黑曲霉木聚糖酶的同源表达、发酵及其酶学性质研究-动物遗传育种与繁殖专业论文 word格式.docx

优秀毕业论文 精品参考文献资料 A327. About more than one hundred transformants was obtained. 96 strains of them were screened by shake flask, the results show that 87.5% transformants have higher enzyme activity comparing with the original strain A327. By PCR technology, a 1.8kb fragment in the selectable marker genes amds was amplified from chromosome DNA of part transformations, this result indicated that the expression was positive expression. We Selected 8 transformants that have higher xylanase activity than others to isolate single-spore on plate and to test enzyme productivities by liquid fermentation, finding that xylanase activity of NO.70 was the highest. The fermentation products of part transformants and original strain A327 was analyzed with SDSelectrophoresis, the result is accordant with the outcome of assaying xylanase activity by liquid fermentation. Study on liquid fermentation of bioengineered strain 70# producing xylanase On the basis of a single factor experiment, the optimum culture medium of 70# strain was investigated by the orthogonal experiment in four factors and three levels and the optimal conditions of cultivation was studied. We ascertained the optimal culture medium and fermentation condition for producing xylanase by transformant 70#. The main experimental results were summarized as following: The optimum culture medium and optimal cultivation condition are(g/L): corn core 80, wheat bran 18, gooey 14, NaNO3 8. It cultivated in shaker at 28～31℃ for 72h，5% spores were inoculated into 30ml medium in 100mL Erlenmeyer flask and the rotate speed is 230rpm. Under these conditions, the xylanase activity of 70# is as 30 times as that of the original strain A327 and the maximal xylanase activity was 6102IU/mL. Purification and characterization of xylanase produced by transformant 70# A component of xylanase was separated by ammonium sulfate precipitation, followed by dialysis overnight and then through DEAE Sepharose anion exchange chromatography from the l