Biotechnology Letters

, Volume 41, Issue 11, pp 1333–1341 | Cite as

A kinetic model to optimize and direct the dose ratio of Dsz enzymes in the 4S desulfurization pathway in vitro and in vivo

  • Lu Li
  • Lei Ye
  • Zhijie Guo
  • Wei Zhang
  • Xihao Liao
  • Ying LinEmail author
  • Shuli LiangEmail author
Original Research Paper



To enhance the biodesulfurization rate using a kinetic model that directs the ratio of Dsz enzymes.


This study established a kinetic model that predicted the optimal ratio of Dsz enzymes in the 4S biodesulfurization system to be A:B:C = 1:2:4 and 1:4:2. When BCAD+1A+4B+2C, the conversion rate of dibenzothiophene (DBT) to 2-hydroxybiphenyl (HBP) was close to 100% in vitro. When the gene dose of dszC was increased, the HBP yield of the recombinant strain BL21(DE3)/BCAD + C reached approximately 0.012 mM in vivo, which was approximately 6-fold higher than that of the BCAD strain.


According to the results predicted by the enzyme kinetic model, maintaining higher concentrations of DszC and DszB in the desulfurization system can effectively improve the desulfurization efficiency.


Biodesulfurization Kinetic model Dsz enzyme ratio Gene dosagen 











Dibenzothiophene sulfone


2-(2-Hydroxybiphenyl) 2 sulfinic acid salt


High-performance liquid chromatography


Supporting information

Supplementary Figure 1. The rate of conversion of DBT to HBP by BL21(DE3)/BADC+C.


This research was financially supported by the National Natural Science Foundation of China (Grant No. 31470159), the National Science Foundation for Young Scientists of China (Grant No. 31400062).

Compliance with ethical standards

Conflict of interest

The authors declare no competing financial interest.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

10529_2019_2730_MOESM1_ESM.docx (175 kb)
Supplementary file1 (DOCX 175 kb)


  1. Abinfuentes A, Mohamed ES, Wang DIC, Prather KLJ (2013) Exploring the mechanism of biocatalyst inhibition in microbial desulfurization. Appl Environ Microbiol 79:7807–7817CrossRefGoogle Scholar
  2. Agarwal M, Dikshit PK, Bhasarkar JB, Borah AJ, Moholkar VS (2016) Physical insight into ultrasound-assisted biodesulfurization using free and immobilized cells of Rhodococcus rhodochrous MTCC 3552. Chem Eng J 295:254–267CrossRefGoogle Scholar
  3. Calzada J, Heras S, Alcon A, Santos VE, Garciaochoa F (2009) Biodesulfurization of dibenzothiophene (DBT) using Pseudomonas putida CECT 5279: a biocatalyst formulation comparison. Energy Fuels 23:155–166CrossRefGoogle Scholar
  4. Dejaloud A, Vahabzadeh F, Habibi A (2017) Ralstonia eutropha as a biocatalyst for desulfurization of dibenzothiophene. Bioprocess Biosyst Eng 40:1–12CrossRefGoogle Scholar
  5. Denome SA, Olson ES, Young KD (1993) Identification and cloning of genes involved in specific desulfurization of dibenzothiophene by Rhodococcus sp. strain IGTS8. Appl Environ Microbiol 59:2837–2843PubMedPubMedCentralGoogle Scholar
  6. Ferreira P, Sousa SF, Fernandes PA, Ramos MJ (2017) Improving the catalytic power of the DszD enzyme for the biodesulfurization of crude oil and derivatives. Chemistry 23:17231–17241CrossRefPubMedGoogle Scholar
  7. Galán B, Díaz E, García JL (2000) Enhancing desulphurization by engineering a flavin reductase-encoding gene cassette in recombinant biocatalysts. Environ Microbiol 2:687–694CrossRefPubMedGoogle Scholar
  8. Gumpf JJ, Idrizi K, Watkins L (2017) Expression, purification, and characterization of codon optimized and mutant variations of DszB from N. asteroides. FASEB J 31:764–767Google Scholar
  9. Li L et al (2019) Improved efficiency of the desulfurization of oil sulfur compounds in Escherichia coli using a combination of desensitization engineering and DszC overexpression. ACS Synth Biol 8:1441–1451CrossRefPubMedGoogle Scholar
  10. Martínez I, Mohamed ES, Rozas D, García JL, Díaz E (2016) Engineering synthetic bacterial consortia for enhanced desulfurization and revalorization of oil sulfur compounds. Metab Eng 35:46CrossRefPubMedGoogle Scholar
  11. Martínez I, Elsaid MM, Santos VE, García JL, Garcíaochoa F, Díaz E (2017) Metabolic and process engineering for biodesulfurization in gram-negative bacteria. J Biotechnol 262:47–55CrossRefPubMedGoogle Scholar
  12. Rollin JA et al (2015) High-yield hydrogen production from biomass by in vitro metabolic engineering: mixed sugars coutilization and kinetic modeling. Proc Natl Acad Sci USA 112:4964–4969CrossRefPubMedGoogle Scholar
  13. Santos VE, AlcãN A, Martã­N AB, GãMez E, Garcia-Ochoa F, (2007) Desulfurization of dibenzothiophene using the 4S enzymatic route: influence of operational conditions on initial reaction rates. Biocatalysis 25:286–294CrossRefGoogle Scholar
  14. Yu Y, Fursule IA, Mills LC, Englert DL, Berron BJ, Payne CM (2017) CHARMM force field parameters for 2′-hydroxybiphenyl-2-sulfinate, 2-hydroxybiphenyl, and related analogs. J Mol Graph Model 72:32–42CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Guangdong Key Laboratory of Fermentation and Enzyme Engineering, School of Biology and Biological EngineeringSouth China University of TechnologyGuangzhouChina
  2. 2.Guangdong Research Center of Industrial Enzyme and Green Manufacturing Technology, School of Biology and Biological EngineeringSouth China University of TechnologyGuangzhouChina
  3. 3.College of Science and EngineeringJinan UniversityGuangzhouChina

Personalised recommendations