PMID- 27479442
OWN - NLM
STAT- MEDLINE
DCOM- 20170822
LR  - 20210405
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 11
IP  - 8
DP  - 2016
TI  - Surface Immobilization of Human Arginase-1 with an Engineered Ice Nucleation 
      Protein Display System in E. coli.
PG  - e0160367
LID - 10.1371/journal.pone.0160367 [doi]
LID - e0160367
AB  - Ice nucleation protein (INP) is frequently used as a surface anchor for protein 
      display in gram-negative bacteria. Here, MalE and TorA signal peptides, and three 
      charged polypeptides, 6xLys, 6xGlu and 6xAsp, were anchored to the N-terminus of 
      truncated INP (InaK-N) to improve its surface display efficiency for human 
      Arginase1 (ARG1). Our results indicated that the TorA signal peptide increased 
      the surface translocation of non-protein fused InaK-N and human ARG1 fused InaK-N 
      (InaK-N/ARG1) by 80.7% and 122.4%, respectively. Comparably, the MalE signal 
      peptide decreased the display efficiencies of both the non-protein fused InaK-N 
      and InaK-N/ARG1. Our results also suggested that the 6xLys polypeptide 
      significantly increased the surface display efficiency of K6-InaK-N/ARG1 by 
      almost 2-fold, while also practically abolishing the surface translocation of 
      non-protein fused InaK-N, indicating the interesting roles of charged 
      polypeptides in bacteria surface display systems. Cell surface-immobilized 
      K6-InaK-N/ARG1 presented an arginase activity of 10.7 U/OD600 under the optimized 
      conditions of 40 degrees C, pH 10.0 and 1 mM Mn2+, which could convert more than 95% of 
      L-Arginine (L-Arg) to L-Ornithine (L-Orn) in 16 hours. The engineered InaK-Ns 
      expanded the INP surface display system, which aided in the surface 
      immobilization of human ARG1 in E. coli cells.
FAU - Zhang, Zhen
AU  - Zhang Z
AD  - Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, 
      Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei 
      University, Wuhan, People's Republic of China.
FAU - Tang, Rongxin
AU  - Tang R
AD  - Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, 
      Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei 
      University, Wuhan, People's Republic of China.
FAU - Bian, Lu
AU  - Bian L
AD  - Beihai Blood Center, Guang Xi, People's Republic of China.
FAU - Mei, Meng
AU  - Mei M
AD  - Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, 
      Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei 
      University, Wuhan, People's Republic of China.
FAU - Li, Chunhua
AU  - Li C
AD  - Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, 
      Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei 
      University, Wuhan, People's Republic of China.
FAU - Ma, Xiangdong
AU  - Ma X
AD  - Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, 
      Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei 
      University, Wuhan, People's Republic of China.
FAU - Yi, Li
AU  - Yi L
AD  - Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, 
      Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei 
      University, Wuhan, People's Republic of China.
FAU - Ma, Lixin
AU  - Ma L
AD  - Hubei Collaborative Innovation Center for Green Transformation of Bio-resources, 
      Hubei Key Laboratory of Industrial Biotechnology, College of Life Sciences, Hubei 
      University, Wuhan, People's Republic of China.
LA  - eng
PT  - Journal Article
DEP - 20160801
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Bacterial Outer Membrane Proteins)
RN  - 0 (Bacterial Proteins)
RN  - 0 (Oligopeptides)
RN  - 0 (Protein Sorting Signals)
RN  - 0 (ice nucleation protein)
RN  - E524N2IXA3 (Ornithine)
RN  - EC 3.5.3.1 (ARG1 protein, human)
RN  - EC 3.5.3.1 (Arginase)
RN  - K3Z4F929H6 (Lysine)
SB  - IM
MH  - Arginase/genetics/*metabolism
MH  - Bacterial Outer Membrane Proteins/genetics/*metabolism
MH  - Bacterial Proteins/genetics/*metabolism
MH  - Escherichia coli/*metabolism
MH  - Flow Cytometry
MH  - Genetic Vectors/genetics/metabolism
MH  - Humans
MH  - Lysine/genetics/metabolism
MH  - Microscopy, Fluorescence
MH  - Oligopeptides/genetics/metabolism
MH  - Ornithine/metabolism
MH  - Protein Engineering
MH  - Protein Sorting Signals/genetics
MH  - Translocation, Genetic
PMC - PMC4968799
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2016/08/02 06:00
MHDA- 2017/08/23 06:00
PMCR- 2016/08/01
CRDT- 2016/08/02 06:00
PHST- 2016/02/22 00:00 [received]
PHST- 2016/07/18 00:00 [accepted]
PHST- 2016/08/02 06:00 [entrez]
PHST- 2016/08/02 06:00 [pubmed]
PHST- 2017/08/23 06:00 [medline]
PHST- 2016/08/01 00:00 [pmc-release]
AID - PONE-D-16-07541 [pii]
AID - 10.1371/journal.pone.0160367 [doi]
PST - epublish
SO  - PLoS One. 2016 Aug 1;11(8):e0160367. doi: 10.1371/journal.pone.0160367. 
      eCollection 2016.