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首页> 外文期刊>African Journal of Biotechnology >Combined strategies for the improvement of heterologous expression of a His-tagged Yarrowia lipolytica lipase Lip2 in Pichia pastoris
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Combined strategies for the improvement of heterologous expression of a His-tagged Yarrowia lipolytica lipase Lip2 in Pichia pastoris

机译：改善巴斯德毕赤酵母中带有组氨酸标签的解脂耶氏酵母脂肪酶Lip2异源表达的联合策略

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Yarrowia lipolytica?lipase Lip2 (YlLip2) is an important biocatalyst for ester synthesis,biodiesel production and enantiomer resolution.?The YlLip2 with an N-terminal histidine-tag?(His6-YlLip2)?was successfully expressed in?Pichia pastoris. Threedifferent cultivation?strategies?had been compared for the production of?His6-YlLip2?byP. pastoris?using a 10-l bioreactor.?The results showed that?His6-YlLip2?activity and cell?viability?could be greatly improved by employing the combined strategies.?Using a low salt medium?(LSM)?instead of the?basal salt medium (BSM)?and?lowering the temperature from 28 to 25°C,?the maximum His6-YlLip2 activity and volumetric productivity were respectively increased by 55.3 and 79.8%. The production of His6-YlLip2 and cell viability was further improved by combining sorbitol co-feeding with methanol. In this culture strategy, the maximum activity of His6-YlLip2?reached15,600?U/ml?after 114 h of induction. The cell mortality?decreased by?11.2% (while thecontrol decreased about?27.6%) after 120 h methanol induction.?The N-terminal histidine-tag brought convenience to purification. The molecular weight of His6-YlLip2 was about 38 kDa. The pure His6-YlLip2 presented a specific activity of 4,830 U/mg when olive oil was used as the substrate.
机译：解脂耶氏酵母脂肪酶Lip2（YlLip2）是酯合成，生物柴油生产和对映异构体拆分的重要生物催化剂。带有N端组氨酸标签的His6-YlLip2YlLip2成功在巴斯德毕赤酵母中表达。已经比较了三种不同的培养策略通过P生产His6-YlLip2的策略。结果显示，通过采用联合策略，可以大大提高“ His6-YlLip2”的活性和细胞存活率。“使用低盐培养基”（LSM）代替？碱性盐培养基（BSM）并将温度从28降低到25°C，His6-YlLip2的最大活性和容积生产率分别提高了55.3和79.8％。通过将山梨糖醇与甲醇一起共同进料，可以进一步提高His6-YlLip2的产量和细胞活力。在这种培养策略中，诱导114小时后，His6-YlLip2α的最大活性达到了15,600μU/ ml。甲醇诱导120h后，细胞死亡率降低了约11.2％（对照降低了约27.6％）。N末端组氨酸标签为纯化提供了方便。 His6-YlLip2的分子量约为38 kDa。当使用橄榄油作为底物时，纯的His6-YlLip2的比活度为4,830 U / mg。

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