Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column

doi:10.1016/S1004-9541(13)60514-5

Chin.J.Chem.Eng. ›› 2013, Vol. 21 ›› Issue (6): 663-669.DOI: 10.1016/S1004-9541(13)60514-5

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Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column

SHEN Shaochuan^1,2, WANG Liangyan¹, SUN Zongtao³, LI Mingfeng¹, LIU Chengzhi¹, TIAN Bing¹, YUN Junxian², HUA Yuejin¹

1 Key Laboratory for Nuclear-Agricultural Sciences of Chinese Ministry of Agriculture and Zhejiang Province, Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China;
2 State Key Laboratory Breeding Base of Green Chemistry Synthesis Technology, College of Chemical Engineering and Materials Science, Zhejiang University of Technology, Hangzhou 310032, China;
3 Department of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2012-04-05 Revised:2012-12-30 Online:2013-07-03 Published:2013-06-28
Contact: HUA Yuejin
Supported by:
Supported by the National Natural Science Foundation of China (30830006, 20876145, 21036005), the International Science & Technology Cooperation Program from the Ministry of Science and Technology of China (1017), the Special Fund for Agroscientific Research in the Public Interest (201103007), the Fundamental Research Funds for the Central Universities and the Natural Science Foundation of Zhejiang Province (Y4080326, Y407366).

Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column

沈绍传^1,2, 王梁燕¹, 孙宗涛³, 李铭峰¹, 刘程智¹, 田兵¹, 贠军贤², 华跃进¹

1 Key Laboratory for Nuclear-Agricultural Sciences of Chinese Ministry of Agriculture and Zhejiang Province, Institute of Nuclear-Agricultural Sciences, Zhejiang University, Hangzhou 310029, China;
2 State Key Laboratory Breeding Base of Green Chemistry Synthesis Technology, College of Chemical Engineering and Materials Science, Zhejiang University of Technology, Hangzhou 310032, China;
3 Department of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

通讯作者: HUA Yuejin
基金资助:
Supported by the National Natural Science Foundation of China (30830006, 20876145, 21036005), the International Science & Technology Cooperation Program from the Ministry of Science and Technology of China (1017), the Special Fund for Agroscientific Research in the Public Interest (201103007), the Fundamental Research Funds for the Central Universities and the Natural Science Foundation of Zhejiang Province (Y4080326, Y407366).

Abstract

Abstract: Geranylgeranyl diphosphate synthase (GGPPS) plays a key role in the biosynthesis of antioxidative carotenoid from the extremely radioresistant bacterium Deinococcus radiodurans. In this work, the recombinant GGPPS expressed in Escherichia coli by cloning and transforming the gene dr1395 of D. radiodurans was isolated rapidly by an immobilized metal affinity supermacroporous cryogel, i.e., Cu²⁺-iminodiacetic acid (IDA)-cryogel. The properties of the Cu²⁺-IDA-cryogel were characterized using capillary-based mathematical model and experimental measurements. The obtained protein samples were analyzed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The porosity of the present Cu²⁺-IDA-cryogel is 90.4% and the water permeability is 5.04×10^-12 m². From the capillary-based model, this cryogel presents a slightly wide normal pore (capillary) size distribution with the mean diameter of 55.2 μm, the standard deviation of 28.0 μm and the half of skeleton wall thickness of 2.8 μm. The pore size distribute from about 10 to 141 μm and the effective tortuosity of these capillary pores increases from 2.60 to 9.05. The isolation of the GGPPS from cell homogenate can be achieved at the flow velocity of 3.40×10^-4 m·s^-1 by the Cu²⁺-IDA-cryogel bed. High-purity GGPPS (about 91.4%) is obtained according to the SDS-PAGE analysis of the elution samples, indicating that the present method is a promising, simple and effective approach to isolate GGPPS from cell homogenate of engineering strains.

Key words: chromatography, separation, protein, modeling, bacterium

摘要： Geranylgeranyl diphosphate synthase (GGPPS) plays a key role in the biosynthesis of antioxidative carotenoid from the extremely radioresistant bacterium Deinococcus radiodurans. In this work, the recombinant GGPPS expressed in Escherichia coli by cloning and transforming the gene dr1395 of D. radiodurans was isolated rapidly by an immobilized metal affinity supermacroporous cryogel, i.e., Cu²⁺-iminodiacetic acid (IDA)-cryogel. The properties of the Cu²⁺-IDA-cryogel were characterized using capillary-based mathematical model and experimental measurements. The obtained protein samples were analyzed by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The porosity of the present Cu²⁺-IDA-cryogel is 90.4% and the water permeability is 5.04×10^-12 m². From the capillary-based model, this cryogel presents a slightly wide normal pore (capillary) size distribution with the mean diameter of 55.2 μm, the standard deviation of 28.0 μm and the half of skeleton wall thickness of 2.8 μm. The pore size distribute from about 10 to 141 μm and the effective tortuosity of these capillary pores increases from 2.60 to 9.05. The isolation of the GGPPS from cell homogenate can be achieved at the flow velocity of 3.40×10^-4 m·s^-1 by the Cu²⁺-IDA-cryogel bed. High-purity GGPPS (about 91.4%) is obtained according to the SDS-PAGE analysis of the elution samples, indicating that the present method is a promising, simple and effective approach to isolate GGPPS from cell homogenate of engineering strains.

关键词: chromatography, separation, protein, modeling, bacterium

SHEN Shaochuan, WANG Liangyan, SUN Zongtao, LI Mingfeng, LIU Chengzhi, TIAN Bing, YUN Junxian, HUA Yuejin . Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column[J]. Chin.J.Chem.Eng., 2013, 21(6): 663-669.

沈绍传, 王梁燕, 孙宗涛, 李铭峰, 刘程智, 田兵, 贠军贤, 华跃进. Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column[J]. Chinese Journal of Chemical Engineering, 2013, 21(6): 663-669.

References

1 Minton, K.W., “DNA repair in the extremely radioresistant bacterium Deinococcus radiodurans”, Mol. Microbiol., 13, 9-15 (1994).

2 Cox, M.M., Battista, J.R., “Deinococcus radiodurans—the consummate survivor”, Nat. Rev. Microbiol., 3, 882-892 (2005).

3 Battista, J.R., Earl, A.M., Park, M.J., “Why is Deinococcus radiodurans so resistant to ionizing radiation?”, Trends Microbiol., 7, 362-365 (1999).

4 Lemee, L., Peuchant, E., Clerc, M., Brunner, M., Pfander, H., “Deinoxanthin: A new carotenoid isolated from Deinococcus radiodurans”, Tetrahedron, 53, 919-926 (1997).

5 Tian, B., Hua, Y.J., “Carotenoid biosynthesis in extremophilicDeinococcus-Thermus bacteria”, Trends Microbiol., 18, 512-520 (2010).

6 Slade, D., Radman, M., “Oxidative stress resistance in Deinococcus radiodurans”, Microbiol. Mol. Biol. R., 75, 133-191 (2011).

7 Tian, B., Xu, Z.J., Sun, Z.T., Lin, J., Hua, Y.J., “Evaluation of the antioxidant effects of carotenoids from Deinococcus radiodurans through targeted mutagenesis, chemiluminescence, and DNA damage analyses”, BBA—Gen. Subjects, 1770, 902-911 (2007).

8 Xu, Z.J., Tian, B., Sun, Z.T., Lin, J., Hua, Y.J., “Identification and functional analysis of a phytoene desaturase gene from the extremely radioresistant bacterium Deinococcus radiodurans”, Microbiol. —SGM, 153, 1642-1652 (2007).

9 Tian, B., Sun, Z.T., Xu, Z.J., Shen, S.C., Wang, H., Hua, Y.J., “Carotenoid 3′,4′-desaturase is involved in carotenoid biosynthesis in the radioresistant bacterium Deinococcus radiodurans”, Microbiol. —SGM, 154, 3697-3706 (2008).

10 Sun, Z.T., Shen, S.C., Wang, C., Wang, H., Hu, Y.P., Jiao, J.D., Ma, T.T., Tian, B., Hua, Y.J., “A novel carotenoid 1,2-hydratase (CruF) from two species of the non-photosynthetic bacterium Deinococcus”, Microbiol. —SGM, 155, 2775-2783 (2009).

11 Math, S.K., Hearst, J.E., Poulter, C.D., “The crtE gene in Erwinia herbicola encodes geranylgeranyl diphosphate synthase”, P. Natl. Acad. Sci. USA, 89, 6761-6764 (1992).

12 Miyagi, Y., Matsumura, Y., Sagami, H., “Human geranylgeranyl diphosphate synthase is an octamer in solution”, J. Biochem., 142, 377-381 (2007).

13 Makarova, K.S., Aravind, L., Wolf, Y.I., Tatusov, R.L., Minton, K.W., Koonin, E.V., Daly, M.J., “Genome of the extremely radiation-resistant bacterium Deinococcus radiodurans viewed from the perspective of comparative genomics”, Microbiol. Mol. Biol. R., 65, 44-79 (2001).

14 Ghosal, D., Omelchenko, M.V., Gaidamakova, E.K., Matrosova, V.Y., Vasilenko, A., Venkateswaran, A., Zhai, M., Kostandarithes, H., Brim, H., Makarova, K., “How radiation kills cells: survival of Deinococcus radiodurans and Shewanella oneidensis under oxidative stress”, FEMS Microbiol. Rev., 29, 361-375 (2005).

15 Lozinsky, V.I., Plieva, F.M., Galaev, I.Y., Mattiasson, B., “The potential of polymeric cryogels in bioseparation”, Bioseparation, 10, 163-188 (2002).

16 Lozinsky, V.I., Galaev, I.Y., Plieva, F.M., Savina, I.N., Jungvid, H., Mattiasson, B., “Polymeric cryogels as promising materials of biotechnological interest”, Trends Biotechnol., 21, 445-451 (2003).

17 Plieva, F.M., Galaev, I.Y., Mattiasson, B., “Macroporous gels prepared at subzero temperatures as novel materials for chromatography of particulate-containing fluids and cell culture applications”, J. Sep. Sci., 30, 1657-1671 (2007).

18 Lozinsky, V.I., “Polymeric cryogels as a new family of macroporous and super-macroporous materials for biotechnological purposes”, Russ. Chem. Bull., 57, 1015-1032 (2008).

19 Plieva, F.M., Kirsebom, H., Mattiasson, B., “Preparation of macroporous cryostructurated gel monoliths, their characterization and main applications”, J. Sep. Sci., 34, 2164-2172 (2011).

20 Kirsebom, H., Galaev, I.Y., Mattiasson, B., “Stimuli-responsive polymers in the 21st century: Elaborated architecture to achieve high sensitivity, fast response, and robust behavior”, J. Polym. Sci. Pol. Phys., 49, 173-178 (2011).

21 Arvidsson, P., Plieva, F.M., Savina, I.N., Lozinsky, V.I., Fexby, S., Bulow, L., Galaev, I.Y., Mattiasson, B., “Chromatography of microbial cells using continuous super-macroporous affinity and ion-exchange columns”, J. Chromatogr. A, 977, 27-38 (2002).

22 Plieva, F.M., Galaev, I.Y., Noppe, W., Mattiasson, B., “Cryogel applications in microbiology”, Trends Microbiol., 16, 543-551 (2008).

23 Yun, J.X., Shen, S.C., Chen, F., Yao, K.J., “One-step isolation of adenosine triphosphate from crude fermentation broth of Saccharomyces cerevisiae by anion-exchange chromatography using supermacroporous cryogel”, J. Chromatogr. B, 860, 57-62 (2007).

24 Wang, L.H., Shen, S.C., Yun, J.X., Yao, K.J., Yao, S.J., “Chromatographic separation of cytidine triphosphate from fermentation broth of yeast using anion-exchange cryogel”, J. Sep. Sci., 31, 689-695 (2008).

25 Chen, Y., Shen, S.C., Yun, J.X., Yao, K.J., “Isolation of ATP from a yeast fermentation broth using a cryogel column at high flow velocities”, J. Sep. Sci., 31, 3879-3883 (2008).

26 Sagami, H., Morita, Y., Ogura, K., “Purification and properties of geranylgeranyl-diphosphate synthase from bovine brain”, J. Biol. Chem., 269, 20561-20566 (1994).

27 Ogura, K., Shinka, T., Seto, S., “The purification and properties of geranylgeranyl pyrophosphate synthetase from pumpkin fruit”, J. Biochem., 72, 1101-1108 (1972).

28 Nishio, K., Nodake, Y., Hamada, K., Suto, K., Nakagawa, N., Kuramitsu, S., Miura, K., “Expression, purification, crystallization and preliminary X-ray studies of geranylgeranyl diphosphate synthase from Thermus thermophilus HB8”, Acta Crystallogr. D, 60, 178-180 (2004).

29 Laskaris, G., van der Heijden, R., Verpoorte, R., “Purification and partial characterisation of geranylgeranyl diphosphate synthase, from Taxus baccata cell cultures. An enzyme that regulates taxane biosynthesis”, Plant Sci., 153, 97-105 (2000).

30 Brinkhaus, F.L., Rilling, H.C., “Purification of geranylgeranyl diphosphate synthase from Phycomyces blakesleanus”, Arch. Biochem. Biophys., 266, 607-612 (1988).

31 Yao, K.J., Shen, S.C., Yun, J.X., Wang, L.H., He, X.J., Yu, X.M., “Preparation of polyacrylamide-based super-macroporous monolithic cryogel beds under freezing-temperature variation conditions”, Chem. Eng. Sci., 61, 6701-6708 (2006).

32 Wang, L.H., Shen, S.C., He, X.J., Yun, J.X., Yao, K.J., Yao, S.J., “Adsorption and elution behaviors of bovine serum albumin in metal-chelated affinity cryogel beds”, Biochem. Eng. J., 42, 237-242 (2008).

33 He, X.J., Yao, K.J., Shen, S.C., Yun, J.X., “Freezing characteristics of acrylamide-based aqueous solution used for the preparation of super-macroporous cryogels via cryo-copolymerization”, Chem. Eng. Sci., 62, 1334-1342 (2007).

34 Yao, K.J., Yun, J.X., Shen, S.C., Wang, L.H., He, X.J., Yu, X.M., “Characterization of a novel continuous super-macroporous monolithic cryogel embedded with nanoparticles for protein chromatography”, J. Chromatogr. A, 1109, 103-110 (2006).

35 Plieva, F.M., Savina, I.N., Deraz, S., Andersson, J., Galaev, I.Y., Mattiasson, B., “Characterization of super-macroporous monolithic polyacrylamide based matrices designed for chromatography of bioparticles”, J. Chromatogr. B, 807, 129-137 (2004).

36 Yun, J.X., Lin, D.Q., Yao, S.J., “Predictive modeling of protein adsorption along the bed height by taking into account the axial nonuniform liquid dispersion and particle classification in expanded beds”, J. Chromatogr. A, 1095, 16-26 (2005).

37 Yun, J.X., Jespersen, G.R., Kirsebom, H., Gustavsson, P.E., Mattiasson, B., Galaev, I.Y., “An improved capillary model for describing the microstructure characteristics, fluid hydrodynamics and breakthrough performance of proteins in cryogel beds”, J. Chromatogr. A, 1218, 5487-5497 (2011).

38 Yun, J.X., Kirsebom, H., Galaev, I.Y., Mattiasson, B., “Modeling of protein breakthrough performance in cryogel columns by taking into account the overall axial dispersion”, J. Sep. Sci., 32, 2601-2607 (2009).

39 Persson, P., Baybak, O., Plieva, F., Galaev, I.Y., Mattiasson, B., Nilsson, B., Axelsson, A., “Characterization of a continuous super- macroporous monolithic matrix for chromatographic separation of large bioparticles”, Biotechnol. Bioeng., 88, 224-236 (2004).

40 Yan, L.D., Shen, S.C., Yun, J.X., Yao, K.J., “Isolation of lysozyme from chicken egg white using polyacrylamide-based cation-exchange cryogel”, Chin. J. Chem. Eng., 19, 876-880 (2011).

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Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column

Separation of Recombinant Geranylgeranyl Diphosphate Synthase of Deinococcus radiodurans from Expressed Strain Cell Homogenate by Immobilized Metal Affinity Chromatography on a Characterized Monolithic Cryogel Column

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