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Comparison of Xylanases of Various Origin Obtained in the Expression System of Pichia pastoris: Gene Expression, Biochemical Characteristics, and Biotechnological Potential

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Abstract

Endo-1,4-β-xylanases from Bacillus pumilus VKPM В-7975, Paenibacillus polymyxa VKPM В-3015, Thermomyces lanuginosus VKPM F-224, and Schizophyllum commune VKPM F-433, which are members of a family of 11 glycohydrolases, have been obtained in the expression system of Pichia pastoris. Technologically valuable characteristics of the recombinant hydrolases, such as the specific activity, pH and thermal stability, temperature and pH optima, resistance to digestive enzymes and xylanase protein inhibitors from cereals, and substrate specificity were studied. It was shown that the enzyme from Paenibacillus polymyxa has the highest biotechnological potential for the creation of xylanases based on the recombinant Pichia pastoris yeast strain for their use in fodder production.

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REFERENCES

  1. Ebringerova, A. and Heinze, T., Xylan and xylan derivatives – biopolymers with valuable properties. 1: Naturally occurring xylans structures, isolation procedures and properties, Macromol. Rapid. Commun., 2000, vol. 21, no. 9, pp. 542–556. https://doi.org/10.1002/1521-3927(20000601)21:9<542::AID-MARC542>3.0.CO;2-7

    Article  CAS  Google Scholar 

  2. Coughlan, M.P. and Hazlewood, G.P., β-1,4-D-Xylan-degrading enzyme systems, biochemistry, molecular biology and applications, Biotechnol. Appl. Biochem., 1993, vol. 5, no. 17, pp. 259–289.

    Google Scholar 

  3. Kulkarni, N., Shendye, A., and Rao, M., Molecular and biotechnological aspects of xylanases, FEMS. Microbiol. Rev., 1999, vol. 5, no. 23, pp. 411–456.

    Article  Google Scholar 

  4. Coughlan, M.P. and Hazlewood, G.P., β-1,4-D-Xylan-degrading enzyme systems, biochemistry, molecular biology and applications, Biotechnol. Appl. Biochem., 1993, vol. 5, no. 17, pp. 259–289.

    Google Scholar 

  5. Subramaniyan, S. and Prema, P., Biotechnology of microbial xylanases: enzymology, molecular biology, and application, Crit. Rev. Biotechnol., 2002, vol. 22, pp. 33–64. https://doi.org/10.1080/07388550290789450

    Article  CAS  PubMed  Google Scholar 

  6. Harris, A.D. and Ramalingam, C., Xylanases and its application in food industry: a review, J. Exp. Sci., 2010, vol. 1, no. 7, pp. 1–11.

    Google Scholar 

  7. Beg, Q.K., Kapoor, M., Mahajan, L., and Hoondal, G.S., Microbial xylanases and their industrial applications: a review, Appl. Microbiol. Biotechnol., 2001, vol. 56, pp. 326–338.

    Article  CAS  Google Scholar 

  8. Gellissen, G., Heterologous protein production in methylotrophic yeasts, Appl. Microbiol. Biotechnol., 2000, vol. 54, no. 6, pp. 741–750.

    Article  CAS  Google Scholar 

  9. Cereghino, J.L. and Cregg, J.M., Heterologous protein expression in the methylotrophic yeast Pichia pastoris,FEMS. Microbiol. Rev., 2000, vol. 24, pp. 45–66. https://doi.org/10.1111/j.1574-6976.2000.tb00532.x

    Article  CAS  PubMed  Google Scholar 

  10. Sumner J.B., Somers G.F. Dinitrosalicylic method for glucose, in Laboratory Expression in Biological Chemistry, New York: Academic, 1949.

    Google Scholar 

  11. Teixeira, M.C., Alcazar, A.C., and Princi, E., Characterization of alkaline xylanases from Bacillus pumilus,Braz. J. Microbiol., 2000, vol. 31, pp. 90–94. https://doi.org/10.1590/S1517-83822000000200005

    Article  Google Scholar 

  12. Maha, T.H. Emam, KarimaA., et al., Cloning, sequencing and expression of the xylanase gene from Bacillus pumilus GH in Escherichia coli,Int. J. Chem. Tech. Res., 2016, vol. 9, no. 12, pp. 114–124.

    Google Scholar 

  13. Yang, R.C.A., Mackenzie, C.R., Bilous, D., et al., Molecular cloning and expression of a xylanase gene from Bacillus polymyxa in Escherichia coli,Appl. Environ. M-icrobiol., 1988, vol. 54, no. 4, pp. 1023–1029.

    CAS  Google Scholar 

  14. Morales, P., Madarro, A., Perez-Gonzalez, J.A., et al., Purification and characterization of alkaline xylanases from Bacillus polymyxa,Appl. Environ. Microbiol., 1993, vol. 59, no. 5, pp. 1376–1382.

    CAS  PubMed  PubMed Central  Google Scholar 

  15. Khucharoenphaisan, K. and Sinma, K., β-Xylanase from Thermomyces lanuginosus and its biobleaching application, Pakistan J. Biol. Sci., 2010, vol. 13, no. 11, pp. 513–526.

    Article  CAS  Google Scholar 

  16. Oku, T., Roy, C., Watsona, D.C., et al., Amino acid sequence and thermostability of xylanase A from Schizophyllum commune, FEBS Lett., 1993, vol. 334, no. 3, pp. 296–300.

    Article  CAS  Google Scholar 

  17. Paice, G., Jurasek, L., Carpenter, M.R., and Smillie, L.B., Production, characterization, and partial amino acid sequence of xylanase A from Schizophyllum commune,Appl. Environ. Microbiol., 1978, vol. 36, no. 6, pp. 802–808.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Fernandes, P., Enzymes in food processing: a condensed overview on strategies for better biocatalysts, Enzyme Res., 2010, vol. 2010, pp. 3–21. https://doi.org/10.4061/2010/862537

    Article  CAS  Google Scholar 

  19. Bustany, A.Z., The effect of pelleting an enzyme-supplemented barley-based broiler diet, Anim. Feed Sci. Technol., 1996, vol. 58, no. 3, pp. 283–288.

    Article  Google Scholar 

  20. Gusakov, A.V., Proteinaceous inhibitors of microbial xylanases, Biochemistry (Moscow), 2010, vol. 75, no. 10, pp. 1185–1199.

    CAS  PubMed  Google Scholar 

  21. Naatsaari, L., Mistlberger, B., Ruth, C., et al., Deletion of the Pichia pastoris KU70 homologue facilitates platform strain generation for gene expression and synthetic biology, PLoS One, 2012, vol. 7. e. 39720. https://doi.org/10.1371/journal.pone.0039720

    Article  Google Scholar 

  22. Sagt, B., Kleizen, R., Verwaal, M.D.M., et al., Introduction of an N-Glycosylation site increases secretion of heterologous proteins in yeasts, Appl. Environ. Mi-crobiol., 2000, vol. 66, no. 11, pp. 4940–4944.

    Article  CAS  Google Scholar 

  23. Minghai, H. and Xiaobin, Y., Enhanced expression of heterologous proteins in yeast cells via the modification of N-glycosylation sites. Bioengineered, 2015, vol. 6, no. 2, pp. 115–118. https://doi.org/10.1080/21655979.2015.1011031

    Article  CAS  Google Scholar 

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Funding

The work was supported by the Ministry of Education and Science of the Russian Federation (Unique Project Identifier RFMEFI60717X0180) with the Multipurpose Scientific Installation of the All-Russian Collection of Industrial Microorganisms of the National Bioresource Center, Kurchatov Institute (GOSNIIGENETIKA).

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Authors and Affiliations

  1. State Research Institute for Genetics and Selection of Industrial Microorganisms of the National Research Center, Kurchatov Institute (GOSNIIGENETIKA, NRC, Kurchatov Institute), 117545, Moscow, Russia

    A. N. Kalinina, L. N. Borshchevskaya, T. L. Gordeeva & S. P. Sineoky

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  1. A. N. Kalinina
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  2. L. N. Borshchevskaya
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  3. T. L. Gordeeva
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  4. S. P. Sineoky
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Correspondence to L. N. Borshchevskaya.

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The authors declare that they have no conflict of interest. This article does not contain any studies involving animals or human participants performed by any of the authors.

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Translated by I. Gordon

Abbreviations: CMC, carboxymethyl cellulose; GH11, family of 11 glycohydrolases; PAGE, polyacrylamide gel electrophoresis; SDS, sodium lauryl(dodecyl) sulphate; SDS-PAGE, protein PAGE in the occurrence of SDS; VKPM, All-Russia Collection of Industrial Microorganisms.

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Kalinina, A.N., Borshchevskaya, L.N., Gordeeva, T.L. et al. Comparison of Xylanases of Various Origin Obtained in the Expression System of Pichia pastoris: Gene Expression, Biochemical Characteristics, and Biotechnological Potential. Appl Biochem Microbiol 55, 733–740 (2019). https://doi.org/10.1134/S0003683819070044

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