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Efficient preparation of pseudoalteromone A from marine Pseudoalteromonas rubra QD1-2 by combination of response surface methodology and high-speed counter-current chromatography: a comparison with high-performance liquid chromatography

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Abstract

Pseudoalteromone A (PA) is a cytotoxic and anti-inflammatory ubiquinone discovered recently from a marine bacterium Pseudoalteromonas sp. CGH2XX. In order to meet its sample supply for further in vivo pharmacological investigation, an efficient method was developed for the preparation of PA by combination of response surface methodology (RSM) and high-speed counter-current chromatography (HSCCC) from marine bacterium P. rubra QD1-2. First, optimization of culture conditions was studied by the RSM to enhance PA production. The results indicated that the optimal cultivation condition was peptone (2.21 g/l), yeast extract (3.125 g/l), glucose (0.125 g/l), KBr (0.02 g/l), inoculum size (6.5 %), medium volume (595 ml), initial pH value (7.0), temperature (28 °C). Under the optimized fermentation condition, PA production was 1.04 mg/l with 14.8-fold increase comparing to 0.07 mg/l under original standard fermentation condition. The PA production was further investigated using a 14-l jar fermenter. Compared to the flask culture, P. rubra QD1-2 offered 45 % increase of PA production at 1.51 mg/l. Then, a rapid and efficient method for the separation and purification of PA from crude culture extract was developed using HSCCC. The two-phase solvent system used for HSCCC separation was composed of n-hexane–ethyl acetate–methanol–water (5:5:9:5, v/v/v/v). The isolation was accomplished within 100 min, and the purity of PA was over 95 %. The recovery of the process was 93 %.

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References

  • Beigi L, Karbalaei-Heidari HR, Kharrati-Kopaei M (2012) Optimization of an extracellular zinc-metalloprotease (SVP2) expression in Escherichia coli BL21 (DE3) using response surface methodology. Protein Expr Purif 84:161–166

    Article  CAS  PubMed  Google Scholar 

  • Bhatnagar I, Kim SK (2010) Immense essence of excellence: marine microbial bioactive compounds. Mar Drugs 8:2673–2701

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Bhavsar K, Gujar P, Shah P, Kumar VR, Khire JM (2013) Combinatorial approach of statistical optimization and mutagenesis for improved production of acidic phytase by Aspergillus niger NCIM 563 under submerged fermentation condition. Appl Microbiol Biotechnol 97:673–679

    Article  CAS  PubMed  Google Scholar 

  • Chen Y, Lu M, Chang Y, Hwang T, Wang W, Weng C, Kuo J, Sung P (2012) Pseudoalteromone A: a novel bioactive ubiquinone from a marine bacterium Pseudoalteromonas sp. CGH2XX (Pseudoalteromonadaceae). Tetrahedron Lett 53:1675–1677

    Article  CAS  Google Scholar 

  • Dai X, Huang Q, Zhou B, Gong Z, Liu Z, Shi S (2013) Preparative isolation and purification of seven main antioxidants from Eucommia ulmoides Oliv. (Du-zhong) leaves using HSCCC guided by DPPH-HPLC experiment. Food Chem 139:563–570

    Article  CAS  PubMed  Google Scholar 

  • Gao H, Liu M, Liu J, Dai H, Zhou X, Liu X, Zhuo Y, Zhang W, Zhang L (2009) Medium optimization for the production of avermectin B1a by Streptomyces avermitilis 14-12A using response surface methodology. Bioresour Technol 100:4012–4016

    Article  CAS  PubMed  Google Scholar 

  • Guo Z, Shen L, Ji Z, Wu W (2012) Enhanced production of a novel cyclic hexapeptide antibiotic (NW-G01) by Streptomyces alboflavus 313 using response surface methodology. Int J Mol Sci 13:5230–5241

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • He S, Wang H, Yan X, Zhu P, Chen J, Yang R (2013) Preparative isolation and purification of macrolactin antibiotics from marine bacterium Bacillus amyloliquefaciens using high-speed counter-current chromatography in stepwise elution mode. J Chromatogr A 1272:15–19

    Article  CAS  PubMed  Google Scholar 

  • Ito Y (2005) Golden rules and pitfalls in selecting optimum conditions for high-speed counter-current chromatography. J Chromatogr A 1065:145–168

    Article  CAS  PubMed  Google Scholar 

  • Liu R, Chu X, Sun A, Kong L (2005) Preparative isolation and purification of alkaloids from the Chinese medicinal herb Evodia rutaecarpa (Juss.) Benth by high-speed counter-current chromatography. J Chromatogr A 1074:139–144

    Article  CAS  PubMed  Google Scholar 

  • Maiti B, Rathore A, Srivastava S, Shekhawat M, Srivastava P (2011) Optimization of process parameters for ethanol production from sugar cane molasses by Zymomonas mobilis using response surface methodology and genetic algorithm. Appl Microbiol Biotechnol 90:385–395

    Article  CAS  PubMed  Google Scholar 

  • Mishra P, Kshirsagar PR, Nilegaonkar SS, Singh SK (2012) Statistical optimization of medium components for production of extracellular chitinase by Basidiobolus ranarum: a novel biocontrol agent against plant pathogenic fungi. J Basic Microbiol 52:539–548

    Article  CAS  PubMed  Google Scholar 

  • Montaser R, Luesch H (2011) Marine natural products: a new wave of drugs? Futur Med Chem 3:1475–1489

    Article  CAS  Google Scholar 

  • Newman DJ, Cragg GM (2012) Natural products as sources of new drugs over the 30 years from 1981 to 2010. J Nat Prod 75:311–335

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Papaneophytou CP, Kontopidis GA (2012) Optimization of TNF-alpha overexpression in Escherichia coli using response surface methodology: purification of the protein and oligomerization studies. Protein Expr Purif 86:35–44

    Article  CAS  PubMed  Google Scholar 

  • Peng J, Fan G, Qu L, Zhou X, Wu Y (2005) Application of preparative high-speed counter-current chromatography for isolation and separation of schizandrin and gomisin A from Schisandra chinensis. J Chromatogr A 1082:203–207

    Article  CAS  PubMed  Google Scholar 

  • Simmons TL, Coates RC, Clark BR, Engene N, Gonzalez D, Esquenazi E, Dorrestein PC, Gerwick WH (2008) Biosynthetic origin of natural products isolated from marine microorganism–invertebrate assemblages. Proc Natl Acad Sci U S A 105:4587–4594

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Su WT, Chen WJ, Lin YF (2009) Optimizing emulsan production of A. venetianus RAG-1 using response surface methodology. Appl Microbiol Biotechnol 84:271–279

    Article  CAS  PubMed  Google Scholar 

  • Sutherland I, Fisher D (2009) Role of counter-current chromatography in the modernisation of Chinese herbal medicines. J Chromatogr A 1216:740–753

    Article  CAS  PubMed  Google Scholar 

  • Tian J, Ye X, Shang Y, Deng Y, He K, Li X (2012) Preparative isolation and purification of harpagoside and angroside C from the root of Scrophularia ningpoensis Hemsley by high-speed counter-current chromatography. J Sep Sci 35:2659–2664

    Article  CAS  PubMed  Google Scholar 

  • Tuncer M, Kuru A, Isikli M, Sahin N, Celenk FG (2004) Optimization of extracellular endoxylanase, endoglucanase and peroxidase production by Streptomyces sp. F2621 isolated in Turkey. J Appl Microbiol 97:783–791

    Article  CAS  PubMed  Google Scholar 

  • Wang Q, Hou Y, Xu Z, Miao J, Li G (2008) Optimization of cold-active protease production by the psychrophilic bacterium Colwellia sp. NJ341 with response surface methodology. Bioresour Technol 99:1926–1931

    Article  CAS  PubMed  Google Scholar 

  • Wang J, Wen Y, Chen X, Lin Y, Zhou J, Xie Y, Wang H, Jiang H, Zheng W (2010) Preparative separation of six antimycin A components from antimycin fermentation broth by high-speed counter-current chromatography. J Chromatogr A 1217:5687–5692

    Article  CAS  PubMed  Google Scholar 

  • Wang X, Dong H, Liu Y, Yang B, Wang X, Huang L (2011) Application of high-speed counter-current chromatography for preparative separation of cyclic peptides from Vaccaria segetalis. J Chromatogr B Anal Technol Biomed Life Sci 879:811–814

    CAS  Google Scholar 

  • Wang Q, Song F, Xiao X, Huang P, Li L, Monte A, Abdel-Mageed WM, Wang J, Guo H, He W, Xie F, Dai H, Liu M, Chen C, Xu H, Liu M, Piggott AM, Liu X, Capon RJ, Zhang L (2013) Abyssomicins from the south China sea deep-sea sediment Verrucosispora sp.: natural thioether michael addition adducts as antitubercular prodrugs. Angew Chem Int Ed 52:1231–1234

    Article  CAS  Google Scholar 

  • Xiao X, Si X, Yuan Z, Xu X, Li G (2012) Isolation of fucoxanthin from edible brown algae by microwave-assisted extraction coupled with high-speed countercurrent chromatography. J Sep Sci 35:2313–2317

    Article  CAS  PubMed  Google Scholar 

  • Xiong ZQ, Tu XR, Tu GQ (2008) Optimization of medium composition for actinomycin X2 production by Streptomyces spp JAU4234 using response surface methodology. J Ind Microbiol Biotechnol 35:729–734

    Article  CAS  PubMed  Google Scholar 

  • Xiong ZQ, Wang JF, Hao YY, Wang Y (2013) Recent advances in the discovery and development of marine microbial natural products. Mar Drugs 11:700–717

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  • Zhao J, Mou Y, Shan T, Li Y, Lu S, Zhou L (2012) Preparative separation of helvolic acid from the endophytic fungus Pichia guilliermondii Ppf9 by high-speed counter-current chromatography. World J Microbiol Biotechnol 28:835–840

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This work was financially supported by National Natural Science Foundation of China (41306134), 863 Program of China (2013AA092902), Zhejiang Provincial Natural Science Foundation of China (LQ13B020004), Ph.D. Programs Foundation of Ministry of Education of China (20133305120007), Natural Science Foundation of Ningbo (2013A610278), Zhejiang Marine Biotechnology Innovation Team (2012R10029-2), Ningbo Marine Algae Biotechnology Team (2011B81007), Scientific Research Foundation for Returned Scholars of ZJHRSS and K.C. Wong Magna Fund in Ningbo University.

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Correspondence to Shan He or Xiaojun Yan.

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Ding, L., He, S. & Yan, X. Efficient preparation of pseudoalteromone A from marine Pseudoalteromonas rubra QD1-2 by combination of response surface methodology and high-speed counter-current chromatography: a comparison with high-performance liquid chromatography. Appl Microbiol Biotechnol 98, 4369–4377 (2014). https://doi.org/10.1007/s00253-014-5530-0

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