ARTP诱变黑曲霉高产β-葡萄糖苷酶的研究

Study on ARTP Mutagenesis of Aspergillus NigerβHigh Yield of Glucosidase

通过ARTP技术操作快速获得突变菌株,提取纤维质中的β-葡萄糖苷酶产菌,进行产酶液体发酵并提取粗酶液,结果可见35~80℃时底物与酶液反应,得酶活性最大数值100%,β-葡萄糖苷酶促反应最为适宜的温度65℃。测定酶活性,最大100%,β-葡萄糖苷酶酶促反应最为适宜pH为4.5。50℃进行酶活性测量,得到100%。实验得出,β-葡萄糖苷酶120min保温之后酶活性可达61.6%。酶液稀释,测量酶活性,最大数值100%。β-葡萄糖苷酶最为稳定状态的ph值为3.0~l0.0,在ph值低于11.0时,β-葡萄糖苷酶活性下降,相对活性90.6%。β-葡萄糖苷酶最为适宜的反应温度为65℃。5mmol/L浓度之下得出不同金属离子,测量酶活性,得100%。Mn2+对酶活性促进作用最强。Cu2+抑制作用最强,此次研究证明β-葡萄糖苷酶pH较为稳定,热稳定性较好,应用价值较大。

A mutant strain was quickly obtained by ARTP techniques,β-glucosidase producing bacteria from the extracts,Fermentation and extraction of crude enzyme solution,The results showed that the substrate reacted with the enzyme solution at 35~80℃,The maximum enzyme activity was 100%,β-most suitable temperature for glucoside enzymatic reaction is 65℃.Determination of enzyme activity,Maximum 100%,β-glucosidase enzymatic reaction is the most suitable PH value 4.5.50℃Enzyme activity was measured,Get 100 percent.The experiment shows,The enzyme activityβ-glucosidase 120 min can reach 61.6%after heat preservation.Enzyme dilution,Measure enzyme activity,The maximum value is 100.The ph valueβ-the most stable state of glucosidase is~l0.0,3.0 At a ph below 11.0,β-glucosidase activity decreased,The relative activity is 90.6.The most suitable reaction temperature forβ-glucosidase is 65℃.5 mmol L-1 Different metal ions are obtained at final concentration,Measure enzyme activity,It’s 100 percent.Mn2 promotes enzyme activity most.Cu2 had the strongest inhibition,This study proved that theβ-glucosidase pH value is relatively stable,Good thermal stability,The application value is great.