当代化工
噹代化工
당대화공
CONTEMPORARY CHEMICAL INDUSTRY
2013年
4期
401-405
,共5页
周洪波%林福来%郑甲%郭宁%吴俊子%吴丽双%邱冠周
週洪波%林福來%鄭甲%郭寧%吳俊子%吳麗雙%邱冠週
주홍파%림복래%정갑%곽저%오준자%오려쌍%구관주
β-甘露聚糖酶%qPCR%mRNA表达%培养温度
β-甘露聚糖酶%qPCR%mRNA錶達%培養溫度
β-감로취당매%qPCR%mRNA표체%배양온도
β-mannanase%QPCR%Pichia pastoris%Culture temperature
β-甘露聚糖酶已经广泛的在毕赤酵母中异源表达,为了提高β-甘露聚糖酶在毕赤酵母中的产量,本文研究了培养温度对β-甘露聚糖酶在毕赤酵母中表达的影响.通过比较不同温度下β-甘露聚糖酶在转录和蛋白表达水平的差异,发现降低温度虽然降低了mRNA表达量,但是外源蛋白得到积累增加;降低培养温度可以显著提高重组菌株的活性,从而减少细胞死亡带来的胞内蛋白酶的释放,最终使得低温下发酵液中的蛋白酶活力显著下降.研究结果表明降低培养温度能显著提高β-甘露聚糖酶在毕赤酵母中的表达量.
β-甘露聚糖酶已經廣汎的在畢赤酵母中異源錶達,為瞭提高β-甘露聚糖酶在畢赤酵母中的產量,本文研究瞭培養溫度對β-甘露聚糖酶在畢赤酵母中錶達的影響.通過比較不同溫度下β-甘露聚糖酶在轉錄和蛋白錶達水平的差異,髮現降低溫度雖然降低瞭mRNA錶達量,但是外源蛋白得到積纍增加;降低培養溫度可以顯著提高重組菌株的活性,從而減少細胞死亡帶來的胞內蛋白酶的釋放,最終使得低溫下髮酵液中的蛋白酶活力顯著下降.研究結果錶明降低培養溫度能顯著提高β-甘露聚糖酶在畢赤酵母中的錶達量.
β-감로취당매이경엄범적재필적효모중이원표체,위료제고β-감로취당매재필적효모중적산량,본문연구료배양온도대β-감로취당매재필적효모중표체적영향.통과비교불동온도하β-감로취당매재전록화단백표체수평적차이,발현강저온도수연강저료mRNA표체량,단시외원단백득도적루증가;강저배양온도가이현저제고중조균주적활성,종이감소세포사망대래적포내단백매적석방,최종사득저온하발효액중적단백매활력현저하강.연구결과표명강저배양온도능현저제고β-감로취당매재필적효모중적표체량.
β-Mannanase has been extensively expressed heterologously in Pichia pastoris. The effect of the culture temperature on the expression ofβ-mannanase in Pichia pastoris was investigated in order to improve the production ofβ-mannanase in Pichia pastoris. Through comparing the difference of transcription and protein expression levels ofβ-mannanase under different culture temperature, it’s found that mRNA level decreased with decreasing of the temperature, but exogenous protein was accumulated. Lowering culture temperature could evidently improve the cell aviability of the recombinant strain, which could reduce the release of intracellular protease from the death cell, ultimately make the protease activity in the fermentation broth decrease significantly under the low temperature cultivate condition. The research results show that lowering culture temperature can evidently improve the expression ofβ-mannanase in Pichia pastoris.