Utilization of one novel deep-sea microbial protease sin3406-1 in the preparation of ethyl (S)-3-hydroxybutyrate through kinetic resolution

  • Jinlong Huang
  • Yongkai Xu
  • Yun Zhang
  • Aijun Sun
  • Yunfeng Hu
Original Paper


One novel protease sin3406-1 was identified from Streptomyces niveus SCSIO 3406, which was isolated from the deep sea of the South China Sea, and heterologously expressed in E. coli BL21(DE3). Protease sin3406-1 was further used as a green biocatalyst in the kinetic resolution of racemic ethyl-3-hydroxybutyrate. After careful process optimization, chiral product ethyl (S)-3-hydroxybutyrate was generated with an enantiomeric excess of over 99% and a conversion rate of up to 50% through direct hydrolysis of inexpensive racemic ethyl-3-hydroxybutyrate catalyzed by sin3406-1. Interestingly, protease sin3406-1 exhibited the same enantio-preference as that of esterase PHE21 during the asymmetric hydrolysis of the ester bonds of racemic ethyl-3-hydroxybutyrate. Through mutation studies and molecular docking, we also demonstrated that the four residues close to the catalytic center, S85, A86, Q87 and Y254, played key roles in both the hydrolytic activity and the enantioselectivity of protease sin3406-1, possibly through forming hydrogen bonds between the enzyme and the substrates. Deep-sea microbial proteases represented by sin3406-1 are new contributions to the biocatalyst library for the preparation of valuable chiral drug intermediates and chemicals through enzymatic kinetic resolution.


Biocatalysis Marine microorganisms Novel protease Kinetic resolution Ethyl (S)-3-hydroxybutyrate 



We are grateful for the financial supports from Scientific and Technological Project of Ocean and Fishery from Guangdong Province (A201701C12), the Priority Research Program of the Chinese Academy of Sciences (XDA11030404) and Guangzhou Science and Technology Plan Projects (201510010012).

Compliance with ethical standards

Conflict of interest

The authors all declare that they have no conflict of interest.

Supplementary material

11274_2018_2513_MOESM1_ESM.docx (5.8 mb)
Supplementary material 1 (DOCX 5897 KB)


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Jinlong Huang
    • 1
    • 2
    • 3
  • Yongkai Xu
    • 4
  • Yun Zhang
    • 1
    • 2
  • Aijun Sun
    • 1
    • 2
  • Yunfeng Hu
    • 1
    • 2
    • 5
  1. 1.CAS Key Laboratory of Tropical Marine Bio-resources and Ecology, South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhouPeople’s Republic of China
  2. 2.Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of OceanologyChinese Academy of SciencesGuangzhouPeople’s Republic of China
  3. 3.College of Life ScienceGuangxi Normal UniversityGuilinPeople’s Republic of China
  4. 4.Affiliated Hospital of Shandong University of Traditional Chinese MedicineJinanPeople’s Republic of China
  5. 5.South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation CenterGuangzhouPeople’s Republic of China

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