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dc.creatorBai,Xi
dc.creatorYuan,Xianjun
dc.creatorWen,Aiyou
dc.creatorLi,Junfeng
dc.creatorBai,Yunfeng
dc.creatorShao,Tao
dc.date2016-11-01
dc.date.accessioned2019-05-03T12:45:28Z
dc.date.available2019-05-03T12:45:28Z
dc.identifierhttps://scielo.conicyt.cl/scielo.php?script=sci_arttext&pid=S0717-34582016000600012
dc.identifier.urihttp://revistaschilenas.uchile.cl/handle/2250/85586
dc.descriptionBackground: Cold-active endo-1, 4-β-glucanase (EglC) can decrease energy costs and prevent product denaturation in biotechnological processes. However, the nature EglC from C. farmeri A1 showed very low activity (800 U/L). In an attempt to increase its expression level, C. farmeri EglC was expressed in Escherichia coli as an N-terminal fusion to protein S (ProS) from Myxococcus xanthus. Results: A novel expression vector, pET(ProS-EglC), was successfully constructed for the expression of C. farmeri EglC in E. coli. SDS-PAGE showed that the recombinant protein (ProS-EglC) was approximately 60 kDa. The activity of ProS-EglC was 12,400 U/L, which was considerably higher than that of the nature EglC (800 U/L). ProS-EglC was active at pH 6.5-pH 8.0, with optimum activity at pH 7.0. The recombinant protein was stable at pH 3.5-pH 6.5 for 30 min. The optimal temperature for activity of ProS-EglC was 30°C-40°C. It showed greater than 50% of maximum activity even at 5°C, indicating that the ProS-EglC is a cold-active enzyme. Its activity was increased by Co2+ and Fe2+, but decreased by Cd2+, Zn2+, Li+, methanol, Triton-X-100, acetonitrile, Tween 80, and SDS. Conclusions: The ProS-EglC is promising in application of various biotechnological processes because of its cold-active characterizations. This study also suggests a useful strategy for the expression of foreign proteins in E. coli using a ProS tag.
dc.formattext/html
dc.languageen
dc.publisherPontificia Universidad Católica de Valparaíso
dc.relation10.1016/j.ejbt.2016.10.005
dc.rightsinfo:eu-repo/semantics/openAccess
dc.sourceElectronic Journal of Biotechnology v.19 n.6 2016
dc.subjectCellulose degradation
dc.subjectCellulose
dc.subjectCold-active enzyme
dc.subjectEndoglucanases
dc.subjectEnzymatic properties
dc.subjectEscherichia coli
dc.subjectExpression
dc.subjectNovel expression vector
dc.subjectN-terminal fusion
dc.subjectProtein S-tag
dc.subjectRecombinant protein
dc.titleEfficient expression and characterization of a cold-active endo-1, 4-β-glucanase from Citrobacter farmeri by co-expression of Myxococcus xanthus protein S


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