安徽农学通报
安徽農學通報
안휘농학통보
AUHUI AGRICULTURAL SCIENCE BULLETIN
2012年
21期
63-66
,共4页
陈龙军%陈亚兰%陈济琛%林新坚
陳龍軍%陳亞蘭%陳濟琛%林新堅
진룡군%진아란%진제침%림신견
毕赤酵母β—1%3-1%4-葡聚糖酶%发酵%酶学性质
畢赤酵母β—1%3-1%4-葡聚糖酶%髮酵%酶學性質
필적효모β—1%3-1%4-포취당매%발효%매학성질
Pichia pastoris%β - 1,3 - 1,4 - glucanase%Fermentation%Characterization
对毕赤酵母表达β-1,3—1,4-葡聚糖酶条件进行优化,在摇瓶水平上研究了温度、pH、甲醇流加量、诱导时间,油酸等因素对重组葡聚糖酶的影响;得优化条件为:最适温度30℃、pH6.0,最佳甲醇诱导浓度为0.5%,最佳诱导时间为84h,酶活力达27.4U/mL,比初始酶活力提高了1.9倍。酶反应的最适pH为6.0,最适温度为50℃。在pH3.5—8.0的范围内和温度55℃以下保存时具有较好的稳定性;其中,CaCl2对重组酶的激活效果显著,使酶活力提高达92%。
對畢赤酵母錶達β-1,3—1,4-葡聚糖酶條件進行優化,在搖瓶水平上研究瞭溫度、pH、甲醇流加量、誘導時間,油痠等因素對重組葡聚糖酶的影響;得優化條件為:最適溫度30℃、pH6.0,最佳甲醇誘導濃度為0.5%,最佳誘導時間為84h,酶活力達27.4U/mL,比初始酶活力提高瞭1.9倍。酶反應的最適pH為6.0,最適溫度為50℃。在pH3.5—8.0的範圍內和溫度55℃以下保存時具有較好的穩定性;其中,CaCl2對重組酶的激活效果顯著,使酶活力提高達92%。
대필적효모표체β-1,3—1,4-포취당매조건진행우화,재요병수평상연구료온도、pH、갑순류가량、유도시간,유산등인소대중조포취당매적영향;득우화조건위:최괄온도30℃、pH6.0,최가갑순유도농도위0.5%,최가유도시간위84h,매활력체27.4U/mL,비초시매활력제고료1.9배。매반응적최괄pH위6.0,최괄온도위50℃。재pH3.5—8.0적범위내화온도55℃이하보존시구유교호적은정성;기중,CaCl2대중조매적격활효과현저,사매활력제고체92%。
In this word, the express conditions ofβ - 1, 3 - 1, 4 - glucanase are optimized in Pichia pastoris. By the shakeing flask, the infuluences of temperature, pH, methanol, inducing time, oleic acid on the expression of glucanase were investigated. The results show that the optimal cultural conditions were temperature 30℃, pH6.0, and best methanol con- centration was 0.5%, induction time for 84 h, enzyme activity reached 27.4 U/mL. The optimal reaction pH and tempera- ture of glucanase were 6.0 and 50℃ respectively. The enzyme was stable within pH3.5 - 8.0 under 60℃. Furthermore, CaC12 can significantly improve the enzyme activity, make the enzyme activity increase 92%.