CAJ | 학술논문

从实验室分离保存的1株产果胶酶的菌株(BTC105)中克隆果胶裂解酶基因(PelC)完整开放阅读框,通过载体构建,将目的基因连接到表达载体pET28a上,转化大肠埃希菌BL21(DE3)进行融合表达,在LB(Luria-Bertani)中进行摇瓶发酵,1 mmol/L IPTG(异丙基-β-D-硫代半乳糖苷)诱导.结果表明,构建了表达载体pET28a-pelC,果胶裂解酶主要在胞内表达,酶活最适pH为5.4,最适温度为50℃,Ca~(2+)对酶活促进作用最为明显,Cu~(2+)完全抑制了酶的活性.
종실험실분리보존적1주산과효매적균주(BTC105)중극륭과효렬해매기인(PelC)완정개방열독광,통과재체구건,장목적기인련접도표체재체pET28a상,전화대장애희균BL21(DE3)진행융합표체,재LB(Luria-Bertani)중진행요병발효,1 mmol/L IPTG(이병기-β-D-류대반유당감)유도.결과표명,구건료표체재체pET28a-pelC,과효렬해매주요재포내표체,매활최괄pH위5.4,최괄온도위50℃,Ca~(2+)대매활촉진작용최위명현,Cu~(2+)완전억제료매적활성.
A complete open reading frame of pectin lyase C (PelC) cloned from a pectinase-producing strain BTC105 isolated and collected in the laboratory was constructed on a plasmid pET28a and transferred into E. coli BL21 (DE3) to carry out fuse expression; and flask shaking fermented in LB (Luria-Bertani) , induced with 1 mmol/L IPTG (iso-propyl β-D-1-thiogalactopyranoside). The results showed that the recombinant plasmid pET28a-pelC was constructed successfully and PelC has mainly expressed in E. coli BL21 (DE3). The optimal pH of the enzyme was 5.4, optimal temperature at 50℃, Ca~(2+) stimulated strongly on the enzyme activity, however, Cu~(2+) completely inhibited the activity.