|
|
|
| Studies on the 3-Ketosteriod-1-Dehydrogenation of Steroid Hormone by Cellular lysates of Mycobacterium |
| QIN Meng-fei, SUN Hong, SONG Hao |
| School of Chemical Engineering and Technology, Key Laboratory of Systems Bioengineering, Ministry of Education, SynBio Research Platform, Collaborative Innovation Center of Chemical Science and Engineering, Tianjin University, Tianjin 300072, China |
|
|
|
Abstract 9β, 11β-Epoxypregn-4-ene-17α, 21-diol-3, 20-dione 21-acetate (Ⅰ) is a substrate for the production of 9β, 11β-Epoxypregn-1, 4-diene-17α, 21-diol-3, 20-dione (Ⅳ), which is a key precursor for the production of many 9-fluoro-substituted corticosteroid hormones. By comparing whole cells catalysis and cellular lysates conversion, it was found that whole cells of Mycobacterium sp. MS136 could only convertⅠto 9β, 11β-Epoxypregn-4-ene-17α, 21-diol-3, 20-dione (Ⅱ), and Ⅰ can be effectively converted toⅣ by cellular lysates.The reaction order is that Ⅰ is spontaneously hydrolyzed to Ⅱ and Ⅱundergoes C1, 2-dehydrogenation reaction toⅣ.In order to improve the productivity of Ⅳ, the key genes kstD, kstD3 and kstDM encoding C1, 2-dehydrogenase (KSTD)were overexpressed in Mycobacterium sp. MS136 to enhance the C1, 2-dehydrogenation reaction rate, and the results showed that 1 g/L substrate Ⅰ can be converted by recombinant strain MS136-kstDM cellular lysates at pH 7.0, the productivity of Ⅳreached 78.4% after 45 h, which is 38.9% higher than original strain.The reaction rate is enhanced by optimizing the pH, and the results showed that 1 g/L substrate (Ⅰ) can be converted by recombinant strain MS136-kstDM cellular lysates at pH 7.5, the productivity of Ⅳreached 92.8% after 45 h, which was 63.4% higher than original strain.
|
|
Received: 23 February 2017
Published: 25 August 2017
|
|
|
|
|
[1] Tong W Y, Dong X. Microbial biotransformation:recent developments on steroid drugs. Recent Pat Biotechnol, 2009, 3(2):141-153.
[2] Croxatto H B. Progestin implants. Steroids, 2000, 65(10-11):681-685.
[3] Hughes D T, Sperandio V. Inter-kingdom signalling:communication between bacteria and their hosts. Nat Rev Microbiol, 2008, 6(2):111-120.
[4] Bragin J, Saowakhon S, Manosroi A. A novel one-step biotransformation of cortexolone-21-acetate to hydrocortisone acetate using Cunninghamella blakesleeana ATCC 8688a. Enzyme Microb Technol, 2007, 41(3):322-325.
[5] Funder J W. Minireview:aldosterone and mineralocorticoid receptors:past, present, and future. Endocrinology, 2010, 151(11):5098-5102.
[6] García J L, Uhía I, Galán B. Catabolism and biotechnological applications of cholesterol degrading bacteria. Microb Biotechnol, 2012, 5(6):679-699.
[7] Bragin E Y, Shtratnikova V Y, Dovbnya D V, et al. Comparative analysis of genes encoding key steroid core oxidation enzymes in fast-growing Mycobacterium spp. strains. J Steroid Biochem Mol Biol, 2013, 138(10):41-53.
[8] Xie R, Shen Y, Qin N, et al. Genetic differences in ksdD influence on the ADD/AD ratio of Mycobacterium neoaurum. J Ind Microbiol Biotechnol, 2015, 42(4):507-513.
[9] Cruz A, Angelova B, Fernandes P, et al. Study of key operational parameters for the side-chain cleavage of sitosterol by free mycobacterial cells in bis-(2-ethylhexyl) phthalate. Biocatal Biotransform, 2004, 22(3):189-194.
[10] Fokina V V, Sukhodol'skaya G V, Gulevskaya S A, et al. The 1(2)-dehydrogenation of steroid substrates by Nocardioides simplex VKM Ac-2033D. Microbiology, 2003, 72(1):24-29.
[11] Fokina V V, Donova M V. 21-Acetoxy-pregna-4(5),9(11),16(17)-triene-21-ol-3,20-dione conversion by Nocardioides simplex VKM Ac-2033D. J Steroid Biochem Mol Biol, 2003, 87(4-5):319-325.
[12] Donova M V. Transformation of steroids by actinobacteria:a review. Applied Biochemistry and Microbiology, 2007, 43(1):5-18.
[13] Donova M V, Egorova O V. Microbial steroid transformations:Current state and prospects. Appl Microbiol Biotechnol, 2012, 94(6):1423-1447.
[14] Fernandes P, Cruz A, Angelova B, et al. Microbial conversion of steroid compounds:Recent developments. Enzyme Microb Technol, 2003, 32(6):688-705.
[15] Wang F Q, Yao K, Xu L Q, et al. Characterization and engineering of 3-ketosteroid-△1-dehydrogenase and 3-ketosteroid-9α-hydroxylase in Mycobacterium neoaurum ATCC 25795 to produce 9α-hydroxy-4-androstene-3, 17-dione through the catabolism of sterols. Metabolic Engineering, 2014, 24:181-191.
[16] Wang F Q, Wei W, Fan S Y, et al. Inactivation and augmentation of the primary 3-ketosteroid-Δ1-dehydrogenase in Mycobacterium neoaurum NwIB-01:biotransformation of soybean phytosterols to 4-androstene-3, 17-dione or 1, 4-androstadiene-3, 17-dione. Appl Environ Microbiol, 2010, 76(13):4578-4582.
[17] Zhang W Q, Shao M L, Rao Z M, et al. Bioconversion of 4-androstene-3,17-dione toandrost-1,4-diene-3,17-dione by recombinant Bacillus subtilis expressing ksdd gene encoding 3-ketosteroid-Δ1-dehydrogenasefrom Mycobacterium neoaurum JC-12. J Steroid Biochem MolBiol, 2013, 135(1):36-42.
[18] Li Y, Lu F, Sun T, et al. Expression of ksdD gene encoding 3-ketosteroid-Δ1-dehydrogenase from Arthrobacter simplex in Bacillus subtilis. Lett Appl Microbiol, 2007, 44(5):563-568.
[19] Malaviya A, Gomes J. Androstenedione production by biotransformation of phytosterols. Bioresource Technology, 2008, 99(15):6725-6737.
[20] Shao M, Zhang X, Rao Z, et al. Enhanced production of androst-1,4-diene-3,17-dione by Mycobacterium neoaurum JC-12 using three-stage fermentation strategy. PLoS One, 2015, 10(9):e0137658.
[21] Flett F, Mersinias V, Smith C P. High efficiency intergeneric conjugal transfer of plasmid DNA from Escherichia coli to methyl DNA-restricting streptomycetes. FEMS Microbiol Lett, 1997, 155(2):223-229.
[22] Van D G, Hessels G I, Van G R, et al. Targeted disruption of the kstD geneencoding 3-ketosteroid-Δ1-dehydrogenase isoenzyme of Rhodococcus erythropolis SQ1. Appl Environ Microbiol, 2000, 66(5):2029-2036. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
