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Genotyping of HCV in Egyptians & response to treatment
S460
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
Acknowledgement The authors thank the Centre of Analysis and Test of Huazhong University of Science and Technology for LC-MSn analysis. doi:10.1016/j.jbiotec.2010.09.677 [P-M.90] Genotyping of HCV in Egyptians & response to treatment S. Nasr 1,∗ , C. Trepo 2 , J. Grimaud 3 , H. El-Boushara 4 1
NCRRT-Cytopathology&Immunology -Egypt AT-Eng, Egypt INSERM-U271-Lyon, France 3 Universite Paris-VI, France 4 Homs University, Syrian Arab Republic Keywords: HCV; Genotyping; HCV-treatment; HCC 2
Hepatitis C virus (HCV) is a major cause of acute hepatitis and chronic liver disease, including cirrhosis and liver cancer.Globally,an estinated 170 million persons are chronically infected with HCV and 3 out of 4 million persons are infected each year. In view of the absence of vaccination, and the treatment is very costly. doi:10.1016/j.jbiotec.2010.09.678 [P-M.91] Strain improvement of Curvularia lunata for production of hydrocortisone V.V. Kollerov, A.A. Shutov, V.V. Fokina, G.V. Sukhodolskaya, S.M. Khomutov, M.V. Donova ∗ IBPM RAS, Russian Federation Keywords: Steroid; Bioconversion; Curvularia lunata; Hydrocortisone Physiologically active 11-hydroxylated steroids are widely used in medicine as anti-inflammatory and anti-allergic agents (Fernandes et al., 2003). The insertion of hydroxyl group into 11-position of steroidal molecule could be realized by the use of filamentous fungi. The maximal 11-hydroxylase activity was observed for Curvularia lunata (Cochliobolus lunatus) strains. There are many approaches to increase the yield of 11hydroxylated products during cortexolone transformation with C. lunata (Sukhodolskaya et al., 1993; Kelly et al., 2003) However, low selectivity and not high productivity of the processes using the known strains remain the “bottleneck” at the further practical application of the biotechnological methods developed. The present study was purposed to obtain mutant strains of C. lunata with increased 11-hydroxylase activity towards cortexolone and its derivatives: 17␣-acetate, 21-acetate and 17␣,21-diacetate and to work out the biotechnological method for the effective production of hydrocortisone and its acetylated derivatives. The original procedures of protoplasts formation, mutagenesis and selection of the mutants have been worked out. The mutant strain of C. lunata M4 with high 11-hydroxylase activity towards 21-acetate and 17␣,21-diacetate of cortexolone was obtained. It was shown that the presence of 17␣-acetate group facilitates 11-hydroxylation with the repression of undesirable 14␣-hydroxylation of acetylated steroid substrate. The yield of sum of 11-hydroxylated products: hydrocortisone and its 17␣-acetate
Fig. 1. Transformation of 17␣,21-diacetate of cortexolone (1 g/l) with the original and mutant M4 strains of C. lunata.
reached 90% at the transformation of cortexolone 17␣,21-diacetate (1 g/l) using mutant M4 (Fig. 1). Application of the original procedures of substrate micronization and addition to the fermentation medium provided high yields of the 11-hydroxylated derivatives (over 75%) even at 34-fold increase of the substrate loadings. The process was carried out at the laboratory fermenter (3L). The effective procedure of crystalline hydrocortisone recovery provided obtaining of pharmacopoeia quality product. The results evidenced the advantages of mutant strain M4 application for biotechnological production of hydrocortisone from 17␣,21-diacetate of cortexolone. References Fernandes, P., Cruz, A., Angelova, B., Pinheiro, H.M., 2003. Microbial conversion of steroid compounds. recent developments. Ensyme and Microbial Technology 32, 688–705. G. V. Sukhodolskaya, B. Angelova, K. A. Koscheenko, I. M. Basovskaya, G. K Skryabin. 1993, RF Patent, 1411336. Kelly, S.L., Lamb, D.C., Jackson, C.J., Warrilow, A.G., Kelly, D.E., 2003. The biodiversity of microbial cytochromes P450. Adv. Microb. Physiol 47, 131–186.
doi:10.1016/j.jbiotec.2010.09.679 [P-M.92] Genotyping of HCV-Egyptian infected cases, and response to treatment S. Nasr 1,2,3,∗ , C. Trepo 1,2,3 , J. Grimaud 1,2,3 , H. El-Booushara 1,2,3 1
NCRRT-AT-Eng, Egypt INSERM-U271-Lyon, France 3 University Paris 6, France4 Homs university, Syrian Arab Republic Keywords: HCV infection; HCC; Genotyping & treatment; Interferon alfa 2
Hepatitis C virus (HCV) is a major cause of acute hepatitis and chronic liver disease, including cirrhosis and liver cancer. Globally, an estimated 170 million persons are chronically infected with HCV and 3 out of 4 million persons are infected each year. In view of the absence of vaccination, and the treatment is too costly for most countries in developing countries to afford .from
Special Abstracts / Journal of Biotechnology 150S (2010) S1–S576
Acknowledgement The authors thank the Centre of Analysis and Test of Huazhong University of Science and Technology for LC-MSn analysis. doi:10.1016/j.jbiotec.2010.09.677 [P-M.90] Genotyping of HCV in Egyptians & response to treatment S. Nasr 1,∗ , C. Trepo 2 , J. Grimaud 3 , H. El-Boushara 4 1
NCRRT-Cytopathology&Immunology -Egypt AT-Eng, Egypt INSERM-U271-Lyon, France 3 Universite Paris-VI, France 4 Homs University, Syrian Arab Republic Keywords: HCV; Genotyping; HCV-treatment; HCC 2
Hepatitis C virus (HCV) is a major cause of acute hepatitis and chronic liver disease, including cirrhosis and liver cancer.Globally,an estinated 170 million persons are chronically infected with HCV and 3 out of 4 million persons are infected each year. In view of the absence of vaccination, and the treatment is very costly. doi:10.1016/j.jbiotec.2010.09.678 [P-M.91] Strain improvement of Curvularia lunata for production of hydrocortisone V.V. Kollerov, A.A. Shutov, V.V. Fokina, G.V. Sukhodolskaya, S.M. Khomutov, M.V. Donova ∗ IBPM RAS, Russian Federation Keywords: Steroid; Bioconversion; Curvularia lunata; Hydrocortisone Physiologically active 11-hydroxylated steroids are widely used in medicine as anti-inflammatory and anti-allergic agents (Fernandes et al., 2003). The insertion of hydroxyl group into 11-position of steroidal molecule could be realized by the use of filamentous fungi. The maximal 11-hydroxylase activity was observed for Curvularia lunata (Cochliobolus lunatus) strains. There are many approaches to increase the yield of 11hydroxylated products during cortexolone transformation with C. lunata (Sukhodolskaya et al., 1993; Kelly et al., 2003) However, low selectivity and not high productivity of the processes using the known strains remain the “bottleneck” at the further practical application of the biotechnological methods developed. The present study was purposed to obtain mutant strains of C. lunata with increased 11-hydroxylase activity towards cortexolone and its derivatives: 17␣-acetate, 21-acetate and 17␣,21-diacetate and to work out the biotechnological method for the effective production of hydrocortisone and its acetylated derivatives. The original procedures of protoplasts formation, mutagenesis and selection of the mutants have been worked out. The mutant strain of C. lunata M4 with high 11-hydroxylase activity towards 21-acetate and 17␣,21-diacetate of cortexolone was obtained. It was shown that the presence of 17␣-acetate group facilitates 11-hydroxylation with the repression of undesirable 14␣-hydroxylation of acetylated steroid substrate. The yield of sum of 11-hydroxylated products: hydrocortisone and its 17␣-acetate
Fig. 1. Transformation of 17␣,21-diacetate of cortexolone (1 g/l) with the original and mutant M4 strains of C. lunata.
reached 90% at the transformation of cortexolone 17␣,21-diacetate (1 g/l) using mutant M4 (Fig. 1). Application of the original procedures of substrate micronization and addition to the fermentation medium provided high yields of the 11-hydroxylated derivatives (over 75%) even at 34-fold increase of the substrate loadings. The process was carried out at the laboratory fermenter (3L). The effective procedure of crystalline hydrocortisone recovery provided obtaining of pharmacopoeia quality product. The results evidenced the advantages of mutant strain M4 application for biotechnological production of hydrocortisone from 17␣,21-diacetate of cortexolone. References Fernandes, P., Cruz, A., Angelova, B., Pinheiro, H.M., 2003. Microbial conversion of steroid compounds. recent developments. Ensyme and Microbial Technology 32, 688–705. G. V. Sukhodolskaya, B. Angelova, K. A. Koscheenko, I. M. Basovskaya, G. K Skryabin. 1993, RF Patent, 1411336. Kelly, S.L., Lamb, D.C., Jackson, C.J., Warrilow, A.G., Kelly, D.E., 2003. The biodiversity of microbial cytochromes P450. Adv. Microb. Physiol 47, 131–186.
doi:10.1016/j.jbiotec.2010.09.679 [P-M.92] Genotyping of HCV-Egyptian infected cases, and response to treatment S. Nasr 1,2,3,∗ , C. Trepo 1,2,3 , J. Grimaud 1,2,3 , H. El-Booushara 1,2,3 1
NCRRT-AT-Eng, Egypt INSERM-U271-Lyon, France 3 University Paris 6, France4 Homs university, Syrian Arab Republic Keywords: HCV infection; HCC; Genotyping & treatment; Interferon alfa 2
Hepatitis C virus (HCV) is a major cause of acute hepatitis and chronic liver disease, including cirrhosis and liver cancer. Globally, an estimated 170 million persons are chronically infected with HCV and 3 out of 4 million persons are infected each year. In view of the absence of vaccination, and the treatment is too costly for most countries in developing countries to afford .from