Journal of Applied Sciences Research, 3(10): 921-925, 2007 © 2007, INSInet Publication Corresponding Author: I.S. Abd-Elsalam, Department of Natural and Microbial products chem. 921 Factorial Design for the Production of Some Steroidal Hormones from Sunflower Oil Sterols by Fusarium solani I.S. Abd-Elsalam, H.A. El-Refai and A.M. Abouel-Fotouh 1 1 2 Department of Natural and Microbial products chem. 1 Department of Mech. Eng. (National research center) 2 Abstract: The bioconversion of plant sterols (phytosterols) by Fusarium solani to produce some C-19 androstene steroids namely androstenedione (AD) and androstadienedione (ADD) has been investigated. The purpose of this study was to determine the conditions for maximum yields of both AD and ADD using factorial design and multi-dimensional response surface (RSM) modeling techniques. Therefore, the effect of three factors (pH, biotransformation time and substrate level on the yields of both AD and ADD were assessed using a sequential factorial design. The results indicated that the biotransformation time was critical for the production of AD and ADD, while the pH value of the fermentation medium affected the yield of both AD and ADD to a lesser extent. An optimization experiment was performed using factor levels within narrower range of the setting determined from the first experiment, the yield of AD and ADD predicted by the mathematical model increased to 65.71 and 20.41 %, respectively and this value were confirmed experimentally. Key words: Bioconversion, Sunflower sterols, Factorial design, Androstene, Steroid. INTRODUCTION Although the steroid drugs represent only a small part of the world market of pharmaceuticals, there is a great demand for new and cheaper steroid raw materials for their production . The choice of starting materials has [2,5,3,4] always had a critical, impact on steroid manufacturing industries. The most common and economical process, for the production of steroid pharmaceuticals is the partial synthesis from relatively inexpensive steroid row materials of animal and plant origins. One of the most important bioconversion reaction of steroid is the side chain degradation of sterols specially, cholesterol. b-sitosterol,which had been studied . The [7,12] biodegredation products were the highly active steroidal hormones AD and ADD, which are important intermediates in the production of estrone and estradiol. There are different strains which have the ability to degrade phytosterols. The highly active strains were Fusarium spp, Aspergillus spp. in addition to some bacterial strains. The studies of El- Refai and Abd- Elsalam , stated that the strain of F. solni has the ability [6] to transform â- sitosterol, as well as sun flower sterols mixture to gave a good yields of both AD, ADD. The work was extended in this communication to build a mathematical design for optimizing the bioconversion 19 process in favour of the desired C androgenes. MATERIALS AND METHODS Materials: Microorganisms: Fusarium solani was kindly provided from the Natural and Microbial Products Chemistry Department, National Research Centre (NRC) Dokki, Cairo, Egypt. Chemicals: The authentic steroids used (AD, ADD, â-sitosterol), and 8-hydroxyquinoline were provided by of Sigma Company USA. Sunflower oil used in the current work is of commercial grade. Methods: Maintenance of the Microorganism: The experimental organism was maintained on the following medium g/l (malt extract 25, yeast extract 4 and agar 20) and was monthly interval regenerated. Transformation process: Cultivation was performed using 250 ml Erlenmeyer flasks, each containing 50 ml of 4 2 4 the following medium (g/l): glucose, 10; (NH ) SO , 1.0, 2 4 2 4 K HPO , 7.0; KH P0 ,3.0. To each flask 0.016 g/1 8- hydroxyquinoline was added to avoid the enzymatic cission of the sterol nucleus . The pH was adjusted to [8] 6.5. The flasks were sterilized using steam autoclaving at 120 C for 20 min and incubated with 2 ml spore o suspension of F. solani 7 days old culture. The culture flasks were agitated on reciprocal shaker (200 rpm) at 30°C±for 72h . Thereafter, the sunflower sterols mixture (5 mg) was added to each flask and the transformation process was continued for the specific time intervals . [6] Extraction: At the end of the transformation period, the content of each flask was homogenized in a blender (16000 rpm) with double its volume of chloroform. The chloroform layer was separated and dried under vacuum to give semi-solid residue "test material".