Advertisement

Applied Microbiology and Biotechnology

, Volume 103, Issue 1, pp 339–348 | Cite as

Selective and faster nicotine biodegradation by genetically modified Pseudomonas sp. JY-Q in the presence of glucose

  • Hui Zhang
  • Rui Zhao
  • Chaochao Huang
  • Jun Li
  • Yunhai Shao
  • Jian Xu
  • Ming Shu
  • Weihong Zhong
Applied microbial and cell physiology

Abstract

Pseudomonas sp. JY-Q is a nicotine-degrading strain isolated from tobacco waste extract (TWE). In TWE, the nicotine is a toxic chemical and requires removal. However, it was found that glucose in TWE inhibited the degradation of nicotine. Bioinformatics analysis of JY-Q complete genome found five genes encoding the first-step enzymes of glucose metabolism, one glucokinase (gck, AA098_22370) and four glucose dehydrogenases (gdh, AA098_12490, 22860, 11910, and 05800). Homogonous recombinant strategy was utilized to delete all the five genes from JY-Q genome one by one. The resultant quinary mutant strain JY-Q/5∆ exhibited no growth on glucose as the sole carbon source and selective degradation of nicotine in medium coexisting with glucose. The result of single complementation in the quinary mutant showed that only gck and gdh-05800 genes exhibited significant effect on the initial steps of glucose metabolism. Although the growth of JY-Q/5∆ seemed worse in basic inorganic medium (BSM) with coexisting glucose and nicotine, the nicotine degradation rate per cell weight of JY-Q/5∆ reached 12.68 mg/mg/h, about four times higher than that of the wild-type strain. The resting cells of JY-Q/5∆ also showed better ability of nicotine degradation than the wild type in BSM coexisting with glucose. In 5% diluted TWE containing 0.8 g/L nicotine, the resting cells of JY-Q/5∆ degraded all nicotine within 24 h, 20% faster than the wild-type strain. JY-Q/5∆ is potential to selectively degrade nicotine in glucose-nicotine coexisting environment.

Keywords

Nicotine Glucose inhibition Selectively biodegradation Gene knock out Tobacoo waste extract Pseudomonas sp. JY-Q 

Notes

Acknowledgements

This study was supported by the National Natural Science Foundation of China (31670115). Professor Ningyi Zhou of Shanghai Jiaotong University was also appreciated for his helpful suggestions.

Compliance with ethical standards

The authors declare that they comply with ethical standards.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Supplementary material

253_2018_9445_MOESM1_ESM.pdf (228 kb)
ESM 1 (PDF 227 kb)

References

  1. Baitsch D, Sandu C, Brandsch R, Igloi GL (2001) Gene cluster on pAO1 of Arthrobacter nicotinovorans involved in degradation of the plant alkaloid nicotine: cloning purification and characterization of 2,6-dihydroxypyridine 3-hydroxylase. J Bacteriol 183:5262–5267CrossRefGoogle Scholar
  2. Boopathy R (2000) Factors limiting bioremediation technologies. Bioresour Technol 74:63–67CrossRefGoogle Scholar
  3. Brandsch R (2006) Microbiology and biochemistry of nicotine degradation. Appl Microbiol Biotechnol 69:493–498CrossRefGoogle Scholar
  4. Brückner R, Titgemeyer F (2002) Carbon catabolite repression in bacteria: choice of the carbon source and autoregulatory limitation of sugar utilization. FEMS Microbiol Lett 209:141–148CrossRefGoogle Scholar
  5. Cadoret F, Soscia C, Voulhoux R (2014) Gene transfer: transformation/electroporation. Methods Mol Biol 1149:11–15CrossRefGoogle Scholar
  6. Cai YQ, Qi XN, Qi Q, Lin YP, Wang ZX, Wang QH (2018) Effect of MIG1 and SNF1 deletion on simultaneous utilization of glucose and xylose by Saccharomyces cerevisiae. Chin J Biotechnol 34:54–67Google Scholar
  7. del Castillo T, Ramos JL (2007) Simultaneous catabolite repression between glucose and toluene metabolism in Pseudomonas putida is channeled through different signaling pathways. J Bacteriol 189:6602–6610CrossRefGoogle Scholar
  8. del Castillo T, Ramos JL, Rodriguez-Herva JJ, Fuhrer T, Sauer U, Duque E (2007) Convergent peripheral pathways catalyze initial glucose catabolism in Pseudomonas putida: genomic and flux analysis. J Bacteriol 189:5142–5152CrossRefGoogle Scholar
  9. Galinier A (2018) Carbon catabolite repression or how bacteria choose their favorite sugars. Med Sci 34:531–539Google Scholar
  10. Huang HY, Yu WJ, Wang RS, Li HL, Xie HJ, Wang SN (2017) Genomic and transcriptomic analyses of Agrobacterium tumefaciens S33 reveal the molecular mechanism of a novel hybrid nicotine-degrading pathway. Sci Rep 7:4813CrossRefGoogle Scholar
  11. Ishige T, Honda K, Shimizu S (2005) Whole organism biocatalysis. Curr Opin Chem Biol 9:174–180CrossRefGoogle Scholar
  12. Kovach ME, Elzer PH, Hill DS, Robertson GT, Farris MA, Roop RM, Peterson KM (1995) Four new derivatives of the broad-host-range cloning vector pBBR1MCS, carrying different antibiotic-resistance cassettes. Gene 166:175–176CrossRefGoogle Scholar
  13. Li J, Qian SL, Xiong L, Zhu CY, Shu M, Wang J, Jiao Y, He HL, Zhang FM, Linhardt RJ, Zhong WH (2017) Comparative genomics reveals specific genetic architectures in nicotine metabolism of pseudomonas sp. JY-Q. Front Microbiol 8:2085CrossRefGoogle Scholar
  14. Liu HG, He HL, Cheng CH, Liu JL, Shu M, JiaoY TF, Zhong WH (2015a) Diversity analysis of the bacterial community in tobacco waste extract during reconstituted tobacco process. Appl Microbiol Biotechnol 99:469–476CrossRefGoogle Scholar
  15. Liu JL, Ma GH, Chen T, Hou Y, Yang SH, Zhang KQ, Yang JK (2015b) Nicotine-degrading microorganisms and their potential applications. Appl Microbiol Biotechnol 99:3775–3785CrossRefGoogle Scholar
  16. Lynch SA, Gill RT (2012) Synthetic biology: new strategies for directing design. Metab Eng 14:205–211CrossRefGoogle Scholar
  17. Ma Y, Wei Y, Qiu JG, Wen RT, Hong J, Liu WP (2014) Isolation, transposon mutagenesis, and characterization of the novel nicotine-degrading strain Shinella sp. HZN7. Appl Microbiol Biotechnol 98:2625–2636CrossRefGoogle Scholar
  18. Mustafa G, Ishikawa Y, Kobayashi K, Migita CT, Elias MD, Nakamura S, Tagawa S, Yamada M (2008) Amino acid residues interacting with both the bound quinone and coenzyme, pyrroloquinoline quinone, in Escherichia coli membrane-bound glucose dehydrogenase. J Biol Chem 283:22215–22221CrossRefGoogle Scholar
  19. Newhouse P, Singh A, Potter A (2004) Nicotine and nicotinic receptor involvement in neuropsychiatric disorders. Curr Top Med Chem 4:267–282CrossRefGoogle Scholar
  20. Novotny TE, Zhao F (1999) Consumption and production waste: another externality of tobacco use. Tob Control 8:75–80CrossRefGoogle Scholar
  21. Piotrowska-Cyplik A, Olejnik A, Cyplik P, Dach J, Czarnecki Z (2009) The kinetics of nicotine degradation, enzyme activities and genotoxic potential in the characterization of tobacco waste composting. Bioresour Technol 100:5037–5044CrossRefGoogle Scholar
  22. Qiu JG, Yang YJ, Zhang JJ, Wang HX, Ma Y, He J, Lu ZM (2016) The complete genome sequence of the nicotine-degrading bacterium Shinella sp. HZN7. Front Microbiol 7:1348CrossRefGoogle Scholar
  23. Quandt J, Hynes MF (1993) Versatile suicide vectors which allow direct selection for gene replacement in gram-negative bacteria. Gene 127:15–21CrossRefGoogle Scholar
  24. Ramakrishnan GG, Nehru G, Suppuram P, Balasubramaniyam S, Gulab BR, Subramanian R (2015) Bio-transformation of glycerol to 3-hydroxypropionic acid using resting cells of Lactobacillus reuteri. Curr Microbiol 71:517–523CrossRefGoogle Scholar
  25. Roca C, Haack MB, Olsson L (2004) Engineering of carbon catabolite repressionin recombinant xylose fermenting Saccharomyces cerevisiae. Appl Microbiol Biotechnol 63:578–583CrossRefGoogle Scholar
  26. Roduit JP, Wellig A, Kiener A (1997) Renewable functionalized pyridines derived from microbial metabolites of the alkaloid (S)-nicotine. Heterocycles 45:1687–1702CrossRefGoogle Scholar
  27. Schafer A, Tauch A, Jager W, Kalinowski J, Thierbach G, Puhler A (1994) Small mobilizable multi-purpose cloning vectors derived from the Escherichia coli plasmids pK18 and pK19: selection of defined deletions in the chromosome of Corynebacterium glutamicum. Gene 145:69–73CrossRefGoogle Scholar
  28. Wang JH, He HZ, Wang MZ, Wang S, Zhang J, Wei W, Xu HX, Lv ZM, Shen DS (2013) Bioaugmentation of activated sludge with Acinetobacter sp. TW enhances nicotine degradation in a synthetic tobacco wastewater treatment system. Bioresour Technol 142:445–453CrossRefGoogle Scholar
  29. Wang LJ, Tang HZ, Yu H, Yao YX, Xu P (2014) An unusual repressor controls the expression of a crucial nicotine-degrading gene cluster in Pseudomonas putida S16. Mol Microbiol 91:1252–1269CrossRefGoogle Scholar
  30. Wang WW, Xu P, Tang HZ (2015) Sustainable production of valuable compound 3-succinoyl-pyridine by genetically engineering Pseudomonas putida using the tobacco waste. Sci Rep 5:16411CrossRefGoogle Scholar
  31. Yin Y, Huang WY, Chen DJ (2007) Preparation of validoxylamine A by biotransformation of validamycin A using resting cells of a recombinant Escherichia coli. Biotechnol Lett 29:285–290CrossRefGoogle Scholar
  32. Yu H, Tang HZ, Wang LJ, Yao YX, Wu G, Xu P (2011) Complete genome sequence of the nicotine- degrading Pseudomonas putida strain S16. J Bacteriol 193:5541–5542CrossRefGoogle Scholar
  33. Yu H, Tang HZ, Zhu XY, Li YY, Xu P (2015) Molecular mechanism of nicotine degradation by a newly isolated strain, Ochrobactrum sp. strain SJY1. Appl Environ Microbiol 81:272–281CrossRefGoogle Scholar
  34. Yu WJ, Li HL, Xie KB, Huang HY, Xie HJ, Wang SN (2016) Genome sequence of the nicotine-degrading Agrobacterium tumefaciens S33. J Biotechnol 228:1–2CrossRefGoogle Scholar
  35. Yu WJ, Wang RS, Li HL, Liang JY, Wang YY, Huang HY, Xie HJ, Wang SN (2017) Green route to synthesis of valuable chemical 6-hydroxynicotine from nicotine in tobacco wastes using genetically engineered Agrobacterium tumefaciens S33. Biotechnol Biofuels 10:288CrossRefGoogle Scholar
  36. Zhong WH, Zhu CJ, Shu M, Sun KD, Zhao L, Wang C, Ye ZJ, Chen JM (2010) Degradation of nicotine in tobacco waste extract by newly isolated Pseudomonas sp. ZUTSKD. Bioresour Technol 101:6935–6941CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.College of Biotechnology and BioengineeringZhejiang University of TechnologyHangzhouPeople’s Republic of China
  2. 2.Technology Center, China Tobacco Zhejiang Industrial Co. Ltd.HangzhouPeople’s Republic of China

Personalised recommendations