The Challenges Facing in Commercial Biotechnology: Its Position in Egypt and Some African Countries

Commercialization of biotechnology can be defined as the conversion of new scientific findings, innovations and discoveries in biotechnology through successful companies and firms or the process by which a product or service in biotechnology is introduced into the general market. Many processes such as sales, production, distribution, marketing, and customer support are required to achieve commercial success. This article deals with factors affecting commercialization of biotechnology in competitive countries and position of biotechnology commercialization in many African countries, including Egypt.

Screening and Media Optimization for Enhancing L-asparaginase Production, an Anticancer Agent, from Different Filamentous Fungi in Solid State Fermentation.

Aim: The aim of present study was to screen new potent fungal isolates and microorganisms possessing extracellular L-asparaginase production capacity. In addition, optimization of cultural and environmental conditions required for enzyme production will be carried out for the highest Lasparaginase producer in solid state fermentation (SSF) technique using agro-industrial residues. Study Design: Screening and physiological studies on the formation of L-asparaginase by Trichoderma viride F2 in order to obtain the optimum cultural and environmental conditions required for enzyme production. Place and Duration of Study: Department of Microbial Chemistry, Genetic Engineering and Biotechnology Division, National Research Centre (NRC), Cairo, Egypt, between July 2013 and June 2015. Methodology: Optimization of physical and nutritional parameters for enzyme production was investigated. Various locally available agro-industrial residues have been screened individually or as mixtures for L-asparaginase production. The combination of Rice husk (RH) with wheat bran (WB) (3:2) proved to be an efficient mixture for enzyme production as it gave the highest enzyme activity (71.87±3.19 U/g-ds) when compared to individual RH (66.71±2.76 U/g-ds) or WB (62.28±2.13 U/g-ds) substrates. Results: Maximal L-asparaginase production (113.43±5.11 U/g-ds) by T. viride F2 was obtained with moisture content of 75%, an inoculums size of 1 x 108 spores/ml and an initial medium pH of 5.0 when incubated at 28ºC for four days. Presence of Tween 20 enhanced enzyme production by 1.19 folds. Glucose (1.0%), Casein (1.5%) and MgCl2 (0.05%) were found to be the best carbon, organic nitrogen and ion sources, respectively. Supplementation of the medium with NaNO3 (0.15%) as an inorganic nitrogen source further increased L-asparaginase production. Under these optimized conditions, L-asparaginase production by T. viride F2 was maximum with a yield of 276.5±13.4 U/g-ds in SSF, which was more than 19-fold enhancement in enzyme activity as compared to that obtained in the basal medium (SmF) (14.23±0.87 U/ml). Conclusion: The results suggest that choosing a suitable substrate coupled with optimization of different parameters can improves enzyme production markedly. Moreover, the production of Lasparaginase from a process based on RH and WB as substrates in SSF is economically attractive due to abundant substrates availability in agriculture-based countries with cheaper cost.

Purification, Kinetic Properties and Antitumor Activity of L-Glutaminase from Penicillium brevicompactum NRC 829.

Aim: The aims of the present study were to purify and characterize L-glutaminase from Penicillium brevicompactum NRC 829; and to evaluate the antitumor activity of the purified enzyme against different tumor human cell lines. Study Design: Testing of antitumor activity of L-glutaminase, purified from a filamentous fungal strain, against four different tumor human cell lines. Place and Duration of Study: Department of Microbial Chemistry, Genetic Engineering and Biotechnology Division, National Research Centre (NRC), Cairo, Egypt, between January 2011 and February 2012. Methodology: P. brevicompactum NRC 829 was grown and maintained on modified Czapek Dox agar (MCD) medium. Cell-free extract was directly used as the source of crude enzyme. L-glutaminase was purified by heat treatment for 20 min at 50ºC, followed by gel filtration on Sephadex G-100 and G-200 columns. Results: An intracellular L-glutaminase from Penicillium brevicompactum NRC 829 was purified to homogeneity (162.75 fold) with an apparent molecular mass (Mr) of 71 kDa. The purified enzyme showed its maximal activity against L-glutamine when incubated at pH 8.5 at 50ºC for 30 min. The purified enzyme retained about 92 % of its initial activity after incubation at 70ºC for 30 min indicating the thermo-stability nature of this enzyme. The highest activity was reported towards its natural substrate, L-glutamine, with an apparent Km value of 1.66 mM. The purified enzyme inhibited the growth of human cell line hepatocellular carcinoma (Hep-G2), with IC50 value of 63.3μg/ml. Conclusion: L-glutaminase purified from Penicillium brevicompactum NRC 829 is a potential candidate in food and pharmaceutical industries.

Purification, Characterization and Antitumor Activity of L-asparaginase from Penicillium brevicompactum NRC 829.

Aim: The aims of this study were to attempt to extract, purify and characterize of Lasparaginase, an antitumor agent, from Penicillium brevicompactum NRC 829. Study Design: Testing of antitumor activity of L-asparaginase against four different tumor human cell lines. Place and Duration of Study: Department of Microbial Chemistry, Genetic Engineering and Biotechnology Division, National Research Centre (NRC), Cairo, Egypt, between June 2010 and November 2011. Methodology: Penicillium brevicompactum NRC 829, a local isolated strain from Culture Collection of the National Research Centre of Egypt, was grown and maintained on modified Czapek Dox medium. The fresh fungal biomass was thoroughly ground with washed cold sand. The cell contents were extracted with cold 0.1M Tris-HCl pH 8.0, thereafter, the slurry obtained was centrifuged at 5500 rpm for 15 min and the supernatant was directly used as the source of enzyme. The purification of L-asparaginase from crudeenzyme extracts of P. brevicompactum was achieved by a sequential multi-steps process starting by heat treatment for 20 min at 50ºC, followed by gel filtration on Sephadex G-100 column, and the most active fractions of L-asparaginase were dialyzed out, lyophilized and then loaded on a Sephadex G-200 column. Results: An intracellular glutaminase-free-L-asparaginase from Penicillium brevicompactum NRC 829 was purified to homogeneity with an apparent molecular mass (Mr) of 94 kDa. The purified enzyme was 151.12 fold with a final specific activity of 574.24 IU/mg protein and about 40% yield recovery. The purified L-asparaginase showed its maximal activity against L-asparagine when incubated at pH 8.0 at 37ºC for 30 min. The enzyme was more stable at alkaline pH than the acidic one and thermally stable up to 60 min at 50-60ºC. L-asparaginase was highly specific for its natural substrate, L-asparagine with a Km value of 1.05 mM. The activity of L-asparaginase is activated by mono cations and various effectors including K+, Na+, 2-mercaptoethanol (2-ME), and reduced glutathione (r-GSH), whereas it is moderately inhibited by various divalent ions including Hg2+, Cu2+, and Ag+. Results indicated the involvement of sulfhydryl group(s) in the enzyme active site(s). The purified enzyme inhibited the growth of human cell line hepatocellular carcinoma (Hep-G2), with IC50 value of 43.3μg/ml. Conclusion: L-asparaginase purified from Penicillium brevicompactum NRC 829 is a potential candidate for medical applications.