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2,3-Butanediol production by the non-pathogenic bacterium Paenibacillus brasilensis

  • Biotechnological products and process engineering
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Abstract

2,3-Butanediol (2,3-BDO) is of considerable importance in the chemical, plastic, pharmaceutical, cosmetic, and food industries. The main bacterial species producing this compound are considered pathogenic, hindering large-scale productivity. The species Paenibacillus brasilensis is generally recognized as safe (GRAS) and is phylogenetically similar to P. polymyxa, a species widely used for 2,3-BDO production. Here, we demonstrate, for the first time, that P. brasilensis strains produce 2,3-BDO. Total 2,3-BDO concentrations for 15 P. brasilensis strains varied from 5.5 to 7.6 g/l after 8 h incubation at 32 °C in modified YEPD medium containing 20 g/l glucose. Strain PB24 produced 8.2 g/l of 2,3-BDO within a 12-h growth period, representing a yield of 0.43 g/g and a productivity of 0.68 g/l/h. An increase in 2,3-BDO production by strain PB24 was observed using higher concentrations of glucose, reaching 27 g/l of total 2,3-BDO in YEPD containing about 80 g/l glucose within a 72-h growth period. We sequenced the genome of P. brasilensis PB24 and uncovered at least six genes related to the 2,3-BDO pathway at four distinct loci. We also compared gene sequences related to the 2,3-BDO pathway in P. brasilensis PB24 with those of other spore-forming bacteria, and found strong similarity to P. polymyxa, P. terrae, and P. peoriae 2,3-BDO-related genes. Regulatory regions upstream of these genes indicated that they are probably co-regulated. Finally, we propose a production pathway from glucose to 2,3-BDO in P. brasilensis PB24. Although the gene encoding S-2,3-butanediol dehydrogenase (butA) was found in the genome of P. brasilensis PB24, only R,R-2,3- and meso-2,3-butanediol were detected by gas chromatography under the growth conditions tested here. Our findings can serve as a basis for further improvements to the metabolic capabilities of this little-studied Paenibacillus species in relation to production of the high-value chemical 2,3-butanediol.

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References

  • Adlakha N, Pfau T, Ebenhöh O, Yazdani SS (2015) Insight into metabolic pathways of the potential biofuel producer, Paenibacillus polymyxa ICGEB2008. Biotechnol Biofuels 8:159

    Article  PubMed  PubMed Central  Google Scholar 

  • Adlakha N, Yazdani SS (2015) Efficient production of (R,R)-2,3-butanediol from cellulosic hydrolysate using Paenibacillus polymyxa ICGEB2008. J Ind Microbiol Biotechnol 42:21–28

    Article  CAS  PubMed  Google Scholar 

  • Aziz RK, Bartels D, Best AA, DeJongh M, Disz T, Edwards RA, Formsma K, Gerdes S, Glass EM, Kubal M, Meyer F, Olsen GJ, Olson R, Osterman AL, Overbeek RA, McNeil LK, Paarmann D, Paczian T, Parrello B, Pusch GD, Reich C, Stevens R, Vassieva O, Vonstein V, Wilke A, Zagnitko O (2008) The RAST server: rapid annotations using subsystems technology. BMC Genomics 9:75

    Article  PubMed  PubMed Central  Google Scholar 

  • Bailey TL, Gribskov M (1998) Combining evidence using p-values: application to sequence homology searches. Bioinformatics 14:48–54

    Article  CAS  PubMed  Google Scholar 

  • Celinska E, Grajek W (2009) Biotechnological production of 2,3-butanediol–current state and prospects. Biotechnol Adv 27:715–725

    Article  CAS  PubMed  Google Scholar 

  • Chin C-S, Alexander DH, Marks P, Klammer AA, Drake J, Heiner C, Clum A, Copeland A, Huddleston J, Eichler EE, Turner SW, Korlach J (2013) Nonhybrid, finished microbial genome assemblies from long-read SMRT sequencing data. Nat Methods 10:563–569

    Article  CAS  PubMed  Google Scholar 

  • Cho S, Kim T, Woo HM, Lee J, Kim Y, Um Y (2015) Enhanced 2, 3-butanediol production by optimizing fermentation conditions and engineering Klebsiella oxytoca M1 through overexpression of acetoin reductase. PLoS One 10:e0138109

    Article  PubMed  PubMed Central  Google Scholar 

  • Fortes TO, Alviano DS, Tupinambá G, Padrón TS, Antoniolli AR, Alviano CS, Seldin L (2008) Production of an antimicrobial substance against Cryptococcus neoformans by Paenibacillus brasilensis Sa3 isolated from the rhizosphere of Kalanchoe brasiliensis. Microbiol Res 163:200–207

    Article  PubMed  Google Scholar 

  • Garg SK, Jain A (1995) Fermentative production of 2,3-butanediol—a review. Bioresour Technol 51:103–109

    Article  CAS  Google Scholar 

  • González E, Fernández MR, Larroy C, Solà L, Pericàs MA, Parés X, Biosca JA (2000) Characterization of a (2R,3R)-2,3-butanediol dehydrogenase as the Saccharomyces cerevisiae YAL060W gene product. Disruption and induction of the gene J Biol Chem 275:35876–33585

  • Hammer Ø, Harper DAT, Ryan PD (2001) Paleontological statistics software package for education and data analysis. Palaeontol Electron 4:9–18

    Google Scholar 

  • Ji XJ, Huang H, Du J, Zhu JG, Ren LJ, Hu N, Li S (2009) Enhanced 2,3-butanediol production by Klebsiella oxytoca using a two-stage agitation speed control strategy. Bioresour Technol 100:3410–3414

    Article  CAS  PubMed  Google Scholar 

  • Ji XJ, Huang H, Ouyang PK (2011) Microbial 2,3-butanediol production: a state-of-the-art review. Biotechnol Adv 29:351–364

    Article  CAS  PubMed  Google Scholar 

  • Jiang L-q, Fang Z, Zaho Z-l, He F, Li H-b (2015) 2,3-Butanediol and acetoin production from enzymatic hydrolysate of ionic liquid-pretreated cellulose by Paenibacillus polymyxa. Bioresources 10:1318–1329

    Google Scholar 

  • Kallbach M, Horn S, Kuenz A, Prüße U (2017) Screening of novel bacteria for the 2,3-butanediol production. Appl Microbiol Biotechnol 101:1025–1033

    Article  CAS  PubMed  Google Scholar 

  • Köpke M, Mihalcea C, Liew F, Tizard JH, Ali MS, Conolly JJ, Al-Sinawi B, Simpson SD (2011) 2,3-butanediol production by acetogenic bacteria, an alternative route to chemical synthesis, using industrial waste gas. Appl Environ Microbiol 77:5467–5475

    Article  PubMed  PubMed Central  Google Scholar 

  • Kumar S, Stecher G, Tamura K (2016) MEGA7: molecular evolutionary genetics analysis version 7.0 for bigger datasets. Mol Biol Evol 33:1870–1874

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948

    Article  CAS  PubMed  Google Scholar 

  • Meyer F, Overbeek R, Rodriguez A (2009) FIGfams: yet another set of protein families. Nucleic Acids Res 37:6643–6654

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Moriya Y, Itoh M, Okuda S, Yoshizawa A, Kanehisa M (2007) KAAS: an automatic genome annotation and pathway reconstruction server. Nucleic Acids Res 35:182–185

    Article  Google Scholar 

  • Okonkwo CC, Ujor VC, Mishra PK, Ezeji TC (2017) Process development for enhanced 2,3-butanediol production by Paenibacillus polymyxa DSM 365. Fermentation 3:18

    Article  Google Scholar 

  • Overbeek R, Olson R, Pusch GD, Olsen GJ, Davis JJ, Disz T, Edwards RA, Gerdes S, Parrello B, Shukla M, Vonstein V, Wattam AR, Xia F, Stevens R (2014) The SEED and the rapid annotation of microbial genomes using subsystems technology (RAST). Nucleic Acids Res 42:D206–D214

    Article  CAS  PubMed  Google Scholar 

  • Overmars L, Kerkhoven R, Siezen RJ, Francke C (2013) MGcV: the microbial genomic context viewer for comparative genome analysis. BMC Genomics 14:209

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Petrov K, Petrova P (2009) High production of 2,3-butanediol from glycerol by Klebsiella pneumoniae G31. Appl Microbiol Biotechnol 84:659–665

    Article  CAS  PubMed  Google Scholar 

  • Qiu Y, Zhang J, Li L, Wen Z, Nomura CT, Wu S, Chen S (2016) Engineering Bacillus licheniformis for the production of meso-2,3-butanediol. Biotechnol Biofuels 9:117

    Article  PubMed  PubMed Central  Google Scholar 

  • Sabra W, Groeger C, Zeng AP (2015) Microbial cell factories for diol production. Adv Biochem Eng Biotechnol 155:165–197

    Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Seldin L, Dubnau D (1985) Deoxyribonucleic acid homology among Bacillus polymyxa, Bacillus macerans, Bacillus azotofixans and other nitrogen-fixing Bacillus strains. Int J Syst Bacteriol 35:151–154

    Article  CAS  Google Scholar 

  • Seldin L, Penido EGC (1986) Identification of Bacillus azotofixans using API tests. Antonie Van Leeuwenhoek 52:403–409

    Article  CAS  PubMed  Google Scholar 

  • Seldin L, Rosado AS, Cruz DW, Nobrega A, van Elsas JD, Paiva E (1998) Comparison of Paenibacillus azotofixans strains isolated from rhizoplane, rhizosphere and non-rhizosphere soil from maize planted in two different Brazilian soils. Appl Environ Microbiol 64:3860–3868

    CAS  PubMed  PubMed Central  Google Scholar 

  • Song CW, Rathnasingh C, Park JM, Lee J, Hyohak Song H (2018) Isolation and evaluation of Bacillus strains for industrial production of 2,3-butanediol. J Microbiol Biotechnol 28:409–417

    Article  PubMed  Google Scholar 

  • Syu MJ (2001) Biological production of 2,3-butanediol. Appl Microbiol Biotechnol 55:10–18

    Article  CAS  PubMed  Google Scholar 

  • US FDA/CFSAN (2015) Inventory of GRAS notices: summary of all GRAS notices. https://www.fda.gov/food/ingredientspackaginglabeling/gras/; accessed in March 2015

  • von der Weid I, Artursson V, Seldin L, Jansson JK (2005) Antifungal and root surface colonization properties of GFP-tagged Paenibacillus brasilensis PB177. World J Microbiol Biotechnol 21:1591–1597

    Article  Google Scholar 

  • von der Weid I, Duarte GF, van Elsas JD, Seldin L (2002) Paenibacillus brasilensis sp. nov., a novel nitrogen-fixing species isolated from the maize rhizosphere in Brazil. Int J Syst Evol Microbiol 52:2147–2153

    PubMed  Google Scholar 

  • Yang T, Rao Z, Zhang X, Xu M, Xu Z, Yang ST (2017) Metabolic engineering strategies for acetoin and 2, 3-butanediol production: advances and prospects. Crit Rev Biotechnol 37:990–1005

    Article  PubMed  Google Scholar 

  • Yu B, Sun J, Bommareddy RR, Song L, Zeng A-P (2011) Novel (2R,3R)-2,3-butanediol dehydrogenase from potential industrial strain Paenibacillus polymyxa ATCC 12321. Appl Environ Microbiol 77:4230–4233

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhang L, Guo Z, Chen J, Xu Q, Lin H, Hu K, Guan X, Shen Y (2016) Mechanism of 2,3-butanediol stereoisomers formation in a newly isolated Serratia sp. T241. Sci Rep 6:19257

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

Thanks are due to Fábio Diniz for his technical support in the HPLC analyses. We are grateful to Prof. Apostolis Koutinas and Dr. Chrysanthi Pateraki from the Agricultural University of Athens, Attiki, Greece, for their helpful comments and guidance.

Funding

This study was supported by grants from the National Research Council of Brazil (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) and Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ). M.E.V. and D.M.G.F. were provided a grant by Petrobras (grant number: 2012/00320-2).

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Author notes

  1. Beatriz do Carmo Dias and Maria Eduarda do Nascimento Vitorino Lima contributed equally to this work.

Authors and Affiliations

  1. Laboratório de Genética Microbiana, Departamento de Microbiologia Geral, Instituto de Microbiologia Paulo de Góes, Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde, Bloco I, Ilha do Fundão, Rio de Janeiro, RJ, CEP 21941.902, Brazil

    Beatriz do Carmo Dias, Renata Estebanez Vollú & Lucy Seldin

  2. Instituto de Química, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, 21941-909, Brazil

    Maria Eduarda do Nascimento Vitorino Lima & Denise Maria Guimarães Freire

  3. Instituto Oswaldo Cruz, Fiocruz, Rio de Janeiro, RJ, 21040-900, Brazil

    Fabio Faria da Mota

  4. Instituto de Pesquisas de Produtos Naturais, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, 21941-902, Brazil

    Antonio Jorge Ribeiro da Silva

  5. PETROBRAS R&D Center/Biotechnology, Rio de Janeiro, RJ, 21941-915, Brazil

    Aline Machado de Castro

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  1. Beatriz do Carmo Dias
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  2. Maria Eduarda do Nascimento Vitorino Lima
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Correspondence to Lucy Seldin.

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Dias, B.d., Lima, M.E.d., Vollú, R.E. et al. 2,3-Butanediol production by the non-pathogenic bacterium Paenibacillus brasilensis. Appl Microbiol Biotechnol 102, 8773–8782 (2018). https://doi.org/10.1007/s00253-018-9312-y

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