schliessen

Filtern

 

Bibliotheken

Disorder prediction-based construct optimization improves activity and catalytic efficiency of Bacillus naganoensis pullulanase

Pullulanase is a well-known starch-debranching enzyme. However, the production level of pullulanase is yet low in both wide-type strains and heterologous expression systems. We predicted the disorder propensities of Bacillus naganoensis pullulanase (PUL) using the bioinformatics tool, Disorder Predi... Full description

Journal Title: Sci Rep 2016, Vol.6(1), pp.24574-24574
Main Author: Wang, Xinye
Other Authors: Nie, Yao , Mu, Xiaoqing , Xu, Yan , Xiao, Rong
Format: Electronic Article Electronic Article
Language: English
Subjects:
ID: ISSN: 2045-2322 ; DOI: 10.1038/srep24574
Zum Text:
SendSend as email Add to Book BagAdd to Book Bag
Staff View
recordid: palgrave_j10.1038/srep24574
title: Disorder prediction-based construct optimization improves activity and catalytic efficiency of Bacillus naganoensis pullulanase
format: Article
creator:
  • Wang, Xinye
  • Nie, Yao
  • Mu, Xiaoqing
  • Xu, Yan
  • Xiao, Rong
subjects:
  • Article
ispartof: Sci Rep, 2016, Vol.6(1), pp.24574-24574
description: Pullulanase is a well-known starch-debranching enzyme. However, the production level of pullulanase is yet low in both wide-type strains and heterologous expression systems. We predicted the disorder propensities of Bacillus naganoensis pullulanase (PUL) using the bioinformatics tool, Disorder Prediction Meta-Server. On the basis of disorder prediction, eight constructs, including PULΔN5, PULΔN22, PULΔN45, PULΔN64, PULΔN78 and PULΔN106 by deleting the first 5, 22, 45, 64, 78 and 106 residues from the N-terminus and PULΔC9 and PULΔC36 by deleting the last 9 and 36 residues from the C-terminus, were cloned into the recombinant expression vector pET-28a-PelB and auto-induced in Escherichia coli BL21 (DE3) cells. All constructs were evaluated in production level, specific activities and kinetic parameters. Both PULΔN5 and PULΔN106 gave higher production levels of protein than the wide type and displayed increased specific activities. Kinetic studies showed that substrate affinities of the mutants were improved in various degrees and the catalytic efficiency of PULΔN5, PULΔN45, PULΔN78, PULΔN106 and PULΔC9 were enhanced. However, the truncated mutations did not change the advantageous properties of the enzyme involving optimum temperature and pH for further application. Therefore, Disorder prediction-based truncation would be helpful to efficiently improve the enzyme activity and catalytic efficiency.
language: eng
source:
identifier: ISSN: 2045-2322 ; DOI: 10.1038/srep24574
fulltext: fulltext
issn:
  • 2045-2322
  • 20452322
url: Link


@attributes
ID398861294
RANK0.07
NO1
SEARCH_ENGINEprimo_central_multiple_fe
SEARCH_ENGINE_TYPEPrimo Central Search Engine
LOCALfalse
PrimoNMBib
record
control
sourcerecordid10.1038/srep24574
sourceidpalgrave_j
recordidTN_palgrave_j10.1038/srep24574
sourcesystemOther
pqid1782831075
display
typearticle
titleDisorder prediction-based construct optimization improves activity and catalytic efficiency of Bacillus naganoensis pullulanase
creatorWang, Xinye ; Nie, Yao ; Mu, Xiaoqing ; Xu, Yan ; Xiao, Rong
ispartofSci Rep, 2016, Vol.6(1), pp.24574-24574
identifierISSN: 2045-2322 ; DOI: 10.1038/srep24574
descriptionPullulanase is a well-known starch-debranching enzyme. However, the production level of pullulanase is yet low in both wide-type strains and heterologous expression systems. We predicted the disorder propensities of Bacillus naganoensis pullulanase (PUL) using the bioinformatics tool, Disorder Prediction Meta-Server. On the basis of disorder prediction, eight constructs, including PULΔN5, PULΔN22, PULΔN45, PULΔN64, PULΔN78 and PULΔN106 by deleting the first 5, 22, 45, 64, 78 and 106 residues from the N-terminus and PULΔC9 and PULΔC36 by deleting the last 9 and 36 residues from the C-terminus, were cloned into the recombinant expression vector pET-28a-PelB and auto-induced in Escherichia coli BL21 (DE3) cells. All constructs were evaluated in production level, specific activities and kinetic parameters. Both PULΔN5 and PULΔN106 gave higher production levels of protein than the wide type and displayed increased specific activities. Kinetic studies showed that substrate affinities of the mutants were improved in various degrees and the catalytic efficiency of PULΔN5, PULΔN45, PULΔN78, PULΔN106 and PULΔC9 were enhanced. However, the truncated mutations did not change the advantageous properties of the enzyme involving optimum temperature and pH for further application. Therefore, Disorder prediction-based truncation would be helpful to efficiently improve the enzyme activity and catalytic efficiency.
languageeng
source
subjectArticle;
version7
lds50peer_reviewed
links
openurl$$Topenurl_article
openurlfulltext$$Topenurlfull_article
search
creatorcontrib
0Wang, Xinye
1Nie, Yao
2Mu, Xiaoqing
3Xu, Yan
4Xiao, Rong
titleDisorder prediction-based construct optimization improves activity and catalytic efficiency of Bacillus naganoensis pullulanase
descriptionPullulanase is a well-known starch-debranching enzyme. However, the production level of pullulanase is yet low in both wide-type strains and heterologous expression systems. We predicted the disorder propensities of Bacillus naganoensis pullulanase (PUL) using the bioinformatics tool, Disorder Prediction Meta-Server. On the basis of disorder prediction, eight constructs, including PULΔN5, PULΔN22, PULΔN45, PULΔN64, PULΔN78 and PULΔN106 by deleting the first 5, 22, 45, 64, 78 and 106 residues from the N-terminus and PULΔC9 and PULΔC36 by deleting the last 9 and 36 residues from the C-terminus, were cloned into the recombinant expression vector pET-28a-PelB and auto-induced in Escherichia coli BL21 (DE3) cells. All constructs were evaluated in production level, specific activities and kinetic parameters. Both PULΔN5 and PULΔN106 gave higher production levels of protein than the wide type and displayed increased specific activities. Kinetic studies showed that substrate affinities of the mutants were improved in various degrees and the catalytic efficiency of PULΔN5, PULΔN45, PULΔN78, PULΔN106 and PULΔC9 were enhanced. However, the truncated mutations did not change the advantageous properties of the enzyme involving optimum temperature and pH for further application. Therefore, Disorder prediction-based truncation would be helpful to efficiently improve the enzyme activity and catalytic efficiency.
general
0English
110.1038/srep24574
2Palgrave Macmillan Journals
sourceidpalgrave_j
recordidpalgrave_j10.1038/srep24574
issn
02045-2322
120452322
rsrctypearticle
creationdate2016
addtitleSci Rep
searchscopepalgrave_j
scopepalgrave_j
lsr30VSR-Enriched:[pages, subject, date, pqid]
sort
titleDisorder prediction-based construct optimization improves activity and catalytic efficiency of Bacillus naganoensis pullulanase
authorWang, Xinye ; Nie, Yao ; Mu, Xiaoqing ; Xu, Yan ; Xiao, Rong
facets
frbrgroupid-2254154829878927108
frbrtype5
newrecords20180122
languageeng
creationdate2016
collectionPalgrave Macmillan Journals
prefilterarticles
rsrctypearticles
creatorcontrib
0Wang, Xinye
1Nie, Yao
2Mu, Xiaoqing
3Xu, Yan
4Xiao, Rong
jtitleSci Rep
toplevelpeer_reviewed
delivery
delcategoryRemote Search Resource
fulltextfulltext
addata
aulast
0Wang
1Nie
2Mu
3Xu
4Xiao
aufirst
0Xinye
1Yao
2Xiaoqing
3Yan
4Rong
au
0Wang, Xinye
1Nie, Yao
2Mu, Xiaoqing
3Xu, Yan
4Xiao, Rong
atitleDisorder prediction-based construct optimization improves activity and catalytic efficiency of Bacillus naganoensis pullulanase
jtitleSci Rep
risdate20169
volume6
issue1
spage24574
epage24574
issn2045-2322
formatjournal
genrearticle
ristypeJOUR
abstractPullulanase is a well-known starch-debranching enzyme. However, the production level of pullulanase is yet low in both wide-type strains and heterologous expression systems. We predicted the disorder propensities of Bacillus naganoensis pullulanase (PUL) using the bioinformatics tool, Disorder Prediction Meta-Server. On the basis of disorder prediction, eight constructs, including PULΔN5, PULΔN22, PULΔN45, PULΔN64, PULΔN78 and PULΔN106 by deleting the first 5, 22, 45, 64, 78 and 106 residues from the N-terminus and PULΔC9 and PULΔC36 by deleting the last 9 and 36 residues from the C-terminus, were cloned into the recombinant expression vector pET-28a-PelB and auto-induced in Escherichia coli BL21 (DE3) cells. All constructs were evaluated in production level, specific activities and kinetic parameters. Both PULΔN5 and PULΔN106 gave higher production levels of protein than the wide type and displayed increased specific activities. Kinetic studies showed that substrate affinities of the mutants were improved in various degrees and the catalytic efficiency of PULΔN5, PULΔN45, PULΔN78, PULΔN106 and PULΔC9 were enhanced. However, the truncated mutations did not change the advantageous properties of the enzyme involving optimum temperature and pH for further application. Therefore, Disorder prediction-based truncation would be helpful to efficiently improve the enzyme activity and catalytic efficiency.
pubNature Publishing Group UK
doi10.1038/srep24574
pages24574
date2016-04-19