高温淀粉酶高产菌株筛选及产酶条件优化.docVIP

高温淀粉酶高产菌株筛选及产酶条件优化 　　摘要 为筛选高温淀粉酶高产菌株，利用淀粉酶水解圈作筛选模型，从稻田中采集土样，筛选得到产淀粉酶能力较高的菌株E10。为提高菌株产酶能力，进行了碳源种类及浓度，氮源种类及浓度，无机盐种类及浓度，pH值，种龄及接种量、温度、时间的单因素试验，并对发酵培养基及发酵条件进行了L9（34）正交优化试验。结果表明：最佳产酶发酵培养基为2.0%麸皮、0.1%硝酸钾、0.25%氯化钙，最适初始pH值为6.0。最适产酶条件为种龄24 h，接种量7 mL，于36 ℃下培养48 h。在优化条件下，菌株产酶活力可达180.94 IU/mL。 　　关键词 高温淀粉酶；高产菌株；筛选；发酵条件 　　中图分类号 TQ925+.1 文献标识码 A 文章编号 1007-5739（2014）07-0240-04 　　Screening of High-yield Strain Producing Thermostable Amylase and Optimization of Enzyme Production 　　YI Zheng-xuan ZHAO Tong SHI Xuan WANG Zheng LI Li-heng * 　　（College of Bioscience and Technology，Hunan Agricultural University，Changsha Hunan 410128） 　　Abstract In order to screen a high-yield strain producing thermostable amylase，starch hydrolysis circle was employed as a screening model. The E10 strain achieved from the paddy field soil has high capacity of enzyme production. In order to improve the capability of enzyme production of E10，the single factor experiments was conducted for the kinds and concentrations of carbon source，the kinds and concentrations of nitrogen source，the kinds and concentrations of inorganic salt，pH value，seed age，inoculate volume，temperature and time，then the L9（34） orthogonal optimization test for the fermentation medium and conditions was carried out. The results showed that the best fermentation medium of the enzyme production of E10 was 2.0% wheat bran，0.1% KNO3，0.25% CaCl2，initial pH value of 6.0. The optimal fermentation conditions of the enzyme production of E10 were 24 hours of seed age，7 mL of inoculates volume，36 ℃ of culture temperature，48 hours of time. Under the optimized conditions，the enzyme production of E10 reached 180.94 IU/mL. 　　Key words thermostable amylase；high-yield strain；screening；fermentation condition 　　淀粉酶是水解淀粉和糖原的酶类总称，广泛分布于动物（唾液、胰脏等）、植物（麦芽、山?菜）及微生物中，是最早实现工业生产并且迄今为止用途最广、产量最大的酶制剂品种[1]。20世纪60年代以来，由于酶法生产葡萄糖，以及用葡萄糖生产异构糖浆的大规模工业化生产，淀粉酶的需求量越来越大，几乎占酶制剂总产量的50%以上。淀粉酶为作用于淀粉、糖原等α-1，4-葡聚糖，水解α-1，4-糖苷键的酶。根据酶水解产物异构类型的不同可分为α-淀粉酶（EC3.2.1.1）与β-淀粉酶（EC3.2.1.