| dc.creator | Li,Wei | |
| dc.creator | Ng,I-Son | |
| dc.creator | Fang,Baishan | |
| dc.creator | Yu,Jincong | |
| dc.creator | Zhang,Guangya | |
| dc.date | 2011-07-01 | |
| dc.date.accessioned | 2019-05-03T12:44:52Z | |
| dc.date.available | 2019-05-03T12:44:52Z | |
| dc.identifier | https://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582011000400007 | |
| dc.identifier.uri | http://revistaschilenas.uchile.cl/handle/2250/85221 | |
| dc.description | The gene dhaT from Klebsiella pneumoniae encoding 1,3-propanediol oxidoreductase (PDOR) was de novo synthesized by splicing overlap extension polymerase chain reaction (SOE-PCR) primarily according to Escherichia coli s codon usage, as well as mRNA secondary structure. After optimization, Codon Adaptation Index (CAI) value was improved from 0.75 to 0.83, meanwhile energy of mRNA secondary structure was increased from -400.1 to -86.8 kcal/mol. This synthetic DNA was under control by phage T7 promoter in the expression vector pET-15b and transformed into the E. coli BL21 (DE3) strain. Inducers such as isopropyl β-D-thiogalactoside (IPTG) and lactose were compared by activity at different inducing time. The activity of PDOR after codon optimized was 385.4 ± 3.6 U/mL, which was almost 5-fold higher than wild type (82.3 ± 1.5 U/ml) under the flask culture at 25ºC for 10 hrs. Then his-tagged enzyme was separated by using Ni-IDA column. The favorite environment for enzyme activity was at 5°C and pH 10.0, PDOR showed a certainly stability in potassium carbonate buffer for 2 hrs at diverse temperatures, enzyme activity was significantly improved by Mn2+. | |
| dc.format | text/html | |
| dc.language | en | |
| dc.publisher | Pontificia Universidad Católica de Valparaíso | |
| dc.rights | info:eu-repo/semantics/openAccess | |
| dc.source | Electronic Journal of Biotechnology v.14 n.4 2011 | |
| dc.subject | 1,3-propanediol oxidoreductase | |
| dc.subject | codon optimization | |
| dc.subject | enzymatic properties | |
| dc.subject | Escherichia coli | |
| dc.subject | overlap extension PCR | |
| dc.title | Codon optimization of 1,3-propanediol oxidoreductase expression in Escherichia coli and enzymatic properties | |
