeller tip speed and constant mass transfer coefficient as key scale-up parameters. The keratinase activity in 5, 10 and 20 L bioreactors showed similar results with the one of shake flasks (412 U/ml) and 2 L bioreactors (351 U/ml)with respect to the keratinase activity values of 336, 385 and 344 U/ml, respectively. The results suggest keratinase production by evaluating chicken feathers in commercial level. Furthermore, this study has potential to contribute industrial scale production of keratinase by Streptomyces sp. 2M21 using the proposed bioreactor conditions.

Novel keratin modified bacterial cellulose nanocomposite production and characterization for skin tissue engineering.

As it is known that bacterial cellulose (BC) is a biocompatible and natural biopolymer due to which it has a large set of biomedical applications. But still it lacks some desired properties, which limits its uses in many other applications. Therefore, the properties of BC need to be boosted up to an acceptable level. Here in this study for the first time, a new natural nanocomposite was produced by the incorporating keratin (isolated from human hair) to the BC (produced by Acetobacter xylinum) to enhance dermal fibroblast cells' attachment. Two different approaches were used in BC based nanocomposite production: in situ and post modifications. BC/keratin nanocomposites were characterized using SEM, FTIR, EDX, XRD, DSC and XPS analyses. Both production methods have yielded successful results for production of BC based nanocomposite-containing keratin. In vitro cell culture experiments performed with human skin keratinocytes and human skin fibroblast cells indicate the potential of the novel BC/keratin nanocomposites for use in skin tissue engineering.

Optimization of bacterial cellulose production by Gluconacetobacter xylinus using carob and haricot bean.

Bacterial cellulose (BC) can be used in medical, biomedical, electronic, food, and paper industries because of its unique properties distinguishing it from plant cellulose. BC production was statistically optimized by Gluconacetobacter xylinus strain using carob and haricot bean (CHb) medium. Eight parameters were evaluated by Plackett-Burman Design and significant three parameters were optimized by Central Composite Design. Optimal conditions for production of BC in static culture were found as: 2.5g/L carbon source, 2.75g/L protein source, 9.3% inoculum ratio, 1.15g/L citric acid, 2.7g/L Na2HPO4, 30°C incubation temperature, 5.5 initial pH, and 9days of incubation. This study reveals that BC production can be carried out using carob and haricot bean extracts as carbon and nitrogen sources, and CHb medium has higher buffering capacity compared to Hestrin and Schramm media. Model obtained from this study is used to predict and optimize BC production yield using CHb medium.

Microbial ribonucleases (RNases): production and application potential.

Ribonuclease (RNase) is hydrolytic enzyme that catalyzes the cleavage of phosphodiester bonds in RNA. RNases play an important role in the metabolism of cellular RNAs, such as mRNA and rRNA or tRNA maturation. Besides their cellular roles, RNases possess biological activity, cell stimulating properties, cytotoxicity and genotoxicity. Cytotoxic effect of particular microbial RNases was comparable to that of animal derived counterparts. In this respect, microbial RNases have a therapeutic potential as anti-tumor drugs. The significant development of DNA vaccines and the progress of gene therapy trials increased the need for RNases in downstream processes. In addition, RNases are used in different fields, such as food industry for single cell protein preparations, and in some molecular biological studies for the synthesis of specific nucleotides, identifying RNA metabolism and the relationship between protein structure and function. In some cases, the use of bovine or other animal-derived RNases have increased the difficulties due to the safety and regulatory issues. Microbial RNases have promising potential mainly for pharmaceutical purposes as well as downstream processing. Therefore, an effort has been given to determination of optimum fermentation conditions to maximize RNase production from different bacterial and fungal producers. Also immobilization or strain development experiments have been carried out.