Advertisement

Rational design of engineered microbial cell surface multi-enzyme co-display system for sustainable NADH regeneration from low-cost biomass

  • Lei Han
  • Bo Liang
  • Jianxia Song
Biocatalysis - Original Paper

Abstract

As an important cofactor, NADH is essential for most redox reactions and biofuel cells. However, supply of exogenous NADH is challenged, due to the low production efficiency and high cost of NADH regeneration system, as well as low stability of NADH. Here, we constructed a novel cell surface multi-enzyme co-display system with ratio- and space-controllable manner as exogenous NADH regeneration system for the sustainable NADH production from low-cost biomass. Dockerin-fused glucoamylase (GA) and glucose dehydrogenase (GDH) were expressed and assembled on the engineered bacterial surfaces, which displayed protein scaffolds with various combinations of different cohesins. When the ratio of GA and GDH was 3:1, the NADH production rate of the whole-cell biocatalyst reached the highest level using starch as substrate, which was three times higher than that of mixture of free enzymes, indicating that the highly ordered spatial organization of enzymes would promote reactions, due to the ratio of enzymes and proximity effect. To confirm performance of the established NADH regeneration system, the highly efficient synthesis of l-lactic acid (l-LA) was conducted by the system and the yield of l-LA (16 g/L) was twice higher than that of the mixture of free enzymes. The multi-enzyme co-display system showed good stability in the cyclic utilization. In conclusion, the novel sustainable NADH system would provide a cost-effective strategy to regenerate cofactor from low-cost biomass.

Keywords

Microbial cell surface display technology Multi-enzyme co-display system Cofactor regeneration system Low-cost biomass Lactic acid 

Notes

Acknowledgements

The authors are grateful for helpful discussion of Prof. Yingang Feng in Qingdao institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences. This work was supported by National Natural Science Foundation of China (Grant No. 21705087) and Distinguished Scholars of Qingdao Agricultural University (No. 6631117015).

References

  1. 1.
    Bugg TDH, Rahmanpour R (2015) Enzymatic conversion of lignin into renewable chemicals. Curr Opin Chem Biol 29:10–17.  https://doi.org/10.1016/j.cbpa.2015.06.009 CrossRefPubMedGoogle Scholar
  2. 2.
    Cui ZM, Zhang JD, Fan XJ et al (2017) Highly efficient bioreduction of 2-hydroxyacetophenone to (S)- and (R)-1-phenyl-1,2-ethanediol by two substrate tolerance carbonyl reductases with cofactor regeneration. J Biotechnol 243:1–9.  https://doi.org/10.1016/j.jbiotec.2016.12.016 CrossRefPubMedGoogle Scholar
  3. 3.
    Dueber JE, Wu GC, Malmirchegini GR et al (2009) Synthetic protein scaffolds provide modular control over metabolic flux. Nat Biotechnol 27:753–759.  https://doi.org/10.1038/nbt.1557 CrossRefPubMedGoogle Scholar
  4. 4.
    Fan LH, Zhang ZJ, Yu XY et al (2012) Self-surface assembly of cellulosomes with two miniscaffoldins on Saccharomyces cerevisiae for cellulosic ethanol production. Proc Natl Acad Sci USA 109:13260–13265.  https://doi.org/10.1073/pnas.1209856109 CrossRefPubMedGoogle Scholar
  5. 5.
    Goacher RE, Selig MJ, Master ER (2014) Advancing lignocellulose bioconversion through direct assessment of enzyme action on insoluble substrates. Curr Opin Biotechnol 27:123–133.  https://doi.org/10.1016/j.copbio.2014.01.009 CrossRefPubMedGoogle Scholar
  6. 6.
    Haimovitz R, Barak Y, Morag E et al (2008) Cohesin-dockerin microarray: diverse specificities between two complementary families of interacting protein modules. Proteomics 8:968–979.  https://doi.org/10.1002/pmic.200700486 CrossRefPubMedGoogle Scholar
  7. 7.
    Han L, Liu A (2017) Novel cell–inorganic hybrid catalytic interfaces with enhanced enzymatic activity and stability for sensitive biosensing of paraoxon. ACS Appl Mater Interfaces 9:6894–6901.  https://doi.org/10.1021/acsami.6b15992 CrossRefPubMedGoogle Scholar
  8. 8.
    Han L, Zhao Y, Cui S et al (2017) Redesigning of microbial cell surface and its application to whole-cell biocatalysis and biosensors. Appl Biochem Biotechnol.  https://doi.org/10.1007/s12010-017-2662-6 CrossRefPubMedGoogle Scholar
  9. 9.
    Hummel W, Groger H (2014) Strategies for regeneration of nicotinamide coenzymes emphasizing self-sufficient closed-loop recycling systems. J Biotechnol 191:22–31.  https://doi.org/10.1016/j.jbiotec.2014.07.449 CrossRefPubMedGoogle Scholar
  10. 10.
    Hyeon JE, Kim SW, Park C et al (2015) Efficient biological conversion of carbon monoxide (CO) to carbon dioxide (CO2) and for utilization in bioplastic production by Ralstonia eutropha through the display of an enzyme complex on the cell surface. Chem Commun 51:10202–10205.  https://doi.org/10.1039/c5cc00832h CrossRefGoogle Scholar
  11. 11.
    Ishii J, Okazaki F, Djohan AC et al (2016) From mannan to bioethanol: cell surface co-display of beta-mannanase and beta-mannosidase on yeast Saccharomyces cerevisiae. Biotechnol Biofuels 9:188.  https://doi.org/10.1186/S13068-016-0600-4 CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Kam Y, Sung M, Cho H et al (2017) Utilization of starch-enriched brewery (rice wine) waste for mixotrophic cultivation of Ettlia sp. YC001 used in biodiesel production. Appl Biochem Biotechnol 183:1478–1487.  https://doi.org/10.1007/s12010-017-2515-3 CrossRefPubMedGoogle Scholar
  13. 13.
    Khatun MM, Liu CG, Zhao XQ et al (2017) Consolidated ethanol production from Jerusalem artichoke tubers at elevated temperature by Saccharomyces cerevisiae engineered with inulinase expression through cell surface display. J Ind Microbiol Biot 44:295–301.  https://doi.org/10.1007/s10295-016-1881-0 CrossRefGoogle Scholar
  14. 14.
    Kowalsky CA, Whitehead TA (2016) Determination of binding affinity upon mutation for type I dockerin-cohesin complexes from Clostridium thermocellum and Clostridium cellulolyticum using deep sequencing. Proteins 84:1914–1928.  https://doi.org/10.1002/prot.25175 CrossRefPubMedGoogle Scholar
  15. 15.
    Lee SH, Choi JH, Park SH et al (2004) Enantioselective resolution of racemic compounds by cell surface displayed lipase. Enzyme Microb Technol 35:429–436.  https://doi.org/10.1016/j.enzmictec.2004.06.005 CrossRefGoogle Scholar
  16. 16.
    Liu F, Banta S, Chen W (2013) Functional assembly of a multi-enzyme methanol oxidation cascade on a surface-displayed trifunctional scaffold for enhanced NADH production. Chem Commun 49:3766–3768.  https://doi.org/10.1039/c3cc40454d CrossRefGoogle Scholar
  17. 17.
    Liu WF, Wang P (2007) Cofactor regeneration for sustainable enzymatic biosynthesis. Biotechnol Adv 25:369–384.  https://doi.org/10.1016/j.biotechadv.2007.03.002 CrossRefPubMedGoogle Scholar
  18. 18.
    Liu Z, Inokuma K, Ho SH et al (2015) Combined cell-surface display- and secretion-based strategies for production of cellulosic ethanol with Saccharomyces cerevisiae. Biotechnol Biofuels 8:162.  https://doi.org/10.1186/S13068-015-0344-6 CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Martinez FAC, Balciunas EM, Salgado JM et al (2013) Lactic acid properties, applications and production: a review. Trends Food Sci Technol 30:70–83.  https://doi.org/10.1016/j.tifs.2012.11.007 CrossRefGoogle Scholar
  20. 20.
    Munoz-Gutierrez I, Oropeza R, Gosset G et al (2012) Cell surface display of a beta-glucosidase employing the type V secretion system on ethanologenic Escherichia coli for the fermentation of cellobiose to ethanol. J Ind Microbiol Biotechnol 39:1141–1152.  https://doi.org/10.1007/s10295-012-1122-0 CrossRefPubMedGoogle Scholar
  21. 21.
    Nair RB, Kabir MM, Lennartsson PR et al (2017) Integrated process for ethanol, biogas, and edible filamentous fungi-based animal feed production from dilute phosphoric acid-pretreated wheat straw. Appl Biochem Biotechnol.  https://doi.org/10.1007/s12010-017-2525-1 PubMedCentralCrossRefPubMedGoogle Scholar
  22. 22.
    Pages S, Belaich A, Belaich JP et al. (1997) Species-specificity of the cohesin–dockerin interaction between Clostridium thermocellum and Clostridium cellulolyticum: prediction of specificity determinants of the dockerin domain. Proteins 29:517–527. https://doi.org/10.1002/(sici)1097-0134(199712)29:4<517::aid-prot11>3.3.co;2-iGoogle Scholar
  23. 23.
    Pongtharangkul T, Chuekitkumchorn P, Suwanampa N et al (2015) Kinetic properties and stability of glucose dehydrogenase from Bacillus amyloliquefaciens SB5 and its potential for cofactor regeneration. Amb Express 5:68.  https://doi.org/10.1186/s13568-015-0157-9 CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Schoffelen S, Van Hest JCM (2012) Multi-enzyme systems: bringing enzymes together in vitro. Soft Matter 8:1736–1746.  https://doi.org/10.1039/c1sm06452e CrossRefGoogle Scholar
  25. 25.
    Tsai SL, Oh J, Singh S et al (2009) Functional assembly of minicellulosomes on the Saccharomyces cerevisiae cell surface for cellulose hydrolysis and ethanol production. Appl Environ Microbiol 75:6087–6093.  https://doi.org/10.1128/Aem.01538-09 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Ueda M (2016) Establishment of cell surface engineering and its development. Biosci Biotechnol Biochem 80:1243–1253.  https://doi.org/10.1080/09168451.2016.1153953 CrossRefPubMedGoogle Scholar
  27. 27.
    Van Bloois E, Winter RT, Kolmar H et al (2011) Decorating microbes: surface display of proteins on Escherichia coli. Trends Biotechnol 29:79–86.  https://doi.org/10.1016/j.tibtech.2010.11.003 CrossRefPubMedGoogle Scholar
  28. 28.
    Wang L, Cai Y, Zhu L et al (2014) Major role of NAD-dependent dehydrogenases in the production of l-lactic acid with high optical purity by the thermophile Bacillus coagulans. Appl Environ Microbiol 80:7134–7141.  https://doi.org/10.1128/AEM.01864-14 CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Wieczorek AS, Martin VJJ (2012) Effects of synthetic cohesin-containing scaffold protein architecture on binding dockerin-enzyme fusions on the surface of Lactococcus lactis. Microb Cell Fact 11:160.  https://doi.org/10.1186/1475-2859-11-160 CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Zhang M, Xie L, Yin Z et al (2016) Biorefinery approach for cassava-based industrial wastes: current status and opportunities. Bioresour Technol 215:50–62.  https://doi.org/10.1016/j.biortech.2016.04.026 CrossRefPubMedGoogle Scholar
  31. 31.
    Zhao H, Van der Donk WA (2003) Regeneration of cofactors for use in biocatalysis. Curr Opin Biotechnol 14:583–589.  https://doi.org/10.1016/j.copbio.2003.09.007 CrossRefPubMedGoogle Scholar

Copyright information

© Society for Industrial Microbiology and Biotechnology 2018

Authors and Affiliations

  1. 1.College of Chemistry and Pharmaceutical SciencesQingdao Agricultural UniversityQingdaoChina
  2. 2.Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoChina
  3. 3.University of Chinese Academy of SciencesBeijingChina

Personalised recommendations