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Functional Characterization of a Novel Marine Microbial GDSL Lipase and Its Utilization in the Resolution of (±)-1-Phenylethanol

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Abstract

A novel GDSL lipase (MT6) was cloned from the genome of Marinactinospora thermotolerans SCSIO 00652 identified from the South China Sea. MT6 showed its maximum identity of 59 % with a putative lipase from Nocardiopsis dassonville. MT6 was heterologously expressed in E. coli BL21(DE3) and further functionally characterized. MT6 could efficiently resolve racemic 1-phenylethanol and generate (R)-1-phenylethanol with high enantiomeric excess (99 %) and conversion rate (54 %) through transesterification reactions after process optimization. Our report was the first one report about the utilization of one GDSL lipase in the preparation of chiral chemicals by transesterification reactions, and the optical selectivity of MT6 was interestingly opposite to those of other common lipases. GDSL lipases represented by MT6 possess great potential for the generation of valuable chiral chemicals in industry.

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References

  1. Suan, C. L., & Sarmidi, M. R. (2004). Immobilised lipase-catalysed resolution of (R, S)-1-phenylethanol in recirculated packed bed reactor. Journal of Molecular Catalysis B: Enzymatic, 28, 111–119.

    Article  Google Scholar 

  2. Schofer, S. H., Kaftzik, N., Wasserscheid, P., & Kragl, U. (2001). Enzyme catalysis in ionic liquids: lipase catalysed kinetic resolution of 1-phenylethanol with improved enantioselectivity. Chemical Communications, 5, 425–426.

    Article  Google Scholar 

  3. Frings, K., Koch, M., & Hartmeier, W. (1999). Kinetic resolution of 1-phenyl ethanol with high enantioselectivity with native and immobilized lipase in organic solvents. Enzyme and Microbial Technology, 25, 303–309.

    Article  CAS  Google Scholar 

  4. Dudzik, A., Snoch, W., Borowiecki, P., Opalinska-Piskorz, J., Witko, M., Heider, J., & Szaleniec, M. (2015). Asymmetric reduction of ketones and beta-keto esters by (S)-1-phenylethanol dehydrogenase from denitrifying bacterium Aromatoleum aromaticum. Applied Microbiology and Biotechnology, 99, 5055–5069.

    Article  CAS  Google Scholar 

  5. Hoffken, H. W., Duong, M., Friedrich, T., Breuer, M., Hauer, B., Reinhardt, R., Rabus, R., & Heider, J. (2006). Crystal structure and enzyme kinetics of the (S)-specific 1-phenylethanol dehydrogenase of the denitrifying bacterium strain EbN1. Biochemistry, 45, 82–93.

    Article  Google Scholar 

  6. Kniemeyer, O., & Heider, J. (2001). (S)-1-Phenylethanol dehydrogenase of Azoarcus sp strain EbN1, an enzyme of anaerobic ethylbenzene catabolism. Archives of Microbiology, 176, 129–135.

    Article  CAS  Google Scholar 

  7. Lozano, P., De Diego, T., Larnicol, M., Vaultier, M., & Iborra, J. L. (2006). Chemoenzymatic dynamic kinetic resolution of rac-1-phenylethanol in ionic liquids and ionic liquids/supercritical carbon dioxide systems. Biotechnology Letters, 28, 1559–1565.

    Article  CAS  Google Scholar 

  8. Li, X., Xu, L., Wang, G. L., Zhang, H. J., & Yan, Y. J. (2013). Conformation studies on Burkholderia cenocepacia lipase via resolution of racemic 1-phenylethanol in non-aqueous medium and its process optimization. Process Biochemistry, 48, 1905–1913.

    Article  CAS  Google Scholar 

  9. Cao, Y., Zhuang, Y., Yao, C. J., Wu, B., & He, B. F. (2012). Purification and characterization of an organic solvent-stable lipase from Pseudomonas stutzeri LC2-8 and its application for efficient resolution of (R, S)-1-phenylethanol. Biochemical Engineering Journal, 64, 55–60.

    Article  CAS  Google Scholar 

  10. Akoh, C. C., Lee, G. C., Liaw, Y. C., Huang, T. H., & Shaw, J. F. (2004). GDSL family of serine esterases/lipases. Progress in Lipid Research, 43, 534–552.

    Article  CAS  Google Scholar 

  11. Talker-Huiber, D., Jose, J., Glieder, A., Pressnig, M., Stubenrauch, G., & Schwab, H. (2003). Esterase EstE from Xanthomonas vesicatoria (Xv_EstE) is an outer membrane protein capable of hydrolyzing long-chain polar esters. Applied Microbiology and Biotechnology, 61, 479–487.

    Article  CAS  Google Scholar 

  12. Rathelot, J., Julien, R., Canioni, P., Coeroli, C., & Sarda, L. (1975). Studies on Effect of Bile-Salt and Colipase on Enzymatic Lipolysis - Improved Method for Determination of Pancreatic Lipase and Colipase. Biochimie, 57, 1117–1122.

    Article  CAS  Google Scholar 

  13. Chen, C. S., Fujimoto, Y., Girdaukas, G., & Sih, C. J. (1982). Quantitative-Analyses of Biochemical Kinetic Resolutions of Enantiomers. Journal of the American Chemical Society, 104, 7294–7299.

    Article  CAS  Google Scholar 

  14. Tian, X. P., Tang, S. K., Dong, J. D., Zhang, Y. Q., Xu, L. H., Zhang, S., & Li, W. J. (2009). Marinactinospora thermotolerans gen. nov., sp nov., a marine actinomycete isolated from a sediment in the northern South China Sea. International Journal of Systematic and Evolutionary Microbiology, 59, 948–952.

    Article  CAS  Google Scholar 

  15. Kumari, A., & Gupta, R. (2015). Functional characterization of a novel aspartic acid rich lipase, TALipC, from Trichosporon asahii MSR54: solvent-dependent enantio inversion during esterification of 1-phenylethanol. Biotechnology Letters, 37, 121–130.

    Article  CAS  Google Scholar 

  16. Cui, C. X., Xie, R., Tao, Y. F., Zeng, Q. Q., & Chen, B. Q. (2015). Improving performance of Yarrowia lipolytica lipase lip2-catalyzed kinetic resolution of (R, S)-1-phenylethanol by solvent engineering. Biocatalysis and Biotransformation, 33, 38–43.

    Article  CAS  Google Scholar 

  17. Li, X., Huang, S. S., Xu, L., & Yan, Y. J. (2013). Improving activity and enantioselectivity of lipase via immobilization on macroporous resin for resolution of racemic 1-phenylethanol in non-aqueous medium. BMC Biotechnology, 13, 986–991.

    Google Scholar 

  18. Xue, P., Yan, X. H., & Wang, Z. (2007). Lipase immobilized on HOOC-MCF: A highly enantioselective catalyst for transesterification resolution of (R, S)-1-phenylethanol. Chinese Chemical Letters, 18, 929–932.

    Article  CAS  Google Scholar 

  19. de los Rios, A. P., van Rantwijk, F., & Sheldon, R. A. (2012). Effective resolution of 1-phenyl ethanol by Candida antarctica lipase B catalysed acylation with vinyl acetate in protic ionic liquids (PILs). Green Chemistry, 14, 1584–1588.

    Article  Google Scholar 

  20. Petrovskaya, L. E., Novototskaya-Vlasova, K. A., Kryukova, E. A., Rivkina, E. M., Dolgikh, D. A., & Kirpichnikov, M. P. (2015). Cell surface display of cold-active esterase EstPc with the use of a new autotransporter from Psychrobacter cryohalolentis K5(T). Extremophiles, 19, 161–170.

    Article  CAS  Google Scholar 

  21. Prive, F., Kaderbhai, N. N., Girdwood, S., Worgan, H. J., Pinloche, E., Scollan, N. D., Huws, S. A., & Newbold, C. J. (2013). Identification and Characterization of Three Novel Lipases Belonging to Families II and V from Anaerovibrio lipolyticus 5ST. PloS One, 8, 1–9.

    Article  Google Scholar 

  22. Bae, S. Y., Ryu, B. H., Jang, E., Kim, S., & Kim, T. D. (2013). Characterization and immobilization of a novel SGNH hydrolase (Est24) from Sinorhizobium meliloti. Applied Microbiology and Biotechnology, 97, 1637–1647.

    Article  CAS  Google Scholar 

  23. Okamura, Y., Kimura, T., Yokouchi, H., Meneses-Osorio, M., Katoh, M., Matsunaga, T., & Takeyama, H. (2010). Isolation and Characterization of a GDSL Esterase from the Metagenome of a Marine Sponge-associated Bacteria. Marine Biotechnology, 12, 395–402.

    Article  CAS  Google Scholar 

  24. Ding, J. M., Yu, T. T., Liang, L. M., Xie, Z. R., Yang, Y. J., Zhou, J. P., Xu, B., Li, J. J., & Huang, Z. X. (2014). Biochemical Characterization of a GDSL-Motif Esterase from Bacillus sp K91 with a New Putative Catalytic Mechanism. Journal of Microbiology and Biotechnology, 24, 1551–1558.

    Article  CAS  Google Scholar 

  25. Yang, Z. X., Zhang, Y., Shen, T. T., Xie, Y., Mao, Y. M., & Ji, C. N. (2013). Cloning, expression and biochemical characterization of a novel, moderately thermostable GDSL family esterase from Geobacillus thermodenitrificans T2. Journal of Bioscience and Bioengineering, 115, 133–137.

    Article  CAS  Google Scholar 

  26. Feller, G., Zekhnini, Z., LamotteBrasseur, J., & Gerday, C. (1997). Enzymes from cold-adapted microorganisms - The class C beta-lactamase from the Antarctic psychrophile Psychrobacter immobilis A5. European Journal of Biochemistry, 244, 186–191.

    Article  CAS  Google Scholar 

  27. Russell, R. J. M., Gerike, U., Danson, M. J., Hough, D. W., & Taylor, G. L. (1998). Structural adaptations of the cold-active citrate synthase from an Antarctic bacterium. Structure, 6, 351–361.

    Article  CAS  Google Scholar 

  28. Belcastro, M., Marino, T., Russo, N., & Toscano, M. (2005). Interaction of cysteine with Cu2+ and Group IIb (Zn2+, Cd2+, Hg2+) metal cations: a theoretical study. Journal of Mass Spectrometry, 40, 300–306.

    Article  CAS  Google Scholar 

  29. Cote, A., & Shareck, F. (2008). Cloning, purification and characterization of two lipases from Streptomyces coelicolor A3(2). Enzyme and Microbial Technology, 42(5), 381–388.

    Article  CAS  Google Scholar 

  30. Mander, P., Cho, S. S., Simkhada, J. R., Choi, Y. H., Park, D. J., & Yoo, J. C. (2012). An organic solvent-tolerant lipase from Streptomyces sp CS133 for enzymatic transesterification of vegetable oils in organic media. Process Biochemistry, 47, 635–642.

    Article  CAS  Google Scholar 

  31. Farrokh, P., Yakhchali, B., & Karkhane, A. A. (2014). Cloning and characterization of newly isolated lipase from Enterobacter sp. Bn12. Brazilian Journal of Microbiology, 45, 677–687.

    Article  CAS  Google Scholar 

Download references

Acknowledgments

We are grateful for the financial supports from the National Natural Science Foundation of China (No. 21302199), Strategic Priority Research Program of the Chinese Academy of Sciences (XDA11030404), Project “Engineering High-Performance Microorganisms for Advanced Bio-Based Manufacturing” from the Chinese Academy of Sciences (KGZD-EW-606) and Guangzhou Science and Technology Plan Projects (201510010012). We also would like to thank for the constant helping from Professor Jianhua Ju and Professor Changsheng Zhang.

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Authors and Affiliations

  1. Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People’s Republic of China

    Dun Deng, Yun Zhang, Aijun Sun, Jiayuan Liang & Yunfeng Hu

  2. University of Chinese Academy of Sciences, Beijing, 100049, People’s Republic of China

    Dun Deng

  3. Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People’s Republic of China

    Dun Deng, Yun Zhang, Aijun Sun, Jiayuan Liang & Yunfeng Hu

  4. South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Guangzhou, People’s Republic of China

    Yunfeng Hu

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  1. Dun Deng
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  2. Yun Zhang
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Correspondence to Yunfeng Hu.

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Deng, D., Zhang, Y., Sun, A. et al. Functional Characterization of a Novel Marine Microbial GDSL Lipase and Its Utilization in the Resolution of (±)-1-Phenylethanol. Appl Biochem Biotechnol 179, 75–93 (2016). https://doi.org/10.1007/s12010-016-1980-4

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  • DOI: https://doi.org/10.1007/s12010-016-1980-4

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