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Enhanced production of ε-poly-l-lysine by immobilized Streptomyces ahygroscopicus through repeated-batch or fed-batch fermentation with in situ product removal

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Abstract

ε-Poly-l-lysine (ε-PL) is a naturally-occurring l-lysine homopolymer having a broad-spectrum antimicrobial activity and used widely as a food preservative. In the present study, the combined use of immobilization and in situ product removal (ISPR) was evaluated for the production of ε-PL by Streptomyces ahygroscopicus GIM8. Results showed that ε-PL production in the flask cultures decreased from 0.84 to 0.38–0.56 g/L upon immobilization on loofah sponge with different amounts (0.5–3 g in 50 mL medium in a flask). By applying continuous ISPR to the immobilized flask cultures, ε-PL production as high as 3.51 g/L was obtained compared to 0.51 g/L of the control. A satisfactory titer of 1.84 g/L ε-PL could also be achieved with intermittent ISRP (three cycles of ISPR operation during cultivation). Further investigation showed that low levels of ε-PL retained in the broth appeared to favor its biosynthesis. In the repeated-batch fermentation in a 5 L immobilized bioreactor, with continuous ISPR, the final average ε-PL concentration and productivity were 3.35 g/L and 0.797 g/L/day, respectively, and 3.18 g/L and 0.756 g/L/day for the alternative (intermittent ISPR), in comparison to 1.16 g/L and 0.277 g/L/day with no ISPR usage. In the fed-batch fermentation with immobilized cells, the combined use of intermittent ISPR and extra nutrient feeding increased ε-PL concentration and productivity up to 24.57 g/L and 9.34 g/L/day. The fermentation processes developed could serve as an effective approach for ε-PL production and, moreover, the combination could greatly simplify downstream processing for ε-PL separation and purification.

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References

  1. Shima S, Sakai H (1977) Polylysine produced by Streptomyces. Agric Biol Chem 41:1807–1809

    CAS  Google Scholar 

  2. Hiraki J (2000) Basic and applied studies on ε-polylysine. J Antibact Antifungal Agents 23:349–354

    Google Scholar 

  3. Shima S, Fukuhara Y, Sakai H (1982) Inactivation of bacteriophages by ε-poly-l-lysine produced by Streptomyces. Agric Biol Chem 46:1917–1919

    CAS  Google Scholar 

  4. Shima S, Matsuoka H, Iwamoto T, Sakai H (1984) Antimicrobial action of ε-poly-l-lysine. J Antibiot 37:1449–1455

    Article  CAS  Google Scholar 

  5. Pandey AK, Kumar A (2014) Improved microbial biosynthesis strategies and multifarious applications of the natural biopolymer epsilon-poly-l-lysine. Process Biochem 49:496–505

    Article  CAS  Google Scholar 

  6. Hiraki J, Ichikawa T, Ninomiya S, Seki H, Uohama K, Seki H, Kimura S, Yanagimoto Y, Barnett JW (2003) Use of ADME studies to confirm the safety of ε-polylysine as a preservative in food. Regul Toxicol Pharmacol 37:328–340

    Article  CAS  PubMed  Google Scholar 

  7. Shih IL, Shen MH, Van YT (2006) Microbial synthesis of poly(ε-lysine) and its various applications. Bioresour Technol 97:1148–1159

    Article  CAS  PubMed  Google Scholar 

  8. Hamano Y, Nicchu I, Shimizu T, Onji Y, Hiraki J, Takagi H (2007) ε-Poly-l-lysine producer, Streptomyces albulus, has feedback-inhibition resistant aspartokinase. Appl Microbiol Biotechnol 76:873–882

    Article  CAS  PubMed  Google Scholar 

  9. Shima S, Oshima S, Sakai H (1983) Biosynthesis of ε-poly-l-lysine by washed mycelium of Streptomyces albulus No. 346. Nippon Nogeikagaku Kaishi 57:221–226

    Article  CAS  Google Scholar 

  10. Chen XS, Ren XD, Zeng X, Zhao FL, Tang L, Zhang HJ, Mao ZG (2013) Enhancement o f ε-poly-l-lysine production coupled with precursor l-lysine feeding in glucose-glycerol co-fermentation by Streptomyces sp. M-Z18. Bioprocess Biosyst Eng 36:1843–1849

    Article  CAS  PubMed  Google Scholar 

  11. Yamanaka K, Maruyama C, Takagi H, Hamano Y (2008) ɛ-Poly-l-lysine dispersity is controlled by a highly unusual nonribosomal peptide synthetase. Nat Chem Biol 4:766–772

    Article  CAS  PubMed  Google Scholar 

  12. Shukla SC, Singh A, Pandey AK, Mishra A (2012) Review on production and medical applications of ɛ-poly-l-lysine. Biochem Eng J 65:70–81

    Article  CAS  Google Scholar 

  13. Wang L, Chen XS, Wu GY, Li S, Zeng X, Ren XD, Tang L, Mao ZG (2017) Enhanced ε-poly-l-lysine production by inducing double antibiotic-resistant mutations in Streptomyces albulus. Bioprocess Biosyst Eng 40:271–283

    Article  CAS  PubMed  Google Scholar 

  14. Chen XS, Tang L, Li S, Liao LJ, Zhang JH, Mao ZG (2011) Optimization of medium for enhancement of ε-poly-l-lysine production by Streptomyces sp. M-Z18 with glycerol as carbon source. Bioresour Technol 102:1727–1732

    Article  CAS  PubMed  Google Scholar 

  15. Bankar SB, Singhal RS (2011) Improved poly-ε-lysine biosynthesis using Streptomyces noursei NRRL 5126 by controlling dissolved oxygen during fermentation. J Microbiol Biotechnol 21:652–658

    Article  CAS  PubMed  Google Scholar 

  16. Chen XS, Li S, Liao LJ, Ren XD, Li F, Tang L, Zhang JH, Mao ZG (2011) Production of ε-poly-l-lysine using a novel two-stage pH control strategy by Streptomyces sp. M-Z18 from glycerol. Bioprocess Biosyst Eng 34:561–567

    Article  CAS  PubMed  Google Scholar 

  17. Ren XD, Chen XS, Zeng X, Wang L, Tang L, Mao ZG (2015) Acidic pH shock induced overproduction of ε-poly-l-lysine in fed-batch fermentation by Streptomyces sp. M-Z18 from agro-industrial by-products. Bioprocess Biosyst Eng 38:1113–1125

    Article  CAS  PubMed  Google Scholar 

  18. Hirohara H, Takehara M, Saimura M, Masayuki A, Miyamoto M (2006) Biosynthesis of poly(ε-l-lysine)s in two newly isolated strains of Streptomyces sp. Appl Microbiol Biotechnol 73:321–331

    Article  CAS  PubMed  Google Scholar 

  19. Kahar P, Iwata T, Hiraki J, Park E, Okabe M (2001) Enhancement of ε-polylysine production by Streptomyces albulus strain 410 using pH control. J Biosci Bioeng 91:190–194

    Article  CAS  PubMed  Google Scholar 

  20. Kito M, Takimoto R, Yoshida T, Nagasawa T (2002) Purification and characterization of an ε-poly-l-lysine-degrading enzyme from an ε-poly-l-lysine-producing strain of Streptomyces albulus. Arch Microbiol 178:325–330

    Article  CAS  PubMed  Google Scholar 

  21. Liu Y, Liu D (2004) Kinetic study on glycerol production by repeated batch fermentation using free Candida krusei. Process Biochem 39:1507–1510

    Article  CAS  Google Scholar 

  22. Chen CC, Lan CC, Pan CL, Huang MY, Chew CH, Hung CC, Chen PH, Victor Lin HT (2019) Repeated-batch lactic acid fermentation using a novel bacterial immobilization technique based on a microtube array membrane. Process Biochem 87:25–32

    Article  CAS  Google Scholar 

  23. Suwannakham S, Yang ST (2005) Enhanced propionic acid fermentation by Propionibacterium acidipropionici mutant obtained by adaptation in a fibrous-bed bioreactor. Biotechnol Bioeng 91:325–337

    Article  CAS  PubMed  Google Scholar 

  24. Huang L, Lacroix C, Daba H, Simard RE (1996) Pediocin 5 production and plasmid stability during continuous free and immobilized cell cultures of Pediococcus acidilactici UL5. J Appl Bacteriol 80:635–644

    Article  CAS  PubMed  Google Scholar 

  25. Yang XH, Wang BW, Cui FN, Tan TW (2005) Production of lipase by repeated batch fermentation with immobilized Rhizopus arrhizus. Process Biochem 40:2095–2103

    Article  CAS  Google Scholar 

  26. Dishisha T, Alvarez MT, Hatti-Kaul R (2012) Batch- and continuous propionic acid production from glycerol using free and immobilized cells of Propionibacterium acidipropionici. Bioresour Technol 118:553–562

    Article  CAS  PubMed  Google Scholar 

  27. Meleigy SA, Khalaf MA (2009) Biosynthesis of gibberellic acid from milk permeate in repeated batch operation by a mutant Fusarium moniliforme cells immobilized on loofa sponge. Bioresour Technol 100:374–379

    Article  CAS  PubMed  Google Scholar 

  28. Zhang Y, Feng XH, Xu H, Yao Z, Ouyang PK (2010) ε-Poly-l-lysine production by immobilized cells of Kitasatospora sp. MY 5–36 in repeated fed-batch cultures. Bioresour Technol 101:5523–5527

    Article  CAS  PubMed  Google Scholar 

  29. Stark D, von Stochar U (2003) In situ product removal (ISPR) in whole biotechnology during the last twenty years. Adv Biochem Eng Biotechnol 80:149–175

    CAS  PubMed  Google Scholar 

  30. Pongtharangku T, Demirci A (2007) Online recovery of nisin during fermentation and its effect on nisin production in biofilm reactor. Appl Microbiol Biotechnol 74:555–562

    Article  CAS  PubMed  Google Scholar 

  31. Singhvi M, Zendo T, Gokhale D, Sonomoto K (2018) Greener l-lactic acid production through in situ extractive fermentation by an acid-tolerant Lactobacillus strain. Appl Microbiol Biotechnol 102:6425–6435

    Article  CAS  PubMed  Google Scholar 

  32. Eiki H, Gushima H, Saito T, Ishida H, Oka Y, Osono T (1988) Product inhibition and its removal on josamycin fermentation by Streptomyces narbonensis var. josamyceticus. J Biosci Bioeng 66:559–565

    CAS  Google Scholar 

  33. Zhang LJ, Jin ZH, Chen XG, Jin QC, Feng MG (2012) Glycine feeding improves pristinamycin production during fermentation including resin for in situ separation. Bioprocess Biosyst Eng 35:513–517

    Article  CAS  PubMed  Google Scholar 

  34. Liu SR, Wu QP, Zhang JM, Mo SP (2011) Production of ε-poly-l-lysine by Streptomyces sp. using resin-based, in situ product removal. Biotechnol Lett 33:1581–1585

    Article  CAS  PubMed  Google Scholar 

  35. Huang JM, Wu QP, Liu SR, Zhang JM (2011) Screening of new ε-polylysine producing strain and structure identification of its product. Microbiol China 38:871–877 (in Chinese)

    CAS  Google Scholar 

  36. Wang P, He JY, Yin JF (2015) Enhanced biocatalytic production of l-cysteine by Pseudomonas sp. B-3 with in situ product removal using ion-exchange resin. Bioprocess Biosyst Eng 38:421–428

    Article  CAS  PubMed  Google Scholar 

  37. Itzhaki FR (1972) Colorimetric method for estimating polylysine and polyarginine. Anal Biochem 50:569–574

    Article  CAS  PubMed  Google Scholar 

  38. Kar S, Swain MR, Ray RC (2009) Statistical optimization of alpha-amylase production with immobilized cells of Streptomyces erumpens MTCC 7317 in Luffa cylindrical L. sponge discs. Appl Biochem Biotechnol 152:177–188

    Article  CAS  PubMed  Google Scholar 

  39. Saudagar PS, Shaligram NS, Singhal RS (2008) Immobilization of Streptomyces clavuligerus on loofah sponge for the production of clavulanic acid. Bioresour Technol 99:2250–2253

    Article  CAS  PubMed  Google Scholar 

  40. Cao NJ, Du JX, Gong CS, Tsao GT (1996) Simultaneous production and recovery of fumaric acid from immobilized Rhizopus oryzae with a rotary biofilm contactor and an adsorption column. Appl Environ Microbiol 62:2926–2931

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Roddick FA, Britz ML (1997) Production of hexanoic acid by free and immobilized cells of Megasphaera elsdenii: influence of in-situ product removal using ion exchange resin. J Chem Tech Biotechnol 69:383–391

    Article  CAS  Google Scholar 

  42. Xue C, Zhao JB, Lu CC, Yang ST, Bai FW, Tang IC (2012) High-titer n-butanol production by Clostridium acetobutylicum JB200 in fed-batch fermentation with intermittent gas stripping. Biotechnol Bioeng 109:2746–2756

    Article  CAS  PubMed  Google Scholar 

  43. Liu J, Guo T, Luo Y, Chai X, Wu J, Zhao W, Jiao P, Luo F, Lin Q (2019) Enhancement of Monascus pigment productivity via a simultaneous fermentation process and separation system using immobilized-cell fermentation. Bioresour Technol 272:552–560

    Article  CAS  PubMed  Google Scholar 

  44. Liu SR, Wu QP, Zhang JM, Mo SP, Yang XJ, Xiao C (2012) Enhanced ε-poly-l-lysine production from Streptomyces ahygroscopicus by a combination of cell immobilization and in situ adsorption. J Microbiol Biotechnol 22:1218–1223

    Article  CAS  PubMed  Google Scholar 

  45. Li Q, Wang D, Hu G, Xing J, Su Z (2011) Integrated bioprocess for high-efficiency production of succinic acid in an expanded-bed adsorption system. Biochem Eng J 56:150–157

    Article  CAS  Google Scholar 

  46. Yang XP, Tsao GT (1995) Enhanced acetone-butanol fermentation using repeated fed-batch operation coupled with cell recycle by membrane and simultaneous removal of inhibitory products by adsorption. Biotechnol Bioeng 47:444–450

    Article  CAS  PubMed  Google Scholar 

  47. Kahar P, Kobayashi K, Iwata T, Hiraki J, Kojima M, Okabe M (2002) Production of ɛ-polylysine in an airlift bioreactor (ABR). J Biosci Bioeng 93:274–280

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

This work was financially supported by the Key Cultivating Program of Ningde Normal University (2018ZDK01) and the Industry-Leading Program of the Science and Technology Bureau of Fujian Province (2015N0032).

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Correspondence to Sheng-Rong Liu.

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Liu, SR., Yang, XJ. & Sun, DF. Enhanced production of ε-poly-l-lysine by immobilized Streptomyces ahygroscopicus through repeated-batch or fed-batch fermentation with in situ product removal. Bioprocess Biosyst Eng 44, 2109–2120 (2021). https://doi.org/10.1007/s00449-021-02587-7

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