Abstract
It has been half a century since investigators first began experimenting with adding ion exchange resins during the fermentation of microbial natural products. With the development of nonionic polymeric adsorbents in the 1970s, the application of in situ product adsorption in bioprocessing has grown slowly, but steadily. To date, in situ product adsorption strategies have been used in biotransformations, plant cell culture, the production of biofuels, and selected bulk chemicals, such as butanol and lactic acid, as well as in more traditional natural product fermentation within the pharmaceutical industry. Apart from the operational gains in efficiency from the integration of fermentation and primary recovery, the addition of adsorbents during fermentation has repeatedly demonstrated the capacity to significantly increase titers by sequestering the product and preventing or mitigating degradation, feedback inhibition and/or cytotoxic effects. Adoption of in situ product adsorption has been particularly valuable in the early stages of natural product-based drug discovery programs, where quickly and cost-effectively generating multigram quantities of a lead compound can be challenging when using a wild-type strain and fermentation conditions that have not been optimized. While much of the literature involving in situ adsorption describes its application early in the drug development process, this does not imply that the potential for scale-up is limited. To date, commercial-scale processes utilizing in situ product adsorption have reached batch sizes of at least 30,000 l. Here we present examples where in situ product adsorption has been used to improve product titers or alter the ratios among biosynthetically related natural products, examine some of the relevant variables to consider, and discuss the mechanisms by which in situ adsorption may impact the biosynthesis of microbial natural products.
Similar content being viewed by others
References
Abrams IM, Millar JR (1997) A history of the origin and development of macroporous ion-exchange resins. React Funct Polym 35:7–22
Arslanian RL, Parker CD, Wang PK, McIntire JR, Lau J, Starks C, Licari PJ (2002) Large-scale isolation and crystallization of epothilone D from Myxococcus xanthus cultures. J Nat Prod 65:570–572
Bechtold M, Panke S (2009) In situ product recovery integrated with biotransformations. Chemia 63(6):345–348
Benigni D, Gougoutas JZ, DiMarco JD (2006) Method for the preparation, isolation and purification of epothilone B and x-ray crystal structures of epothilone B. US Patent Application No. US 2006/0135474 A1
Bo J, Jin ZH, Lei YL, Mei LH, Li NH (2006) Improved production of pristinamycin coupled with the adsorbent resin in fermentation by Streptomyces pristinaespiralis. Biotechnol Lett 28:1811–1815
Boot CM, Gassner NC, Compton JE, Tenney K, Tramble CM, Lokey SR, Holman TR, Crews P (2007) Pinpointing pseurotins from marine-derived Aspergillus as tools for chemical genetics using synthetic lethality yeast screen. J Nat Prod 70:1672–1675
Carlson JC, Shengying Li, Burr DA, Sherman DH (2009) Isolation and characterization of Tirandamycins from a marine-derived Streptomyces sp. J Nat Prod 72:2076–2079
Cella R, Vining LC (1975) Resistance to streptomycin in a producing strain of Streptomyces griseus. Can J Microbiol 21:463–472
Cundliffe E, Demain AL (2010) Avoidance of suicide in antibiotic-producing microbes. J Ind Microbiol Biotechnol 37:643–672
Denkewalter RG, Gillin J (1959) Verfahren zur Gewinnung von antibiotica. Auslegreshrift 1062891
Desai MC, Chackalamannil S (2008) Rediscovering the role of natural products in drug discovery. Curr Opin Drug Discov Dev 11(4):436–437
Fierro JF, Hardisson C, Salas JA (1988) Involvement of cell impermeability in resistance to macrolides in some producer streptomycetes. J Antibiot 41(1):142–144
Fontanals N, Marcé RM, Borrull F (2005) New hydrophilic materials for solid-phase extraction. Trends Anal Chem 24(5):394–406
Freeman A, Woodley JM, Lilly MD (1993) In situ product removal as a tool for bioprocessing. Biotechnology 11:1007–1012
Frykman S, Tsuruta H, Galazzo J, Licari P (2006) Characterization of product capture resin during microbial cultivation. J Ind Microbiol Biotechnol 33:445–453
Frykman SA, Tsuruta H, Licari PJ (2005) Assessment of fed-batch, semicontinuous and continuous epothilone D production processes. Biotechnol Prog 21:1102–1108
Fujii N, Katsuyama T, Kobayashi K, Hara M, Nakano H (1995) The clecarmycins, new antitumor antibiotic produced by streptomyces: fermentation, isolation and biological properties. J Antibiot 48(8):768–772
Gastaldo L, Marinelli f, Acquarella C, Restelli E, Quarta C (1996) Improvement of the kirromycin fermentation by resin addition. J Ind Microbiol 16:305–308
Gerth K, Bedorf N, Irschik H, Höfle G (1994) The soraphens: a family of novel antifungal compounds from Sorangium cellulosum (Myxobacteria). J Antibiot 47(1):23–31
Gerth K, Pradella S, Perlova O, Beyer S, Müller R (2003) Myxobacteria: proficient producers of novel natural products with various biological activities—past and future biotechnological aspects with the focus on the genus Sorangium. J Biotechnol 106:233–253
Gerth K, Washausen P, Höfle G, Irschik H, Reichenback H (1995) The jerangolids: a family of new antifungal compounds from Sorangium cellulosum (Myxobacteria). J Antibiot 49(1):71–75
Hara M, Asano K, Kawamoto I, Takiguchi T, Katsumata S, Takahashi KI, Nakano H (1989) Leinamycin, a new antitumor antibiotic from Streptomyces, producing organism, fermentation and isolation. J Antibiot 42(12):1768–1774
Hara M, Takiguchi T, Ashizawa T, Gomi K, Nakano H (1991) Sapurimycin, new antitumor antibiotic produced by Streptomyces. Producing organism, fermentation, isolation and biological properties. J Antibiot 44(1):33–39
Harvey AL (2008) Natural products in drug discovery. Drug Discov Today 13(19/20):894–901
He H, Ding WD, Berman VS, Richardson AD, Ireland CM, Greenstein M, Ellestad GA, Carter GT (2001) Lomaiviticins A and B, potent antitumor antibiotics from Micromonospora lomaivitienses. J Am Chem Soc 123:5362–5363
Hollmann D, Merrettig-Bruns U, Müller U, Onken U (1990) Secondary metabolites by extractive fermentation, Separations for biotechnology 2. Papers presented at the 2nd international symposium on separations for biotechnology 2, 567–576
Inoue K, Yamazaki M, Armentrout RW (1991) Process for producing streptovaricin. European Patent Application 91309793.7, EP 0 482 908 A2
Jarvis BB, Armstrong CA, Zeng M (1990) Use of resin for trichothecene production in liquid cultures. J Antibiotic 43(11):1502–1504
Junker BH (2004) Scale-up methodologies for Escherichia coli and yeast fermentation processes. J Biosci Bioeng 97(6):347–364
Karwowski JP, Jackson M, Sunga G, Sheldon P, Poddig JB, Kohl WL, Kadam S (1994) Dorrigocins: novel antifungal antibiotics that change the morphology of ras-transformed NIH/3T3 cells to that of normal cells. J Antibiot 47(8):862–869
Kim BH, Andersen C, Benz R (2001) Identification of a cell wall channel of Streptomyces griseus: the channel contains a binding site for streptomycin. Mol Microbiol 41(3):655–673
Kim CH, Kim SW, Hong SI (2000) An integrated fermentation-separation process for the production of red pigment by Serratia sp. KH-95. Process Biochem 35:485–490
Kim JJ, Lim SK, Lee MO, Lim SM, Lee B, Kim DH (2009) Method of extraction and yield-up of tricyclo compounds by adding a solid adsorbent as their carriers in fermentation medium. WO 2009/025439 A1
Knight V, Sanglier JJ, DiTullio D, Braccili S, Bonner P, Waters J, Hughes D, Zhang L (2003) Diversifying microbial natural products for drug discovery. Appl Microbiol Biotechnol 62:446–458
Kusunose Y, Wang DIC (2004) Enhancement of production of phenylalanine using uncharged polymeric beads. Chem Eng Comm 191:1199–1207
Kusunose Y, Wang DIC (2004) Preliminary studies on extractive fermentation of phenylalanine using uncharged polymeric beads. Chem Eng Comm 191:1185–1198
Lam KS, Gustavson DR, Veitch JA, Forenza S (1993) The effect of cerulenin on the production of esperamicin A1 by Actinomadura verrucosospora. J Ind Microbiol 12:99–102
Lam KS, Veitch JA, Lowe SE, Forenza S (1995) Effect of neutral resins on the production of dynemicins by Micromonospora chersina. J Ind Microbiol 15:453–456
Lau J, Frykman SA, Regentin R, Ou S, Tsuruta H, Licari PJ (2002) Optimizing the heterologous production of Epothilone D in Myxococcus xanthus. Biotechnol Bioeng 78(3):280–288
Leaf TA, Desai RP, Licari P, Woo EJ (2005) Method of producing a compound by fermentation. US 2005/0130283 A1
Lee JC, Min JW, Park DJ, Son KH, Yoon KH, Park HR, Park YS, Kwon MG, Lee JM, Kim CJ (2005) Large-scale fermentation for the production of teicoplanin from a mutant of Actinoplanes teicomyceticus. J Microbiol Biotechnol 15(4):787–791
Lee JC, Park HR, Park DJ, Lee HB, Kim YB, Kim CJ (2003) Improved production of teicoplanin using adsorbent resin in fermentation. Lett Appl Microbiol 37:196–200
Liu B, Hui J, Cheng YQ, Zhang X (2012) Extractive fermentation for enhanced production of thailandepsin A from Burkhoderia thailandensis E264 using polyaromatic adsorbent resin Diaion HP20. J Ind Microbiol Biotechnol 39:767–776
Magarvey NA, Keller JM, Dernan V, Dworkin M, Sherman DH (2004) Isolation and characterization of novel marine-derived actinomycetes taxa rich in bioactive metabolites. Appl Environ Microbiol 70:7520–7529
Malla S, Niraula NP, Liou K, Sohng JK (2009) Self-resistance mechanism in Streptomyces peucetius: overexpression odrrA, drrB and drrC for doxorubicin enhancement. Microb Res 165:259–267
Marshall VP, McWethy JS, Visser J, Cialdella JI, Laborde AL (1987) Current fermentation technology from actinomycetes: the example of paulomycin. Dev Ind Microbiol 28:105–114 J Ind Microbiol. Suppl No. 2
Marshall VP, McWethy SJ, Sirotti JM, Cialdella JI (1990) The effect of neutral resins on the fermentation production of rubradirin. J Ind Microbiol 5:283–288
Martin JF, Casqueiro J, Liras P (2005) Secretion systems for secondary metabolites: how producer cells sent out messages of intercellular communication. Curr Opin Microbiol 8:282–293
McDonald LA, Abbanat DR, Barbieri LR, Bernan VS, Discafani CM, Greenstein M, Janota D, Korshalla JD, Lassota P, Tischler M, Carter GT (1999) Spiroxins, DNA cleaving antitumor antibiotics from a marine-derived fungus. Tetrahedron Lett 40:2489–2492
Millitzer M, Wenzig E, Peukert W (2005) Process modeling of in situ adsorption of a bacterial lipase. Biotech Bioeng 92(6):789–801
Monteagudo JM, Aldavero M (1999) Production of L-lactic acid by Lactobacillus delbrueckii in chemostat culture using an ion exchange resin system. J Chem Technol Biotechnol 74:627–634
Nagata, H Ochiai K, Aotani Y, Ando K, Yoshida M, Takahashi I, Tamaoki T (1997) Lymphostin (LK6-A), a novel immunosuppressant from Streptomyces sp. KY11783: taxonomy of the producing organism, fermentation, Isolation and biological activities. J Antibiot 50(7):537–540
Newman DJ, Cragg GM (2007) Natural products as a source of new drugs over the last twenty five years. J Nat Prod 70:461–477
Park SW, Han SE, Kim DS, Sim SJ (2007) Improvement of epothilone B production by in situ removal of ammonium using cation exchange resin in Sorangium cellulosum culture. Biochem Eng Journal 37:328–331
Payne GF, Wang HY (1989) The effect of feedback regulation and in situ product removal on the conversion of sugar to cyclohexamide by Streptomyces griseus. Arch Microbiol 151:331–335
Potterat O (1994) Zähner (1993) Exfoliamycin and related metabolites, new naphthoquinone antibiotics from Streptomyces exfoliates. J Antibiot 46(2):346–349
Qureshi N, Hughes S, Maddox IS, Cotta MA (2005) Energy–efficient recovery of butanol from model solutions and fermentation broth by adsorption. Bioprocess Biosyst Eng 27:215–222
Robins RJ, Rhodes MJC (1986) The stimulation of anthraquinone production by Cinchona lidgeriana cultures with polymeric adsorbents. Appl Microbiol Biotechnol 24:35–41
Rokem SJ, Hurley LH (1981) Sensitivity and permeability of the antramycin producing organism Streptomyces refuineus to anthrmycin and structurally related antibiotics. J Antibiot 34(9):1171–1174
Shin CS, Ahn BW, Lee SH, Sung UK, Bok SH (1988) Liberation of sisomicin from cells by sodium chloride. Appl Microbiol Biotechnol 28:37–38
Shue YK, Du F, Chiou, MH, Wu MC, Chen YT, Duffield J, Okumu FW (2004) Tiacumicin Production. WO 2004/014295 A2
Sim SJ, Chang HN (1997) Shikonin production by hairy roots of Lithospermum erythrorhizon in bioreactors with in situ separation. Hairy Roots 1997:219–225
Singh MP, Leighton MM, Barbieri LR, Roll DM, Urbance SE, Hoshan L, McDonald LA (2010) Fermentative production of self toxic fungal secondary metabolites. J Ind Microbiol Biotechnol 37:335–340
Stark D, von Stockar U (2003) In-situ product removal (ISPR) in whole cell biotechnology during the last twenty years. Adv Biochem Eng Biotechnol 80:149–175
Sugiyama M, Mizuno S, Ohta Y, Mochizuki H, Nimi O (1990) Kinetic studies of streptomycin uptake implicated in self-resistance in streptomycin producer. Biotechnol Lett 12(1):1–6
Tsueng G, Lam KS (2007) Stabilization effect of resin on the production of potent proteasome inhibitor NPI-0052 during submerged fermentation of Salinispora tropica. J Antibiot 60(7):469–472
Viloria-Cols ME, Hatti-Kaul R, Mattiasson B (2004) Agarose-coated anion exchanger prevents cell-adsorbent interactions. J Chromatogr A 1043:195–200
Voelker F, Altaba S (2001) Nitrogen source governs the patterns of growth and pristinamycin production in Streptomyces pristinaespiralis. Microbiol 147:2447–2459
Wang HY, Kominek LA, Jost JL (1981) On-line extraction of fermentation processes. In: Moo Young M, Robinson CW, Vezina C (eds) Advances in Biotechnology I. Scientific and engineering principles. Pergamon Press, Oxford, pp 601–607
Warr GA, Veitch JA, Walsh AW, Hesler GA, Pirnik DM, Leet JE, Lin PFM, Medina IA, McBrien KD, Forenza A, Clark JM, Lam KS (1996) BMS-182123, a fungal metabolite that inhibits the production of TNF-α by macrophages and monocytes. J Antibiot 49(3):234–240
Williams RD, Chauret N, Bedard C, Archambault J (1992) Effect of polymeric adsorption of sanguinarine by Papaver somniferum cell cultures. Biotech Bioeng 40(8):971–977
Woo EJ, Starks CM, Carney JR, Arslanian R, Cadapan L, Zavala S, Licari P (2002) Migrastatin and a new compound, Isomigrastatin, from Streptomyces platensis. J Antibiot 55(2):141–146
Xu LJ, Liu YS, Zhou LG, Wu JY (2009) Enhanced beauvericin production with in situ adsorption in mycelial liquid culture of Fusarium redolens Dzf2. Process Biochem 44:1063–1067
Yamazaki M, Onoue K (1993) Process for the preparation of streptovaricin by fermentation. European Patent Application No. 92311235.3, EP 0 546 819 A1
Yu PL, Dunn NW, Kim WS (2002) Lactate removal by anionic-exchange resin improves nisin production by Lactococcus lactis. Biotechnol Lett 24:59–64
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Phillips, T., Chase, M., Wagner, S. et al. Use of in situ solid-phase adsorption in microbial natural product fermentation development. J Ind Microbiol Biotechnol 40, 411–425 (2013). https://doi.org/10.1007/s10295-013-1247-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10295-013-1247-9