A novel mono- and diacylglycerol lipase highly expressed in Pichia pastoris and its application for food emulsifier preparation
A novel mono- and diacylglycerol lipase highly expressed in Pichia pastoris and its application for food emulsifier preparation
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DOI：
10.1016/j.procbio.2013.08.021
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发表时间：
2013-12-01
影响因子：
4.4
通讯作者：
Li, Ying
中科院分区：
文献类型：
--
作者：
Huang, Jinjin;Yang, Zhen;Li, Ying
从圆弧青霉（Penicillium cyclopium）中克隆出一种单酰甘油和二酰甘油脂肪酶（MDL），并在毕赤酵母（Pichia pastoris）菌株GS115中表达。该重组酶被命名为脂肪酶GH1。利用BSMG培养基在7.5升发酵罐中进行高细胞密度发酵，其中基础盐培养基中的磷酸盐被甘油磷酸钠（Na₂GP）替代。检测到的最大脂肪酶活性为每毫升18000单位，发酵上清液中的总蛋白质含量为每升3.94克。该液体酶在4℃碱性条件下活性稳定保持6个月，1年后活性为50%。脂肪酶GH1用于合成单酰甘油和二酰甘油（MAG和DAG），它们是工业应用中常用的乳化剂。在甘油/油酸摩尔比为11∶1、含水量为1.5%（重量比）、酶用量为每克80单位以及反应温度为35℃的条件下，反应24小时后转化率达到84%。当使用油酸作为酰基供体时，脂肪酶GH1在合成MAG和DAG方面比脂肪酶G50（一种类似的、市售的来源于卡门培尔青霉（Penicillium camemberti）的脂肪酶）更高效。脂肪酶GH1在食品乳化剂制备方面具有潜力。（C）2013爱思唯尔有限公司。保留所有权利。
A mono- and diacylglycerol lipase (MDL) was cloned from Penicillium cyclopium and expressed in Pichia pastoris strain GS115. The recombinant enzyme was named Lipase GH1. High cell density fermentation was performed by culture in a 7.5-L fermenter using BSMG medium, in which the phosphate in basal salt medium was replaced by sodium glycerophosphate (Na(2)GP). The maximal lipase activity detected was 18,000U per mL, and total protein content in the fermentation supernatant was 3.94g per L The activity of the liquid enzyme remained stable under alkaline conditions at 4 degrees C for 6 months and was 50% after one year. Lipase GH1 was used for the synthesis of mono- and diacylglycerols (MAGs and DAGs), which are commonly used emulsifiers for industrial applications. A conversion rate of 84% after 24 h of reaction was obtained using glycerol/oleic acid molar ratio 11:1, water content 1.5 wt%, enzyme dosage 80U per g, and reaction temperature 35 C. Lipase GH1 was more efficient for the synthesis of MAGs and DAGs than was Lipase G50 (a similar, commercially available lipase derived from Penicillium camemberti) when oleic acid was used as an acyl donor. Lipase GH1 has potential for food emulsifier preparation. (C) 2013 Elsevier Ltd. All rights reserved.