Abstract
In Gluconobacter oxydans cultivations on glucose, CaCO3 is typically used as pH-buffer. This buffer, however, has disadvantages: suspended CaCO3 particles make the medium turbid, thereby, obstructing analysis of microbial growth via optical density and scattered light. Upon searching for alternative soluble pH-buffers, bacterial growth and productivity was inhibited most probably due to osmotic stress. Thus, this study investigates in detail the osmotic sensitivity of G. oxydans ATCC 621H and DSM 3504 using the Respiratory Activity MOnitoring System. The tested soluble pH-buffers and other salts attained osmolalities of 0.32–1.19 osmol kg−1. This study shows that G. oxydans ATCC 621H and DSM 3504 respond quite sensitively to increased osmolality in comparison to other microbial strains of industrial interest. Osmolality values of >0.5 osmol kg−1 should not be exceeded to avoid inhibition of growth and product formation. This osmolality threshold needs to be considered when working with soluble pH-buffers.
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References
Ameyama M, Matsushita K, Ohno Y, Shinagawa E, Adachi O (1981) Existence of a novel prosthetic group, PQQ, in membrane-bound, electron transport chain-linked, primary dehydrogenases of oxidative bacteria. FEBS Lett 130(2):179–183. doi:10.1016/0014-5793(81)81114-3
Anderlei T, Büchs J (2001) Device for sterile online measurement of the oxygen transfer rate in shaking flasks. Biochem Eng J 7(2):157–162. doi:10.1016/S1369-703X(00)00116-9
Anderlei T, Mrotzek C, Bartsch S, Amoabediny G, Peter CP, Büchs J (2007) New method to determine the mass transfer resistance of sterile closures for shaken bioreactors. Biotechnol Bioeng 98(5):999–1007. doi:10.1002/bit.21490
Anderlei T, Zang W, Papaspyrou M, Büchs J (2004) Online respiration activity measurement (OTR, CTR, RQ) in shake flasks. Biochem Eng J 17(3):187–194. doi:10.1016/S1369-703x(03)00181-5
Asai T (1968) Acetic Acid Bacteria: Classification and Biochemical Activities. University of Tokyo Press, Tokyo
Beschkov V, Velizarov S, Peeva L (1995) Some kinetic aspects and modeling of biotransformation of d-glucose to keto-d-gluconates. Bioprocess Eng 13:301–305. doi:10.1007/BF00369561
Brown AD (1990) Microbial water stress physiology: principles and perspectives. Wiley, Chichester
Brown DA, Cook RA (1981) Role of metal cofactors in enzyme regulation. Differences in the regulatory properties of the Escherichia coli nicotinamide adenine dinucleotide phosphate specific malic enzyme, depending on whether magnesium ion or manganese ion serves as divalent cation. Biochemistry 20(9):2503–2512. doi:10.1021/bi00512a022
Buchenauer A, Hofmann MC, Funke M, Büchs J, Mokwa W, Schnakenberg U (2009) Micro-bioreactors for fed-batch fermentations with integrated online monitoring and microfluidic devices. Biosens Bioelectron 24(5):1411–1416. doi:10.1016/j.bios.2008.08.043
Davey KR (1989) A predictive model for combined temperature and water activity on microbial-growth during the growth-phase. J Appl Bacteriol 67(5):483–488. doi:10.1111/j.1365-2672.1989.tb02519.x
Davey KR (1991) Applicability of the Davey (Linear Arrhenius) predictive model to the lag phase of microbial-growth. J Appl Bacteriol 70(3):253–257. doi:10.1111/j.1365-2672.1991.tb02933.x
Duggan PF (1977) An enzyme system requiring magnesium, calcium and potassium ions. Proc R Ir Acad B 77(19–47):449–455
Ellis KJ, Morrison JF (1982) Buffers of constant ionic strength for studying pH-dependent processes. Methods Enzymol 87:405–426. doi:10.1016/S0076-6879(82)87025-0
Funke M, Buchenauer A, Schnakenberg U, Mokwa W, Diederichs S, Mertens A, Müller C, Kensy F, Büchs J (2010) Microfluidic BioLector-Microfluidic bioprocess control in microtiter plates. Biotechnol Bioeng 107(3):497–505. doi:10.1002/Bit.22825
Gao L, Hu Y, Liu J, Du G, Zhou J, Chen J (2014) Stepwise metabolic engineering of Gluconobacter oxydans WSH-003 for the direct production of 2-keto-l-gulonic acid from d-sorbitol. Metab Eng 24:30–37. doi:10.1016/j.ymben.2014.04.003
Good NE, Izawa S (1972) Hydrogen ion buffers. Methods Enzymol 24:53–68. doi:10.1016/0076-6879(72)24054-X
Greenfield S, Claus G (1972) Nonfunctional tricarboxylic acid cycle and the mechanism of glutamate biosynthesis in Acetobacter suboxydans. J Bacteriol 112(3):1295–1301
Griffin DM (1981) Water and micobial stress. Adv Microb Ecol 5:91–136. doi:10.1007/978-1-4615-8306-6_3
Guillouet S, Engasser JM (1995) Sodium and proline accumulation in Corynebacterium glutamicum as a response to an osmotic saline upshock. Appl Microbiol Biotechnol 43(2):315–320. doi:10.1007/BF00172831
Gupta A, Singh VK, Qazi GN, Kumar A (2001) Gluconobacter oxydans: its biotechnological applications. J Mol Microbiol Biotechnol 3(3):445–456
Hommel R, Ahnert P (2000) Gluconobacter. Encyclopedia of food microbiology. Academic Press, London
Jeude M, Dittrich B, Niederschulte H, Anderlei T, Knocke C, Klee D, Büchs J (2006) Fed-batch mode in shake flasks by slow-release technique. Biotechnol Bioeng 95:433–445. doi:10.1002/bit.21012
Kandegedara A, Rorabacher DB (1999) Noncomplexing tertiary amines as “better” buffers covering the range of pH 3-11. Temperature dependence of their acid dissociation constants. Anal Chem 71(15):3140–3144. doi:10.1021/ac9902594
Kumar S, Wittmann C, Heinzle E (2004) Review: minibioreactors. Biotechnol Lett 26(1):1–10. doi:10.1023/b:bile.0000009469.69116.03
Lichtenthaler FW (2006) The key sugars of biomass: Availability, present non-food applications and potential industrial development lines. Biorefineries, biobased industrial processes and products. Wiley-VHC, Weinheim
Matsushita K, Toyama H, Adachi O (1994) Respiratory chains and bioenergetics of acetic acid bacteria. Adv Microb Physiol 36:247–301. doi:10.1016/S0065-2911(08)60181-2
Mille Y, Beney L, Gervais P (2005) Compared tolerance to osmotic stress in various microorganisms: towards a survival prediction test. Biotechnol Bioeng 92(4):479–484. doi:10.1002/bit.20631
Mori H, Kobayashi T, Shimizu S (1981) High density production of sorbose from sorbitol by fed-batch culture with DO-stat. J Chem Eng Jpn 14(1):65–70. doi:10.1252/jcej.14.65
Nobel PS (1983) Introduction to biophysical plant physiology, 2 edn. San Fancisco
Olijve W, Kok JJ (1979) Analysis of growth of Gluconobacter oxydans in glucose containing media. Arch Microbiol 121(3):283–290. doi:10.1007/BF00425069
Olijve W, Kok JJ (1979) Analysis of the growth of Gluconobacter oxydans in chemostat cultures. Arch Microbiol 121(3):291–297. doi:10.1007/BF00425070
Peña C, Galindo E, Büchs J (2011) The viscosifying power, degree of acetylation and molecular mass of the alginate produced by Azotobacter vinelandii in shake flasks are determined by the oxygen transfer rate. Process Biochem 46(1):290–297. doi:10.1016/j.procbio.2010.08.025
Po HN, Senozan NM (2001) The Henderson-Hasselbalch equation: its history and limitations. J Chem Educ 78(11):1499. doi:10.1021/ed078p1499
Prior BA (1978) Effect of water activity on growth and respiration of Pseudomonas fluorescens. J Appl Bacteriol 44(1):97–106. doi:10.1111/j.1365-2672.1978.tb00780.x
Raspor P, Goranovic D (2008) Biotechnological applications of acetic acid bacteria. Crit Rev Biotechnol 28(2):101–124. doi:10.1080/07388550802046749
Record MT Jr, Courtenay ES, Cayley DS, Guttman HJ (1998) Responses of E. coli to osmotic stress: large changes in amounts of cytoplasmic solutes and water. Trends Biochem Sci 23(4):143–148. doi:10.1016/S0968-0004(98)01196-7
Richhardt J, Luchterhand B, Bringer S, Büchs J, Bott M (2013) Evidence for a key role of cytochrome bo3 oxidase in respiratory energy metabolism of Gluconobacter oxydans. J Bacteriol 195(18):4210–4220. doi:10.1128/JB.00470-13
Scheidle M, Dittrich B, Klinger J, Ikeda H, Klee D, Büchs J (2011) Controlling pH in shake flasks using polymer-based controlled-release discs with pre-determined release kinetics. BMC Biotechnol 11:25. doi:10.1186/1472-6750-11-25
Seletzky JM, Noack U, Hahn S, Knoll A, Amoabediny G, Büchs J (2007) An experimental comparison of respiration measuring techniques in fermenters and shake flasks: exhaust gas analyzer vs. RAMOS device vs. respirometer. J Ind Microbiol Biotechnol 34(2):123–130. doi:10.1007/s10295-006-0176-2
Shinagawa E, Ameyama M (1982) 2-Keto-d-gluconate dehydrogenase from Gluconobacter melanogenus, membrane-bound. Methods Enzymol
Shinagawa E, Matsushita K, Adachi O, Ameyama M (1981) Purification and characterization of 2-keto-d-gluconate dehydrogenase from Gluconobacter melanogenus. Agric Biol Chem 45(5):1079–1085. doi:10.1271/bbb1961.45.1079
Shinagawa E, Matsushita K, Adachi O, Ameyama M (1984) D-gluconate dehydrogenase, 2-keto-d-gluconate yielding, from Gluconobacter dioxyacetonicus—purification and characterization. Agric Biol Chem 48(6):1517–1522
Sievers M, Swings J, Garrity G, Brenner D, Krieg N, Staley J (2005) The genus gluconobacter. Springer, New York
Silberbach M, Maier B, Zimmermann M, Büchs J (2003) Glucose oxidation by Gluconobacter oxydans: characterization in shaking-flasks, scale-up and optimization of the pH profile. Appl Microbiol Biotechnol 62(1):92–98. doi:10.1007/s00253-003-1222-x
Sonoyama T, Tani H, Matsuda K, Kageyama B, Tanimoto M, Kobayashi K, Yagi S, Kyotani H, Mitsushima K (1982) Production of 2-keto-l-gulonic acid from d-glucose by two-stage fermentation. Appl Environ Microbiol 43(5):1064–1069
Sterne R, Zentmyer G, Bingham F (1976) The effect of osmotic potential and specific ions on growth of Phytophthora cinnamomi. GROWTH 1(1):5–10
Weenk G, Olijve W, Harder W (1984) Ketogluconate formation by Gluconobacter species. Appl Biochem Biotechnol 20(6):400–405. doi:10.1007/BF00261942
Weuster-Botz D (2005) Parallel reactor systems for bioprocess development. Adv Biochem Eng Biotechnol 92:125–143. doi:10.1007/b98916
Weuster-Botz D, Altenbach-Rehm J, Arnold M (2001) Parallel substrate feeding and pH-control in shaking-flasks. Biochem Eng J 7(2):163–170. doi:10.1016/S1369-703X(00)00117-0
Woods D, Duniway J (1986) Some effects of water potential on growth, turgor, and respiration of Phytophthora cryptogea and Fusarium moniliforme. Phytopathology 76(11):1248–1254
Zavrel M, Bross D, Funke M, Büchs J, Spiess AC (2009) High-throughput screening for ionic liquids dissolving (ligno-)cellulose. Bioresour Technol 100(9):2580–2587. doi:10.1016/j.biortech.2008.11.052
Zou X, Guo X, Sun M (2009) pH control strategy in a shaken mini bioreactor for polysaccharide production by medicinal mushroom Phellinus linteus and its anti-hyperlipemia activity. Bioprocess Biosyst Eng 32(2):277–281. doi:10.1007/s00449-008-0241-5
Acknowledgments
This work was performed within the research network “Genomik Transfer”, funded by the Federal Ministry of Research and Education, Germany (FKZ 0315632B). The authors wish to thank Stephanie Bringer and Janine Richhardt from the Institute of Bio- and Geosciences at the Research Center Jülich (Germany) as well was Wolfgang Liebl, Armin Ehrenreich and David Kostner from the Department of Microbiology at the Technological University Munich (Germany) for providing the G. oxydans strains ATCC 621H and DSM 3504.
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Luchterhand, B., Fischöder, T., Grimm, A.R. et al. Quantifying the sensitivity of G. oxydans ATCC 621H and DSM 3504 to osmotic stress triggered by soluble buffers. J Ind Microbiol Biotechnol 42, 585–600 (2015). https://doi.org/10.1007/s10295-015-1588-7
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DOI: https://doi.org/10.1007/s10295-015-1588-7