Journal of Microbiology and Biotechnology

  • 제31권3호
  • /
  • Pages.419-428
  • /
  • 2021
  • /
  • 1017-7825(pISSN)
  • /
  • 1738-8872(eISSN)
한국미생물·생명공학회 (The Korean Society for Microbiology and Biotechnology)
권호 표지
DOI QR코드

DOI QR Code

Immobilization of GH78 α-L-Rhamnosidase from Thermotoga petrophilea with High-Temperature-Resistant Magnetic Particles Fe3O4-SiO2-NH2-Cellu-ZIF8 and Its Application in the Production of Prunin Form Naringin

  • Xu, Jin (Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University) ;
  • Shi, Xuejia (Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University) ;
  • Zhang, Xiaomeng (Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University) ;
  • Wang, Zhenzhong (Jiangsu Kanion Pharmaceutical Co., Ltd.) ;
  • Xiao, Wei (Jiangsu Kanion Pharmaceutical Co., Ltd.) ;
  • Zhao, Linguo (Jiangsu Co-Innovation Center of Efficient Processing and Utilization of Forest Resources, Nanjing Forestry University)
  • 투고 : 2020.04.24
  • 심사 : 2020.06.22
  • 발행 : 2021.03.28
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

To efficiently recycle GH78 thermostable rhamnosidase (TpeRha) and easily separate it from the reaction mixture and furtherly improve the enzyme properties, the magnetic particle Fe3O4-SiO2-NH2-Cellu-ZIF8 (FSNcZ8) was prepared by modifying Fe3O4-NH2 with tetraethyl silicate (TEOS), microcrystalline cellulose and zinc nitrate hexahydrate. FSNcZ8 displayed better magnetic stability and higher-temperature stability than unmodified Fe3O4-NH2 (FN), and it was used to adsorb and immobilize TpeRha from Thermotoga petrophilea 13995. As for properties, FSNcZ8-TpeRha showed optimal reaction temperature and pH of 90℃ and 5.0, while its highest activity approached 714 U/g. In addition, FSNcZ8-TpeRha had better higher-temperature stability than FN. After incubation at 80℃ for 3 h, the residual enzyme activities of FSNcZ8-TpeRha, FN-TpeRha and free enzyme were 93.5%, 63.32%, and 62.77%, respectively. The organic solvent tolerance and the monosaccharides tolerance of FSNcZ8-TpeRha, compared with free TpeRha, were greatly improved. Using naringin (1 mmol/l) as the substrate, the optimal conversion conditions were as follows: FSNcZ8-TpeRha concentration was 6 U/ml; induction temperature was 80℃; the pH was 5.5; induction time was 30 min, and the yield of products was the same as free enzyme. After repeating the reaction 10 times, the conversion of naringin remained above 80%, showing great improvement of the catalytic efficiency and repeated utilization of the immobilized α-L-rhamnosidase.

키워드

참고문헌

  1. Lim T, Jung H, Hwang KT. 2015. Bioconversion of cyanidin-3-Rutinoside to Cyanidin-3-Glucoside in black raspberry by crude α-L-Rhamnosidase from Aspergillus Species. J. Microbiol. Biotechnol. 25: 1842-1848. https://doi.org/10.4014/jmb.1503.03098
  2. Karel DW, Daniela II, Bram S, Lenka W, Helena P, Wim S. 2013. Chemoenzymatic synthesis of α-L-rhamnosides using recombinant α-L-rhamnosidase from Aspergillus terreus, Bioresour. Technol. 147: 640-644. https://doi.org/10.1016/j.biortech.2013.08.083
  3. Markosova K, Weignerova L, Rosenberg M, Kren V, Rebros M. 2015. Upscale of recombinant α-L-rhamnosidase production by Pichia pastoris Mut(S) strain. Front. Microbiol. 6: 1140-1150.
  4. Yadav V , Yadav PK , Yadav S , Yadav KDS. 2010. α-L-rhamnosidase: a review. Process Biochem. 45: 1226-1235. https://doi.org/10.1016/j.procbio.2010.05.025
  5. Zhang R, Zhang B, Li G, Xie T, Hu T, Luo Z. 2015. Enhancement of ginsenoside Rg(1) in Panax ginseng hairy root by overexpressing the α-L-rhamnosidase gene from Bifidobacterium breve. Biotechnol. Lett. 37: 2091-2096. https://doi.org/10.1007/s10529-015-1889-y
  6. Zhang R, Zhang B, Xie T, Li G, Tuo Y, Xiang Y. 2015. Biotransformation of rutin to isoquercitrin using recombinant rhamnosidase from Bifidobacterium breve. Biotechnol. Lett. 37: 1257-1264. https://doi.org/10.1007/s10529-015-1792-6
  7. Weignerova L, Marhol P, Kren V. 2012. Preparatory production of quercetin-3-β-D-glucopyranoside using alkali-tolerant thermostable α-L-rhamnosidase from Aspergillus terreus. Bioresour. Technol. 115: 222-227. https://doi.org/10.1016/j.biortech.2011.08.029
  8. Ge L, Li D, Wu T, Zhao LG, DING G, WANG ZZ, et al. 2018.B-factor-saturation mutagenesis as a strategy to increase the thermostability of α-L-rhamnosidase from Aspergillus terreus. J. Biotechnol. 275: 17-23. https://doi.org/10.1016/j.jbiotec.2018.03.013
  9. Zverlov VV , Hertel C , Bronnenmeier K , Hroch A, Kellermann J, Schwarz WH. 2010. The thermostable α-L-rhamnosidase RamA of Clostridium stercorarium: biochemical characterization and primary structure of a bacterial α-L-rhamnosidase hydrolase, a new type of inverting glycoside hydrolase. Mol. Microbiol. 35: 173-179. https://doi.org/10.1046/j.1365-2958.2000.01691.x
  10. Li JR, Kuppler RJ, Zhou HC. 2009. Selective gas adsorption and separation in metal-organic frameworks. Chem. Soc. Rev. 38: 1477-1504. https://doi.org/10.1039/b802426j
  11. Yng M, Jiang H, Ning C, Wei D, Su E. 2018. Preparation of combined cross-linked enzyme aggregates and its application in synthesis of chiral alcohols by the asymmetric reduction of carbanyl group. Chem. J. Chinese Universities 39: 54-63.
  12. Zhang S , Zhang Y , Liu J , Xu Q, Xiao H, Wang X, et al. 2013. Thiol modified Fe3O4@SiO2 as a robust, high effective, and recycling magnetic sorbent for mercury removal. Chem. Eng. J. 226: 30-38. https://doi.org/10.1016/j.cej.2013.04.060
  13. Shao Y, Jing T, Tian J, Zheng Y. 2015. Graphene oxide-based Fe3O4 nanoparticles as a novel scaffold for the immobilization of porcine pancreatic lipase. Rsc. Adv. 5: 103943-103955. https://doi.org/10.1039/C5RA19276E
  14. Wu JC, Hutchings CH, Lindsay MJ, Werner CJ, Bundy BC. 2015. Enhanced enzyme stability through site-directed covalent immobilization. J. Biotechnol. 193: 83-90. https://doi.org/10.1016/j.jbiotec.2014.10.039
  15. Shi X, Zhao L, Pei J, Ge L, Wan P, Wang Z, et al. 2018. Highly enhancing the characteristics of immobilized thermostable β-glucosidase by Zn2+. Process Biochem. 66: 89-96. https://doi.org/10.1016/j.procbio.2018.01.004
  16. Wu X, Ge J, Yang C, Hou M, Liu Z. 2015. Facile synthesis of multiple enzyme-containing metal-organic frameworks in a biomolecule-friendly environment. Chem. Commun. 51: 13408-13411. https://doi.org/10.1039/C5CC05136C
  17. Cao SL, Xu H, Lai LH, Gu WM, Xu P, Xiong J, et al. 2017. Magnetic ZIF-8/cellulose/Fe3O4 nanocomposite: preparation, characterization, and enzyme immobilization. Bioresour. Bioprocess. 4: 56-63. https://doi.org/10.1186/s40643-017-0186-0
  18. Wu X, Yang C, Ge J. 2017. Green synthesis of enzyme/metal-organic framework composites with high stability in protein denaturing solvents. Bioresour. Bioprocess. 4: 24-32. https://doi.org/10.1186/s40643-017-0154-8
  19. Liu Z, Wang H, Liu C, Jiang Y, Yu G, Mu X. 2012. Magnetic cellulose-chitosan hydrogels prepared from ionic liquids as reusable adsorbent for removal of heavy metal ions. Chem. Commun. 48: 7350-7352. https://doi.org/10.1039/c2cc17795a
  20. Wehner T, Mandel K, Schneider M, Sextl G, Muellerbuschbaum K. 2016. Superparamagnetic luminescent MOF@Fe3O4/SiO2 composite particles for signal augmentation by magnetic harvesting as potential water detectors. Acs Appl. Mater. Interfaces 8: 5445-5452. https://doi.org/10.1021/acsami.5b11965
  21. Ali Z, Tian L, Zhang B, Ali N, Khan M, Zhang Q. 2017. Synthesis of paramagnetic dendritic silica nanomaterials with fibrous pore structure (Fe3O4@KCC-1) and their application in immobilization of lipase from Candida rugosa with enhanced catalytic activity and stability. New J. Chem. 41: 8222-8232. https://doi.org/10.1039/C7NJ01912B
  22. Xie J, Zhang S, Tong X, Wu T, Pei J, Zhao L. 2020. Biochemical characterization of a novel hyperthermophilic α-L-Rhamnosidase from Thermotoga petrophila and its application in production of icaritin from epimedin C with a thermostable β-glucosidase. Process Biochem. 93: 115-124. https://doi.org/10.1016/j.procbio.2020.03.019
  23. Xie J, Zhao D, Zhao L, Pei J, Wei X, Gang D, et al. 2015. Overexpression and characterization of a Ca2+ activated thermostable β-glucosidase with high ginsenoside Rb1 to ginsenoside 20(S)-Rg3 bioconversion productivity. J. Ind. Microbiol. Biotechnol. 42: 839-850. https://doi.org/10.1007/s10295-015-1608-7
  24. Wu X, Yang C, Ge J. 2017. Green synthesis of enzyme/metal-organic framework composites with high stability in protein denaturing solvents. Bioresour. Bioprocess. 4: 24-32. https://doi.org/10.1186/s40643-017-0154-8
  25. Lyu F, Zhang Y, Zare RN, Ge J, Liu Z. 2014. One-pot synthesis of protein-embedded metal-organic frameworks with enhanced biological activities, Nano Lett. 14: 5761-5765. https://doi.org/10.1021/nl5026419
  26. Su E, Meng Y, Ning C, Ma X, Deng S. 2018. Magnetic combined cross-linked enzyme aggregates (Combi-CLEAs) for cofactor regeneration in the synthesis of chiral alcohol. J. Biotechnol. 271: 1-7. https://doi.org/10.1016/j.jbiotec.2018.02.007
  27. Talekar S, Ghodake V, Ghotage T, Rathod P, Deshmukh P, Nadar S, et al. 2012. Novel magnetic cross-linked enzyme aggregates (magnetic CLEAs) of α-amylase, Bioresour. Technol. 123: 542-547. https://doi.org/10.1016/j.biortech.2012.07.044
  28. Grewal J, Ahmad R, Khare SK. 2017. Development of cellulase-nanoconjugates with enhanced ionic liquid and thermal stability for in situ lignocellulose saccharification. Bioresour. Technol. 224: 236-243. https://doi.org/10.1016/j.biortech.2016.11.002
  29. Rios NS , Pinheiro MP, dos Santos Jose Cleiton S, Fonseca TDS, Lima LD, Mattos MC de, et al. 2016. Strategies of covalent immobilization of a recombinant Candida antarctica lipase B on pore-expanded SBA-15 and its application in the kinetic resolution of (R,S)-Phenylethyl acetate. J. Mol. Catal B: Enzym. 133: 246-258. https://doi.org/10.1016/j.molcatb.2016.08.009
  30. Grewal J, Ahmad R, Khare SK. 2017. Development of cellulase-nanoconjugates with enhanced ionic liquid and thermal stability for, in situ, lignocellulose saccharification. Bioresour. Technol. 242: 236-343. https://doi.org/10.1016/j.biortech.2017.04.007
  31. Wu R, Dong QH, Sun Y, Su EZ. 2019. Efficient enzyme immobilization by combining adsorption and cellulose membrane coating. Chem. J. Chinese Univ. 40: 1888-1896.
  32. Zhai Z, Yang T, Zhang B, Zhang J. 2015. Effects of metal ions on the catalytic degradation of dicofol by cellulase. J. Environ. Sci. 33: 163-168. https://doi.org/10.1016/j.jes.2014.12.023
  33. Zhao R, Hu M, Li S, Zhai Q, Jiang Y. 2017. Immobilization of chloroperoxidase in metal organic framework and its catalytic performance. Acta Chimcal Sinica 75: 293-299. https://doi.org/10.6023/A16110593
  34. Fennouh S, Casimiri V, Geloso-Meyer A, Burstein C. 1998. Kinetic study of heavy metal salt effects on the activity of L-lactate dehydrogenase in solution or immobilized on an oxygen electrode. Biosens. Biolectron. 13: 903-909. https://doi.org/10.1016/S0956-5663(98)00062-1
  35. Mazzaferro LS, Orrillo GA, Ledesma P, Breccia JD. 2008. Dose-dependent significance of monosaccharides on intracellular α-L-rhamnosidase activity from Pseudoalteromonas sp. Biotechnol. Lett. 30: 2147-2150. https://doi.org/10.1007/s10529-008-9810-6