Skip to main content
SpringerLink
Log in

Bamboo (Phyllostachys pubescens) as a Natural Support for Neutral Protease Immobilization

  • Published:
Applied Biochemistry and Biotechnology Aims and scope Submit manuscript

Abstract

Lignin polymers in bamboo (Phyllostachys pubescens) were decomposed into polyphenols at high temperatures and oxidized for the introduction of quinone groups from peroxidase extracted from bamboo shoots and catalysis of UV. According to the results of FT-IR spectra analysis, neutral proteases (NPs) can be immobilized on the oxidized lignin by covalent bonding formed by amine group and quinone group. The optimum condition for the immobilization of NPs on the bamboo bar was obtained at pH 7.0, 40 °C, and duration of 4 h; the amount of immobilized enzyme was up to 5 mg g−1 bamboo bar. The optimal pH for both free NP (FNP) and INP was approximately 7.0, and the maximum activity of INP was determined at 60 °C, whereas FNP presented maximum activity at 50 °C. The Km values of INP and FNP were determined as 0.773 and 0.843 mg ml−1, respectively; INP showed a lower Km value and Vmax, than FNP, which demonstrated that INP presented higher affinity to substrate. Compared to FNP, INP showed broader thermal and storage stability under the same trial condition. With respect to cost, INP presented considerable recycling efficiency for up to six consecutive cycles.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Wang, G., Xu, A., Wan, Y., & Li, Q. (2013). Purification and characterization of a new metallo-neutral protease for beer brewing from Bacillus amyloliquefaciens SYB-001. Applied Biochemistry and Biotechnology, 170(8), 2021–2033. https://doi.org/10.1007/s12010-013-0350-8

    Article  CAS  PubMed  Google Scholar 

  2. Wittrock, H., Jiang, M., Campbell, M., Jane, J., Anih, E., & Wang, E. (2008). A simplified isolation of high-amylose maize starch using neutral proteases. Starch-Stärke, 60(11), 601–608. https://doi.org/10.1002/star.200800223

    Article  CAS  Google Scholar 

  3. Szot, G., Lee, M., Tavakol, M., Lang, J., Dekovic, F., Kerlan, R., & Posselt, A. (2009). Successful clinical islet isolation using a GMP-manufactured collagenase and neutral protease. Transplantation, 88(6), 753–756. https://doi.org/10.1097/TP.0b013e3181b443ae

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Seabra, I., & Gil, M. (2007). Cotton gauze bandage: a support for protease immobilization for use in biomedical applications. Brazilian Journal of Pharmaceutical Sciences, 43, 535–542.

    CAS  Google Scholar 

  5. Gilani, S., Najafpour, G., Moghadamnia, A., & Kamaruddin, A. (2016). Stability of immobilized porcine pancreas lipase on mesoporous chitosan beads: a comparative study. Journal of Molecular Catalysis B: Enzymatic, 133, 144–153. https://doi.org/10.1016/j.molcatb.2016.08.005

    Article  CAS  Google Scholar 

  6. Fernandez-Lopez, L., Pedrero, S., Lopez-Carrobles, N., Gorines, B., Virgen-Ortíz, J., & Fernandez-Lafuente, R. (2017). Effect of protein load on stability of immobilized enzymes. Enzyme and Microbial Technology, 98, 18–25. https://doi.org/10.1016/j.enzmictec.2016.12.002

    Article  CAS  PubMed  Google Scholar 

  7. Al-Qodah, Z., Al-Shannag, M., Al-Busoul, M., Penchev, I., & Orfali, W. (2017). Immobilized enzymes bioreactors utilizing a magnetic field: a review. Biochemical Engineering Journal, 121, 94–106. https://doi.org/10.1016/j.bej.2017.02.003

    Article  CAS  Google Scholar 

  8. Jiang, Y., Sun, W., Wang, Y., Wang, W., Zhou, L., Gao, J., & Zhang, X. (2017). Protein-based inverse opals: a novel support for enzyme immobilization. Enzyme and Microbial Technology, 96, 42–46. https://doi.org/10.1016/j.enzmictec.2016.08.021

    Article  CAS  PubMed  Google Scholar 

  9. Liu, Y., & Chen, J. (2016). Enzyme immobilization on cellulose matrixes. Journal of Bioactive and Compatible Polymers, 31(6), 553–567. https://doi.org/10.1177/0883911516637377

    Article  CAS  Google Scholar 

  10. Catia, A., Laura, D., & Lidietta, G. (2017). Tyrosinase immobilized on a hydrophobic membrane. Biotechnology and Applied Biochemistry, 64, 92–99.

    Article  CAS  Google Scholar 

  11. Li, J., Du, Y., Sun, L., Liang, H., Feng, T., Wei, Y., & Yao, P. (2006). Chitosaneous hydrogel beads for immobilizing neutral protease for application in the preparation of low molecular weight chitosan and chito-oligomers. Journal of Applied Polymer Science, 101, 3742–3750.

    Google Scholar 

  12. Bavaro, T., Cattaneo, G., Serra, I., Benucci, I., Pregnolato, M., & Terreni, M. (2017). Immobilization of neutral protease from Bacillus subtilis for regioselective hydrolysis of acetylated nucleosides: application to capecitabine synthesis. Molecules, 21, 1–p16.

    Google Scholar 

  13. Li, W., Wen, H., Shi, Q. Q., & Zheng, G. G. (2016). Study on immobilization of (+)-lactamase using a new type of epoxy graphene oxide carrier. Process Biochemistry, 51(2), 270–276. https://doi.org/10.1016/j.procbio.2015.11.030

    Article  CAS  Google Scholar 

  14. Tang, Z., Qian, J., & Shi, L. (2007). Preparation of chitosan nanoparticles as carrier for immobilized enzyme. Applied Biochemistry and Biotechnology, 136(1), 77–96. https://doi.org/10.1007/BF02685940

    Article  CAS  PubMed  Google Scholar 

  15. Yang, D., Zhong, L., Yuan, T., Peng, X., & Sun, R. (2013). Studies on the structural characterization of lignin, hemicelluloses and cellulose fractionated by ionic liquid followed by alkaline extraction from bamboo. Industrial Crops and Products, 43, 141–149. https://doi.org/10.1016/j.indcrop.2012.07.024

    Article  CAS  Google Scholar 

  16. Gong, W. H., Ran, Z. X., Ye, F. Y., & Zhao, G. H. (2017). Lignin from bamboo shoot shells as an activator and novel immobilizing support for a-amylase. Food Chemistry, 228, 455–462. https://doi.org/10.1016/j.foodchem.2017.02.017

    Article  CAS  PubMed  Google Scholar 

  17. Park, S., Kim, S. H., Kim, J. H., Yu, H., Kim, H. J., Yang, Y. H., Kim, H., Kim, Y. H., Ha, S. H., & Lee, S. H. (2015). Application of cellulose/lignin hydrogel beads as novel supports for immobilizing lipase. Journal of Molecular Catalysis B: Enzyatic, 119, 33–39. https://doi.org/10.1016/j.molcatb.2015.05.014

    Article  CAS  Google Scholar 

  18. Zdarta, J., Klapiszewski, L., Wysokowski, M., Norman, M., Kolodziejczak-radzimska, A., Moszynski, D., Ehrlich, H., Maciejewski, H., Stelling, A. L., & Jesionowski, T. (2015). Chitin-lignin material as a novel matrix for enzyme immobilization. Marine Drugs, 13(4), 2424–2446. https://doi.org/10.3390/md13042424

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Ajitha, S., & Sugunan, S. (2010). Tuning mesoporous molecular sieve SBA-15 for the immobilization of α-amylase. Journal of Porous Materials, 17(3), 341–349. https://doi.org/10.1007/s10934-009-9298-z

    Article  CAS  Google Scholar 

  20. Fei, X. Y., Chen, S. Y., Liu, D., Huang, C. J., & Zhang, Y. C. (2016). Comparison of amino and epoxy functionalized SBA-15 used for carbonic anhydrase immobilization. Journal of Bioscience and Bioengineering, 122(3), 314–321. https://doi.org/10.1016/j.jbiosc.2016.02.004

    Article  CAS  PubMed  Google Scholar 

  21. Fan, L., Ruan, R., Liu, Y., Wang, Y., & Tu, C. (2015). Effects of extraction conditions on the characteristics of ethanol organosolv lignin from bamboo (Phyllostachys pubescens Mazel). BioResources, 10, 7998–8013.

    CAS  Google Scholar 

  22. Gao, G., Karaaslan, M., & Kadla, J. (2014). Hydrogen-bonding based reversible polymer networks based on Kraft lignin and poly(2-(Dimethylamino)ethyl methacrylate) series polymers. Macromolecular Materials and Engineering, 299(8), 990–1002. https://doi.org/10.1002/mame.201300364

    Article  CAS  Google Scholar 

  23. Satoshi, K., John, F., & Kadla. (2005). Kraft lignin/poly(ethylene oxide) blends: effect of lignin structure on miscibility and hydrogen bonding. Journal of Applied Polymer Science, 98, 1438–1444.

    Google Scholar 

  24. Chen, Z., & Wan, C. (2017). Biological valorization strategies for converting lignin into fuels and chemicals. Renewable and Sustainable Energy Reviews, 73, 610–621. https://doi.org/10.1016/j.rser.2017.01.166

    Article  CAS  Google Scholar 

  25. Dieste, A., Clavijo, L., Torres, A., Barbe, S., Oyarbide, I., Bruno, L., & Cassella, F. (2016). Lignin from Eucalyptus spp. kraft black liquor as biofuel. Energy & Fuels, 30(12), 10494–10498. https://doi.org/10.1021/acs.energyfuels.6b02086

    Article  CAS  Google Scholar 

  26. Salma, K. B. G., Lobna, M., Sana, K., Kalthoum, C., Imene, O., & Abdelwaheb, C. (2016). Antioxidant enzymes expression in Pseudomonas aeruginosa exposed to UV-C radiation. Journal of Basic Microbiology, 56(7), 736–740. https://doi.org/10.1002/jobm.201500753

    Article  CAS  PubMed  Google Scholar 

  27. Tang, Z., Qian, J., & Shi, L. (2006). Characterizations of immobilized neutral proteinase on chitosan nano-particles. Process Biochemistry, 41(5), 1193–1197. https://doi.org/10.1016/j.procbio.2005.11.015

    Article  CAS  Google Scholar 

  28. Gong, W. H., Xiang, Z. Y., Ye, F. Y., & Zhao, G. H. (2016). Composition and structure of an antioxidant acetic acid lignin isolated from shoot shell of bamboo (Dendrocalamus Latiforus). Industrial Crops and Products, 91, 340–349. https://doi.org/10.1016/j.indcrop.2016.07.023

    Article  CAS  Google Scholar 

  29. Ouyang, X., Ke, L., Qiu, X., Guo, Y., & Pang, Y. (2009). Sulfonation of alkali lignin and its potential use in dispersant for cement. Journal of Dispersion Science and Technology, 30(1), 1–6. https://doi.org/10.1080/01932690802473560

    Article  CAS  Google Scholar 

  30. Klinman, J., & Bonnot, F. (2004). Intrigues and intricacies of the biosynthetic pathways for the enzymatic quinocofactors: PQQ, TTQ, CTQ, TPQ, and LTQ. Chemical Reviews, 114, 4343–4365.

    Article  CAS  Google Scholar 

  31. Natasa, Z., Sekuljica • Nevena, Z., Prlainovic • Jelena, R., Jovanovic •Andrea, B., Stefanovic • Veljko, R., Djokic • Dusan, Z., Mijin • Zorica, D., & Knezevic, J. (2016). Immobilization of horseradish peroxidase onto kaolin. Bioprocess and Biosystems Engineering, 39, 461–472.

    Article  CAS  Google Scholar 

  32. Jiang, D., Long, S., Huang, J., Xiao, H., & Zhou, D. (2005). Immobilization of Pycnoporus sanguineus laccase on magnetic chitosan microspheres. Biochemical Engineering Journal, 25(1), 15–23. https://doi.org/10.1016/j.bej.2005.03.007

    Article  CAS  Google Scholar 

  33. Tyagi, C., Tomar, L. K., & Singh, H. (2009). Surface modification of cellulose filter paper by glycidyl methacrylate grafting for biomolecule immobilization: influence of grafting parameters and urease immobilization. Journal of Applied Polymer Science, 111(3), 1381–1390. https://doi.org/10.1002/app.28933

    Article  CAS  Google Scholar 

  34. Wang, W., Deng, L., Peng, Z., & Xiao, X. (2007). Study of the epoxydized magnetic hydroxyl particles as a carrier for immobilizing penicillin G acylase. Enzyme and Microbial Technology, 40(2), 255–261. https://doi.org/10.1016/j.enzmictec.2006.04.020

    Article  CAS  Google Scholar 

  35. Mateo, C., Palomo, J., Fernandez-Lorente, G., Guisan, J., & Fernandez-Lafuente, R. (2007). Improvement of enzyme activity, stability and selectivity via immobilization techniques. Enzyme and Microbial Technology, 40(6), 1451–1463. https://doi.org/10.1016/j.enzmictec.2007.01.018

    Article  CAS  Google Scholar 

  36. Bibi, Z., Shahid, F., Qader, S. A. U., & Aman, A. (2015). Agar-agar entrapment increases the stability of endo-β-1,4-xylanase for repeated biodegradation of xylan. International Journal of Biological Macromolecules, 75, 121–127. https://doi.org/10.1016/j.ijbiomac.2014.12.051

    Article  CAS  PubMed  Google Scholar 

  37. Kenneth, V., Esguerra, L., & Jean-Philip, L. (2017). A bioinspired catalytic aerobic functionalization of phenols: regioselective construction of aromatic C-N and C-O bonds. Catalysis, 7, 3477–3482.

    Google Scholar 

  38. Atacan, K., Çakıroğlu, B., & Özacar, M. (2017). Covalent immobilization of trypsin onto modified magnetite nanoparticles and its application for casein digestion. International Journal of Biological Macromolecules, 97, 148–155. https://doi.org/10.1016/j.ijbiomac.2017.01.023

    Article  CAS  PubMed  Google Scholar 

  39. Fernandez-Lopez, L., Rueda, N., Bartolome-Cabrero, R., Rodriguez, M., Albuquerque, T., Santos, J., & Fernandez-Lafuente, R. (2016). Improved immobilization and stabilization of lipase from Rhizomucor miehei on octyl-glyoxyl agarose beads by using CaCl2. Process Biochemistry, 51(1), 48–52. https://doi.org/10.1016/j.procbio.2015.11.015

    Article  CAS  Google Scholar 

  40. Zucca, P., & Sanjust, E. (2014). Inorganic materials as supports for covalent enzyme immobilization: methods and mechanisms. Molecules, 19(9), 14139–14194. https://doi.org/10.3390/molecules190914139

    Article  CAS  PubMed  Google Scholar 

  41. Srbová, J., Slováková, M., Křípalová, Z., Žárská, M., Špačková, M., Stránská, D., & Bílková, Z. (2016). Covalent biofunctionalization of chitosan nanofibers with trypsin for high enzyme stability. Reactive and Functional Polymers, 104, 38–44. https://doi.org/10.1016/j.reactfunctpolym.2016.05.009

    Article  CAS  Google Scholar 

  42. Hou, J., Dong, G., Ye, Y., & Chen, V. (2014). Laccase immobilization on titania nanoparticles and titania-functionalized membranes. Journal of Membrane Science, 452, 229–240. https://doi.org/10.1016/j.memsci.2013.10.019

    Article  CAS  Google Scholar 

  43. Song, J., Song, W., Yeo, S., Kim, H., & Lee, S. (2017). Covalent immobilization of enzyme on aminated woven poly (lactic acid) via ammonia plasma: evaluation of the optimum immobilization conditions. Textile Research Journal, 87(10), 1177–1191. https://doi.org/10.1177/0040517516648514

    Article  CAS  Google Scholar 

  44. Junoi, S., Chisti, Y., & Hansupalak, N. (2015). Optimal conditions for deproteinizing natural rubber using immobilized alkaline protease. Journal of Chemical Technology and Biotechnology, 90(1), 185–193. https://doi.org/10.1002/jctb.4304

    Article  CAS  Google Scholar 

  45. Hu, T., Cheng, J., Zhang, B., Lou, W., & Zong, M. (2015). Immobilization of alkaline protease on amino-functionalized magnetic nanoparticles and its efficient use for preparation of oat polypeptides. Industrial & Engineering Chemistry Research, 54(17), 4689–4698. https://doi.org/10.1021/ie504691j

    Article  CAS  Google Scholar 

  46. Wang, S., Zhang, C., Qi, B., Sui, X., Jiang, L., Li, Y., & Zhang, Q. (2014). Immobilized alcalase alkaline protease on the magnetic chitosan nanoparticles used for soy protein isolate hydrolysis. European Food Research and Technology, 239(6), 1051–1059. https://doi.org/10.1007/s00217-014-2301-1

    Article  CAS  Google Scholar 

  47. Gong, R., Zhang, J., Zhu, J., Wang, J., Lai, Q., & Jiang, B. (2013). Loofah sponge activated by periodate oxidation as a carrier for covalent immobilization of lipase. Korean Journal of Chemical Engineering, 30(8), 1620–1625. https://doi.org/10.1007/s11814-013-0102-z

    Article  CAS  Google Scholar 

  48. Nikolic, T., Milanovic, J., Kramar, A., Petronijevic, Z., Milenkovic, L., & Kostic, M. (2014). Preparation of cellulosic fibers with biological activity by immobilization of trypsin on periodate oxidized viscose fibers. Cellulose, 21(3), 1369–1380. https://doi.org/10.1007/s10570-014-0171-0

    Article  CAS  Google Scholar 

  49. An, N., Zhou, C., Zhang, X., Tong, D., & Yu, W. (2015). Immobilization of enzymes on clay minerals for biocatalysts and biosensors. Applied Clay Science, 114, 283–296. https://doi.org/10.1016/j.clay.2015.05.029

    Article  CAS  Google Scholar 

  50. Bibi, Z., Shahid, F., Qader, S., & Aman, A. (2015). Agar–agar entrapment increases the stability of endo-β-1, 4-xylanase for repeated biodegradation of xylan. International Journal of Biological Macromolecules, 75, 121–127. https://doi.org/10.1016/j.ijbiomac.2014.12.051

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

The work reported here was supported in part by the National Natural Science Foundation of China (21466022), international cooperation fund of the Ministry of Science and Technology (2014DFA61040), Jiangxi Province Key Science and Technology fund (20161BBF60057), and special fund for Jiangxi Province, thousands of people plan (GCXZ2014-124 100102102082).

Author information

Authors and Affiliations

  1. State Key Laboratory of Food Science and Technology, Engineering Research Center for Biomass Conversion, Ministry of Education, Nanchang University, Nanchang, 330047, China

    Lei-Peng Cao, Jing-Jing Wang, Ting Zhou, Roger Ruan & Yu-Huan Liu

  2. Center for Biorefining and Department of Bioproducts and Biosystems Engineering, University of Minnesota, Paul, MN, 55108, USA

    Roger Ruan

Authors
  1. Lei-Peng Cao
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jing-Jing Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ting Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Roger Ruan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yu-Huan Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Yu-Huan Liu.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Cao, LP., Wang, JJ., Zhou, T. et al. Bamboo (Phyllostachys pubescens) as a Natural Support for Neutral Protease Immobilization. Appl Biochem Biotechnol 186, 109–121 (2018). https://doi.org/10.1007/s12010-018-2697-3

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12010-018-2697-3

Keywords

Search

Navigation