中文摘要：

从Bacillus subtilis JNA 3-10中克隆出β-甘露聚糖酶基因成熟肽链编码序列manA1和含信号肽的β-甘露聚糖酶基因manA2，在B. subtilis 168中克隆表达，分别筛选获得高效分泌表达β-甘露聚糖酶的重组菌株BPM1001（pMA5-manA1/ B. subtilis 168）和BPM1002（pMA5-manA2/B. subtilis 168），结果表明菌株BPM1002总酶活力是菌株BPM1001的9.65倍，是原始菌株的13.1倍. 在基因manA2下游引入His序列克隆出β-甘露聚糖酶基因manA3，获得枯草芽孢杆菌168重组菌株BPM1003. 采用Ni-NTA柱纯化重组菌株BPM1003分泌表达的β-甘露聚糖酶，并研究其酶学性质，该酶促反应的最适pH为6.5，最适温度为65 ℃，在37 ℃条件下保存一个月酶活力依然保留有77.8%. 5 L发酵罐放大实验结果表明魔芋粉对于产β-甘露聚糖酶具有明显的诱导作用，酶活力最高可达2 748.82 U/mL. 图9 表3 参19

英文摘要：

The manA1 gene encoding mature β-mannanase and the manA2 gene contained a signal peptide from the Bacillus subtilis JNA 3-10 were amplified. The two genes were inserted into expression vector pMA5, and the plasmid pMA5- manA1 and pMA5-manA2 were constructed and transformed into B. subtilis 168 respectively. The recombinant strains BPM1001 （pMA5-manA1 / B. subtilis 168） and BPM1002 （pMA5-manA2/B. subtilis 168） were therefore obtained. SDS-PAGE showed that the genes manA1 and manA2 were expressed successfully in recombinant B. subtilis 168. After incubated in shake-flask, the enzyme activity of strain BPM1002 was 9.65-fold higher than that of BPM1001. Subsequently, the β-mannanase gene manA3 was cloned with His-Tag added downstream of the gene manA2. Then the enzyme was purified successively by Ni affinity chromatography. The analysis of enzymatic properties showed that the optimum activity of the β-mannanase was at pH 6.5 and 65 ℃, and the enzyme activity was maintained 77.8% of original activity after incubated at 37 ℃ for 30 days. The activity of the β-mannanase reached 2 748.82 U/mL in a 5 L fermentor. Fig 9, Tab 3, Ref 19