The recombinant expression, purification and activity analysis of lactate dehydrogenase in Veillonella dispar

VernacularTitle:殊异韦荣菌中乳酸脱氢酶重组蛋白的表达、纯化及活性分析
Author: Zhong-Qin HE ¹ ; Cheng ZHONG ; Xin GAO ; Ying XUE ; Xiao-Yu SUN ; Xiao-Hong LIU
Author Information

1. 吉林大学中日联谊医院口腔科
Keywords: Veillonella; Lactate dehydrogenases; Preliminary expression; Protein activity
From: Chinese Journal of Stomatology 2012;47(z1):63-67
CountryChina
Language:Chinese
Abstract: Objective To determine dependence lactate dehydrogenase activity in Veillonella dispar.Methods Transformed the recombinant plasmid pET-28a-LDH in BL21 (DE3) and Rosetta(DE3).Recombinant lactate dehydrogenase(LDH) protein was induced at different conditions.Induction condition was optimized to obtain proper yield of recombinant protein.After purification by HisTRAP FF column,the protein activity was determined with LDH reactive kit.Results Sodium dodecylsulfate-polyacrylamide gel electrophoresis showed that the best protein expression conditions were isopropylthio-β-D-galactoside (IPTG)end for 0.4 mmol/L concentration in 30 degrees to induce 8 hours,or IPTG end for 0.4 mmol/L concentration in 30 degrees to induce 6 hours.Through the analysis of BandScan 5.0,the expression in BL21 (DE3) was 9.0％,higher than 6.1％ in Rosetta(DE3).After purification by HisTRAP FF column,the protein active value of 13.79.Conclusions Recombinant LDH protein could be induced by IPTG with an optimal condition.