Co-production of 1,3-propanediol and phage phiKpS2 from the glycerol fermentation by Klebsiella pneumoniae

Abstract

As an alternative to antibiotics in response to antimicrobial-resistant infections, bacteriophages (phages) are garnering renewed interest in recent years. However, the massive preparation of phage is restricted using traditional pathogens as host cells, which incurs additional costs and contamination. In this study, an opportunistic pathogen, Klebsiella pneumoniae used to convert glycerol to 1,3-propanediol (1,3-PDO), was reused to prepare phage after fermentation. The phage infection showed that the fed-batch fermentation broth containing 71.6 g/L 1,3-PDO can be directly used for preparation of phage with a titer of 1 × 10⁸ pfu/mL. Then, the two-step salting-out extraction was adopted to remove most impurities, e.g. acetic acid (93.5%), ethanol (91.5%) and cells (99.4%) at the first step, and obtain 1,3-PDO (56.6%) in the top phase as well as phage (97.4%) in the middle phase at the second step. This integrated process provides a cheap and environment-friendly manner for coproduction of 1,3-PDO and phage.

Keywords

Klebsiella pneumonia / 1,3-Propanediol / Bacteriophage / Co-production / Salting-out extraction

References

[1] Abedon ST. Lysis from without. Bacteriophage, 2011, 1(1): 46-49.

[2] Ames TT (2002) Process for the isolation of 1, 3-propanediol from fermentation broth. US Patent 6361983

[3] Asenjo JA, Andrews BA. Aqueous two-phase systems for protein separation: a perspective. J Chromatogr A, 2011, 1218(49): 8826-8835.

[4] Cabral J (2007) Cell partitioning in aqueous two-phase polymer systems. Cell Sep 151–171. https://doi.org/10.1007/10_2006_045

[5] Chan BK, Sistrom M, Wertz JE, Kortright KE, Narayan D, Turner PE. Phage selection restores antibiotic sensitivity in MDR Pseudomonas aeruginosa. Sci Rep, 2016, 6(1): 26717.

[6] Chen D, Dong YS, Bao YM, Xiu ZL (2023) Salting-out extraction of recombinant κ‐carrageenase and phage T7 released from Escherichia coli cells. Eng Life Sci 2200125. https://doi.org/10.1002/elsc.202200125

[7] Chilakamarry CR, Mimi Sakinah A, Zularism A, Khilji IA, Kumarasamy S (2022) Glycerol waste to bio-ethanol: Optimization of fermentation parameters by the Taguchi method. Journal of Chemistry 2022. https://doi.org/10.1155/2022/4892992

[8] Clavijo V, Torres-Acosta MA, Vives-Flórez MJ, Rito-Palomares M. Aqueous two-phase systems for the recovery and purification of phage therapy products: recovery of Salmonella bacteriophage ϕSan23 as a case study. Sep Purif Technol, 2019, 211: 322-329.

[9] Dong DX, Sutaria S, Hwangbo JY, Chen P. A simple and rapid method to isolate purer M13 phage by isoelectric precipitation. Appl Microbiol Biotechnol, 2013, 97(18): 8023-8029.

[10] Drab M. Phage aggregation–dispersion by ions: striving beyond Antibacterial Therapy. Trends Biotechnol, 2018, 36(9): 875-881.

[11] Duan S, Dong Y, Xu Y, Yin J, Geng L, Xiu Z. One-step salting-out extraction of bacteriophage from its infection broth of Acinetobacter baumannii. J Chromatogr A, 2022, 1679: 463407.

[12] Duan S, Ren L, Dong Y, Yang R, Xu Y, Yin J, Geng L, Xiu Z. Interfacial behavior and extraction kinetics of phages in a salting-out extraction system of ammonium citrate and ethyl acetate. J Ind Eng Chem, 2023

[13] Effah CY, Sun T, Liu S, Wu Y. Klebsiella pneumoniae: an increasing threat to public health. Ann Clin Microbiol Antimicrob, 2020, 19(1): 1-9.

[14] Gill JJ, Hyman P. Phage choice, isolation, and preparation for phage therapy. Curr Pharm Biotechnol, 2010, 11(1): 2-14.

[15] González-Mora A, Ruiz-Ruiz F, Benavides J, Willson RC, Rito-Palomares M. Recovery and primary purification of bacteriophage M13 using aqueous two-phase systems. J Chem Technol Biotechnol, 2017, 92(11): 2808-2816.

[16] Hassan KT, Ibraheem IJ, Hassan OM, Obaid A, Ali HH, Salih TA, Kadhim MS. Facile green synthesis of Ag/AgCl nanoparticles derived from Chara algae extract and evaluating their antibacterial activity and synergistic effect with antibiotics. J Environ Chem Eng, 2021, 9(4): 105359.

[17] Jacinto MJ, Patinha DJ, Marrucho IM, Gonçalves J, Willson RC, Azevedo AM, Aires-Barros MR. M13 bacteriophage purification using poly (ionic liquids) as alternative separation matrices. J Chromatogr A, 2018, 1532: 246-250.

[18] Jacinto M, Wagner A, Sá IM, Patinha DJ, Marrucho IM, Gonçalves J, Willson RC, Azevedo A, Aires-Barros M. Screening polymeric ionic liquids for chromatography-based purification of bacteriophage M13. Sep Purif Technol, 2021, 257: 117906.

[19] Jończyk-Matysiak E, Łodej N, Kula D, Owczarek B, Orwat F, Międzybrodzki R, Neuberg J, Bagińska N, Weber-Dąbrowska B, Górski A. Factors determining phage stability/activity: challenges in practical phage application. Expert Rev Anti Infect Ther, 2019, 17(8): 583-606.

[20] Lama S, Seol E, Park S. Development of Klebsiella pneumoniae J2B as microbial cell factory for the production of 1, 3-propanediol from glucose. Metab Eng, 2020, 62: 116-125.

[21] Lau GL, Sieo CC, Tan WS, Ho YW. Characteristics of a phage effective for colibacillosis control in poultry. J Sci Food Agric, 2012, 92(13): 2657-2663.

[22] Li Z, Jiang B, Zhang D, Xiu Z. Aqueous two-phase extraction of 1, 3-propanediol from glycerol-based fermentation broths. Sep Purif Technol, 2009, 66(3): 472-478.

[23] Li ZG, Jiang B, Zhang DJ, Xiu ZL. Aqueous two-phase extraction of 1,3-propanediol from glycerol-based fermentation broths. Sep Purif Technol, 2009, 66(3): 472-478.

[24] Li ZG, Teng H, Xiu ZL. Extraction of 1,3-propanediol from glycerol-based fermentation broths with methanol/phosphate aqueous two-phase system. Process Biochem, 2011, 46(2): 586-591.

[25] Li Z, Yan L, Zhou J, Wang X, Sun Y, Xiu Z-L. Two-step salting-out extraction of 1, 3-propanediol, butyric acid and acetic acid from fermentation broths. Sep Purif Technol, 2019, 209: 246-253.

[26] Li Z, Yan L, Zhou JJ, Wang XL, Sun YQ, Xiu ZL. Two-step salting-out extraction of 1,3-propanediol, butyric acid and acetic acid from fermentation broths. Sep Purif Technol, 2019, 209: 246-253.

[27] Liu HJ, Zhang DJ, Xu YH, Mu Y, Sun YQ, Xiu ZL. Microbial production of 1,3-propanediol from glycerol by Klebsiella pneumoniae under micro-aerobic conditions up to a pilot scale. Biotechnol Lett, 2007, 29(8): 1281-1285.

[28] Liu A, Liu Y, Peng L, Cai X, Shen L, Duan M, Ning Y, Liu S, Li C, Liu Y. Characterization of the narrow-spectrum bacteriophage LSE7621 towards Salmonella Enteritidis and its biocontrol potential on lettuce and tofu. LWT–Food Sci Technol, 2020, 118: 108791.

[29] Negrete A, Ling TC, Lyddiatt A. Aqueous two-phase recovery of bio-nanoparticles: a miniaturization study for the recovery of bacteriophage T4. J Chromatogr B, 2007, 854(1–2): 13-19.

[30] Oppermann S, Stein F, Kragl U. Ionic liquids for two-phase systems and their application for purification, extraction and biocatalysis. Appl Microbiol Biotechnol, 2011, 89(3): 493-499.

[31] Rabinovitch A, Aviram I, Zaritsky A. Bacterial debris—an ecological mechanism for coexistence of bacteria and their viruses. J Theor Biol, 2003, 224(3): 377-383.

[32] Raja S, Murty VR, Thivaharan V, Rajasekar V, Ramesh V. Aqueous two phase systems for the recovery of biomolecules–a review. Sci Technol, 2011, 1(1): 7-16.

[33] Salim A, Madhavan A, Babu P, Porayath C, Kesavan M, Hely S, Kumar VA, Nair BG, Pal S. Bacteriophage-based control of biogenic hydrogen sulphide produced by multidrug resistant Salmonella enterica in synthetic sewage. J Environ Chem Eng, 2021, 9(4): 105797.

[34] Serna-Galvis EA, Salazar-Ospina L, Jiménez JN, Pino NJ, Torres-Palma RA. Elimination of carbapenem resistant Klebsiella pneumoniae in water by UV-C, UV-C/persulfate and UV-C/H₂O₂. Evaluation of response to antibiotic, residual effect of the processes and removal of resistance gene. J Environ Chem Eng, 2020, 8(1): 102196.

[35] Shen GH, Wang JL, Wen FS, Chang KM, Kuo CF, Lin CH, Luo HR, Hung CH. Isolation and characterization of φkm18p, a novel lytic phage with therapeutic potential against extensively drug resistant Acinetobacter baumannii. PLoS ONE, 2012, 7(10): e46537.

[36] Shen JT, Zhou JJ, Fu HX, Mu Y, Sun YQ, Xu YP, Xiu ZL. A Klebsiella pneumoniae bacteriophage and its effect on 1,3-propanediol fermentation. Process Biochem, 2016, 51(10): 1323-1330.

[37] Song Z, Sun Y, Wei B, Xiu Z. Two-step salting‐out extraction of 1, 3‐propanediol and lactic acid from the fermentation broth of Klebsiella pneumoniae on biodiesel‐derived crude glycerol. Eng Life Sci, 2013, 13(5): 487-495.

[38] Szermer-Olearnik B, Boratyński J. Removal of endotoxins from bacteriophage preparations by extraction with organic solvents. PLoS ONE, 2015, 10(3): e0122672.

[39] Türkay S, Civelekoğlu H. Deacidification of sulfur olive oil. II. Multi-stage liquid-liquid extraction of miscella with ethyl alcohol. J Am Oil Chem Soc, 1991, 68(11): 818-821.

[40] Unrean P, Nguyen NH, Visessanguan W, Kitsubun P. Improvement of nattokinase production by Bacillus subtilis using an optimal feed strategy in fed-batch fermentation. KKU Res J, 2012, 17: 769-777.

[41] Wu HS, Wang YJ. Salting-out effect on recovery of 1, 3-propanediol from fermentation broth. Ind Eng Chem Res, 2012, 51(33): 10930-10935.

[42] Xiao S, Ju L-K. Energy-efficient ultrasonic release of bacteria and particulates to facilitate ingestion by phagotrophic algae for waste sludge treatment and algal biomass and lipid production. Chemosphere, 2018, 209: 588-598.

[43] Zhang G, Ma B, Xu X, Li C, Wang L. Fast conversion of glycerol to 1, 3-propanediol by a new strain of Klebsiella pneumoniae. Biochem Eng J, 2007, 37(3): 256-260.

[44] Zhang ZR, Shen JT, Dai JY, Sun YQ, Dong YS, Xiu ZL. Separation and purification of Klebsiella phage by two-step salting-out extraction. Sep Purif Technol, 2020, 242: 116784.

[45] Zhang C, Sharma S, Wang W, Zeng AP. A novel downstream process for highly pure 1,3-propanediol from an efficient fed-batch fermentation of raw glycerol by Clostridium pasteurianum. Eng Life Sci, 2021, 21(6): 351-363.

[46] Zheng K, Jiang L, Yu S, Xian M, Song Z, Liu S, Xu C. The design and synthesis of high efficiency adsorption materials for 1, 3-propanediol: physical and chemical structure regulation. RSC Adv, 2020, 10(62): 38085-38096.

[47] Zurabov F, Zhilenkov E. Characterization of four virulent Klebsiella pneumoniae bacteriophages, and evaluation of their potential use in complex phage preparation. Virol J, 2021, 18(1): 1-20.

Funding

National Natural Science Foundation of China(22078042)