PMID- 31240368
OWN - NLM
STAT- MEDLINE
DCOM- 20191230
LR  - 20191230
IS  - 1432-0614 (Electronic)
IS  - 0175-7598 (Linking)
VI  - 103
IP  - 16
DP  - 2019 Aug
TI  - Identification and characterization of a meta-cleavage product hydrolase involved 
      in biphenyl degradation from Arthrobacter sp. YC-RL1.
PG  - 6825-6836
LID - 10.1007/s00253-019-09956-z [doi]
AB  - Polychlorinated biphenyls (PCBs) are a group of persistent organic pollutants 
      (POPs) widely existing in the environment. Arthrobacter sp. YC-RL1 is a 
      biphenyl-degrading bacterium that shows metabolic versatility towards aromatic 
      compounds. A 2-hydroxy-6-oxo-6-phenylhexa-2, 4-dienoate (HOPDA) hydrolase (BphD) 
      gene involved in the biodegradation of biphenyl was cloned from strain YC-RL1 and 
      heterologously expressed in Escherichia coli BL21 (DE3). The recombinant 
      BphD(YC-RL1) was purified and characterized. BphD(YC-RL1) showed the highest 
      activity at 45  degrees C and pH 7. It was stable under a wide range of temperature 
      (20-50  degrees C). The enzyme had a K(m) value of 0.14 mM, K(cat) of 11.61 s(-1), and 
      V(max) of 0.027 U/mg. Temperature dependence catalysis exhibited a biphasic 
      Arrhenius Plot with a transition at 20  degrees C. BphD(YC-RL1) was inactivated by SDS, 
      Tween 20, Tween 80, Trition X-100, DTT, CHAPS, NBS, PMSF, and DEPC, but 
      insensitive to EDTA. Site-directed mutagenesis of the active-site residues 
      revealed that the catalytic triad residues (Ser115, His275, and Asp247) of 
      BphD(YC-RL1) were necessary for its activity. The investigation of BphD(YC-RL1) 
      not only provides new potential enzyme resource for the biodegradation of 
      biphenyl but also helps deepen our understanding on the catalytic process and 
      mechanism.
FAU - Jia, Yang
AU  - Jia Y
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China.
FAU - Wang, Junhuan
AU  - Wang J
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China.
FAU - Ren, Chao
AU  - Ren C
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China.
FAU - Nahurira, Ruth
AU  - Nahurira R
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China.
FAU - Khokhar, Ibatsam
AU  - Khokhar I
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China.
FAU - Wang, Jiayi
AU  - Wang J
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China.
FAU - Fan, Shuanghu
AU  - Fan S
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China.
FAU - Yan, Yanchun
AU  - Yan Y
AD  - Graduate School, Chinese Academy of Agricultural Sciences, Beijing, 100081, 
      China. yanyanchun@caas.cn.
LA  - eng
GR  - 31540067/National Natural Science Foundation of China (CN)/
GR  - 21876201/National Natural Science Foundation of China/
GR  - 1610042017001/Basic Research Fund of CAAS/
GR  - 1610042018005/Basic Research Fund of CAAS/
GR  - 1610042018006/Basic Research Fund of CAAS/
PT  - Journal Article
DEP - 20190626
PL  - Germany
TA  - Appl Microbiol Biotechnol
JT  - Applied microbiology and biotechnology
JID - 8406612
RN  - 0 (Biphenyl Compounds)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Fungicides, Industrial)
RN  - 2L9GJK6MGN (diphenyl)
RN  - EC 3.- (Hydrolases)
SB  - IM
MH  - Arthrobacter/*enzymology/genetics
MH  - Biotransformation
MH  - Biphenyl Compounds/*metabolism
MH  - Catalytic Domain
MH  - Cloning, Molecular
MH  - DNA Mutational Analysis
MH  - Enzyme Inhibitors/analysis
MH  - Enzyme Stability
MH  - Escherichia coli/genetics/metabolism
MH  - Fungicides, Industrial/*metabolism
MH  - Gene Expression
MH  - Hydrogen-Ion Concentration
MH  - Hydrolases/genetics/*metabolism
MH  - Kinetics
MH  - Mutagenesis, Site-Directed
MH  - Temperature
OTO - NOTNLM
OT  - Arthrobacter sp. YC-RL1
OT  - Biphenyl degradation
OT  - BphD
OT  - Enzymatic properties
OT  - Site-direct mutagenesis
EDAT- 2019/06/27 06:00
MHDA- 2019/12/31 06:00
CRDT- 2019/06/27 06:00
PHST- 2019/02/23 00:00 [received]
PHST- 2019/05/28 00:00 [accepted]
PHST- 2019/05/25 00:00 [revised]
PHST- 2019/06/27 06:00 [pubmed]
PHST- 2019/12/31 06:00 [medline]
PHST- 2019/06/27 06:00 [entrez]
AID - 10.1007/s00253-019-09956-z [pii]
AID - 10.1007/s00253-019-09956-z [doi]
PST - ppublish
SO  - Appl Microbiol Biotechnol. 2019 Aug;103(16):6825-6836. doi: 
      10.1007/s00253-019-09956-z. Epub 2019 Jun 26.