海洋酵母普鲁兰类酵母一种新型生淀粉酶生产和特性研究-海洋生物学专业论文.docxVIP

原料生产高浓度的粗酶液。通过对固体发酵参数的优化，成功地将发酵提取液中 原料生产高浓度的粗酶液。通过对固体发酵参数的优化，成功地将发酵提取液中 的粗酶比活力提高到了120．5U／mg蛋白。相对于液体发酵粗酶液，酶比活力提高 了一倍。 本实验研究获得了一株高产新型生淀粉酶的海洋酵母菌株N13d，这是首次 关于海洋普鲁兰类酵母产生淀粉酶的报道，开发了新的淀粉酶资源。经过液体发 酵，我们得到了生淀粉酶的粗酶液，粗酶液能够降解高浓度的生土豆淀粉，水解 产物基本全是葡萄糖。纯酶性质的实验结果表明海洋酵母普鲁兰类酵母N13d产 的生淀粉酶与其他酵母和真菌所产的生淀粉酶有很多不同之处，最有特色的有两 点，一是该酶具有很强的降解a．1，6糖苷键的能力；二是该酶对高浓度的生土 豆淀粉同样具有极强的降解能力。N13d产生的生淀粉酶可以在食品和发酵工业 用来水解生土豆淀粉，产物可以直接作为发酵原料，应用前景广阔。本研究表明 海洋酵母具有独特的生理特征，能够产生一些与陆地酵母和真菌所产的酶不同的 新型酶类，为工业生产提供资源。 关键词：海洋酵母，淀粉酶，蛋白纯化，生淀粉，固体发酵 The The study on characteristic and production of a novel raw starch amylase from marine yeast Aureobasidium pullulans Abstract Marine yeasts are a kind of mieobial community living in marine environments which plays a important role in marine eeo-system．]M_arine yeast strains Call produce many enzymes such as protease，amylase,lipase，phytase，inulinase，cellulase， killer toxin and so on．It is an interesting work for l坞to investigate the application of these bioresourees． In this study,marine yeast strain N13d,producing a high amount ofextracellular amylase was isolated from the deep sea sediments ofPacific Ocean．This strain Was identified to be Aureobasidiumpullulans by 18S rRNA gene sequence analysis and routine yeast identification methods．This is the first report that marine Aureobasidium pullulans produced extracellular amylase．Based On optin枷medium and flask cultivation conditions for amylase production by the marine yeast N13d we carried out amylase liquid fermentation experiment by 2L fermentor．Under the optimal aeration rate 6 Lmin一1 and agitation speed 250 rpm the amylase specific activity in a眦enzyme preparation reach 58．5U／mg protein．Hydrolysis ofraw potato starch Was carriedoutat60℃．After6hofincubation,59．5％ofthe rawpotato starchwith 36％concentration Was hydrolyzed．The end product was identified to glucose by TLC andHPLc． In order to explore the potential application ofthis amylase we purified this eozyme and carried out a comprehensive study about the properties ofenzyme．Th