机构:[1]CAS Key Laboratory of Tropical Marine Bio‑resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, People’s Republic of China [2]Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, People’s Republic of China [3]College of Life Science, Guangxi Normal University, Guilin 541004, People’s Republic of China [4]Affiliated Hospital of Shandong University of Traditional Chinese Medicine, Jinan 250014, Shandong, People’s Republic of China [5]South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Guangzhou, People’s Republic of China
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.
基金:
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).
语种:
外文
PubmedID:
中科院(CAS)分区:
出版当年[2017]版:
大类|3 区工程技术
小类|4 区生物工程与应用微生物
最新[2025]版:
大类|3 区生物学
小类|3 区生物工程与应用微生物
第一作者:
第一作者机构:[1]CAS Key Laboratory of Tropical Marine Bio‑resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, People’s Republic of China [2]Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, People’s Republic of China [3]College of Life Science, Guangxi Normal University, Guilin 541004, People’s Republic of China
通讯作者:
通讯机构:[1]CAS Key Laboratory of Tropical Marine Bio‑resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, People’s Republic of China [2]Guangdong Key Laboratory of Marine Materia Medica, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510301, People’s Republic of China [5]South China Sea Bio-Resource Exploitation and Utilization Collaborative Innovation Center, Guangzhou, People’s Republic of China
推荐引用方式(GB/T 7714):
Huang Jinlong,Xu Yongkai,Zhang Yun,et al.Utilization of one novel deep-sea microbial protease sin3406-1 in the preparation of ethyl (S)-3-hydroxybutyrate through kinetic resolution.[J].World journal of microbiology & biotechnology.2018,34(9):124.doi:10.1007/s11274-018-2513-9.
APA:
Huang Jinlong,Xu Yongkai,Zhang Yun,Sun Aijun&Hu Yunfeng.(2018).Utilization of one novel deep-sea microbial protease sin3406-1 in the preparation of ethyl (S)-3-hydroxybutyrate through kinetic resolution..World journal of microbiology & biotechnology,34,(9)
MLA:
Huang Jinlong,et al."Utilization of one novel deep-sea microbial protease sin3406-1 in the preparation of ethyl (S)-3-hydroxybutyrate through kinetic resolution.".World journal of microbiology & biotechnology 34..9(2018):124