Annals of Microbiology

, Volume 65, Issue 4, pp 1961–1971 | Cite as

Multiplicity of 3-ketosteroid Δ1-dehydrogenase enzymes in Gordonia neofelifaecis NRRL B-59395 with preferences for different steroids

  • Qingyan Zhang
  • Yao Ren
  • Junzhong He
  • Shijun Cheng
  • Jiadai Yuan
  • Fanglan Ge
  • Wei Li
  • Ying Zhang
  • Gangrong Xie
Original Article
First Online:
Received:
Accepted:

Abstract

The presence of redundant genes in a genome might enable bacteria to adapt to varying environmental conditions. The genus Gordonia comprises metabolically diverse bacteria that can degrade a wide range of persistent compounds. This paper reports the multiplicity of genes encoding 3-ketosteroid Δ1-dehydrogenase (KstD) in Gordonia neofelifaecis NRRL B-59395. KstD is one of the key enzymes in microbial steroid catabolism. Five kstD homologues (kstD1–kstD5) distributed over several phylogenetic groups were investigated in G. neofelifaecis NRRL B-59395. Escherichia coli cells expressing kstD homologues were used to transform different steroids, including cholesterol, cholest-4-en-3-one, 4-androstene-3,17-dione (AD), progesterone, and 16α,17α-epoxyprogesterone. All five enzymes displayed KstD activity toward 3-ketosteroids, but none were able to transform 3-hydroxysteroid substrates such as cholesterol. KstD2, KstD3, and KstD5 catalyzed the dehydrogenation reaction of 16α,17α-epoxyprogesterone, while KstD1, KstD3, and KstD4 transformed AD into androsta-1,4-diene-3,17-dione. Transcriptional analyses revealed that the expression of kstD1, kstD3, and kstD4 was upregulated during culture in cholesterol or AD when compared with expression during culture in pyruvic acid. kstD2 and kstD5 were uniquely induced by cholesterol. Thus, the presence of multiple kstD genes in the G. neofelifaecis NRRL B-59395 genome facilitates a dynamic and fine-tuned response to environmental changes.

Keywords

Gordonia neofelifaecis 3-ketosteroid Δ1-dehydrogenase Steroids Transformation 
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Notes

Acknowledgments

The research was funded by the National Natural Science Foundation of China (No. 31271332), the Sichuan provincial Science & Technology Department (2008JY0103-1, 2009JY0067), and the Sichuan provincial Department of Education (13ZA0145). We thank the Beijing Genomics Institute at Shenzhen for technical assistance with the RNA-Seq data analysis.

References

  1. Brzezinska M, Szulc I, Brzostek A, Klink M, Kielbik M, Sulowska Z, Pawelczyk J, Dziadek J (2013) The role of 3-ketosteroid 1(2)-dehydrogenase in the pathogenicity of Mycobacterium tuberculosis. BMC Microbiol 20:13–43Google Scholar
  2. Brzostek A, Sliwiński T, Rumijowska-Galewicz A, Korycka-Machała M, Dziadek J (2005) Identification and targeted disruption of the gene encoding the main 3-ketosteroid dehydrogenase in Mycobacterium smegmatis. Microbiology 151(Pt 7):2393–23402CrossRefPubMedGoogle Scholar
  3. Burns G, Brown T, Hatter K, Sokatch JR (1989) Sequence analysis of the lpdV gene for lipoamide dehydrogenase of branched-chain-oxoacid dehydrogenase of Pseudomonas putida. Eur J Biochem 179:61–69CrossRefPubMedGoogle Scholar
  4. Chiang YR, Ismail W, Gallien S, Heintz D, Van Dorsselaer A, Fuchs G (2008) Cholest-4-en-3-one-delta 1-dehydrogenase, a flavoprotein catalyzing the second step in anoxic cholesterol metabolism. Appl Environ Microbiol 74:107–113PubMedCentralCrossRefPubMedGoogle Scholar
  5. Choi KP, Molnár I, Yamashita M, Murooka Y (1995) Purification and characterization of the 3-ketosteroid-Δ1-dehydrogenase of Arthrobacter simplex produced in Streptomyces lividans. J Biochem 117(5):1043–1049PubMedGoogle Scholar
  6. Donova MV (2007) Transformation of steroids by actinobacteria: a review. Appl Biochem Microbiol 43:1–14CrossRefGoogle Scholar
  7. Fernández de las Heras L, van der Geize R, Drzyzga O, Perera J, María Navarro Llorens J (2012) Molecular characterization of three 3-ketosteroid-Δ(1)-dehydrogenase isoenzymes of Rhodococcus ruber strain Chol-4. J Steroid Biochem Mol Biol 132:271–281CrossRefPubMedGoogle Scholar
  8. Florin C, Köhler T, Grandguillot M, Plesiat P (1996) Comamonas testosteroni 3-ketosteroid-delta 4(5 alpha)-dehydrogenase: gene and protein characterization. J Bacteriol 178:3322–3330PubMedCentralPubMedGoogle Scholar
  9. García JL, Uhía I, Galán B (2012) Catabolism and biotechnological applications of cholesterol degrading bacteria. Microb Biotechnol 5:679–699PubMedCentralCrossRefPubMedGoogle Scholar
  10. Ge F, Li W, Chen G, Liu Y, Zhang G, Yong B, Wang Q, Wang N, Huang Z, Li W, Wang J, Wu C, Xie Q, Liu G (2011) Draft genome sequence of G. neofelifaecis NRRL B-59395, a cholest-4-en-3-one-degrading actinomycete. J Bacteriol 193:5045–5046PubMedCentralCrossRefPubMedGoogle Scholar
  11. Hiessl S, Schuldes J, Thürmer A, Halbsguth T, Bröker D, Angelov A, Liebl W, Daniel R, Steinbüchel A (2012) Involvement of two latex-clearing proteins during rubber degradation and insights into the subsequent degradation pathway revealed by the genome sequence of Gordonia polyisoprenivorans strain VH2. Appl Environ Microbiol 78:2874–2887PubMedCentralCrossRefPubMedGoogle Scholar
  12. Kieslich K (1985) Microbial side-chain degradation of sterols. J Basic Microbiol 25:461–474CrossRefPubMedGoogle Scholar
  13. Knol J, Bodewits K, Hessels GI, Dijkhuizen L, van der Geize R (2008) 3-Keto-5alpha-steroid Delta(1)-dehydrogenase from Rhodococcus erythropolis SQ1 and its ortholog in Mycobacterium tuberculosis H37Rv are highly specific enzymes that function in cholest-4-en-3-one catabolism. Biochem J 410:339–346CrossRefPubMedGoogle Scholar
  14. Linos A, Steinbüchel A, Spröer C, Kroppenstedt RM (1999) Gordonia polyisoprenivorans sp. nov., a rubber-degrading actinomycete isolated from an automobile tyre. Int J Syst Bacteriol 49:1785–1791CrossRefPubMedGoogle Scholar
  15. Liu Y, Chen YG, Ge F, Li W, Zeng L, Cao WG (2011a) Efficient biotransformation of cholest-4-en-3-one to androsta-1,4-diene-3, 17-dione by a newly isolated actinomycete Gordonia neofelifaecis. W J Microbiol Biotechnol 27:4759–4765Google Scholar
  16. Liu Y, Ge F, Chen G, Li W, Ma P, Zhang G, Zeng L (2011b) Gordonia neofelifaecis sp. nov., a cholest-4-en-3-one side-chain cleaving actinomycete isolated from the faeces of Neofelis nebulosa. Int J Syst Evol Mircobiol 61:165–169CrossRefGoogle Scholar
  17. Malaviya A, Gomes J (2008) Androstenedione production by biotransformation of phytosterols. Bioresour Technol 99(15):6725–6737CrossRefPubMedGoogle Scholar
  18. Owen RW, Mason AN, Bilton RF (1983) The degradation of cholesterol by Pseudomonas sp. NCIB 10590 under aerobic conditions. J Lipid Res 24:1500–1511PubMedGoogle Scholar
  19. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 1228–1231Google Scholar
  20. Uhía I, Galán B, Kendall SL, Stoker NG, García JL (2012) cholest-4-en-3-one metabolism in Mycobacterium smegmatis. Environ Microbiol Rep 4:168–182CrossRefPubMedGoogle Scholar
  21. Wei W, Wang FQ, Fan SY, Wei DZ (2010) Inactivation and augmentation of the primary 3-ketosteroid-{delta}1- 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 76:4578–4582PubMedCentralCrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg and the University of Milan 2015

Authors and Affiliations

  • Qingyan Zhang
    • 1
  • Yao Ren
    • 1
  • Junzhong He
    • 1
  • Shijun Cheng
    • 1
  • Jiadai Yuan
    • 1
  • Fanglan Ge
    • 1
  • Wei Li
    • 1
  • Ying Zhang
    • 1
  • Gangrong Xie
    • 1
  1. 1.College of Life SciencesSichuan Normal UniversityChengduPeople’s Republic of China

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