Development of a Pichia pastoris whole-cell biocatalyst with overexpression of mutant lipase I PCLG47I from Penicillium cyclopium for biodiesel production,RSC Advances

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Development of a Pichia pastoris whole-cell biocatalyst with overexpression of mutant lipase I PCLG47I from Penicillium cyclopium for biodiesel production
RSC Advances ( IF 3.9 ) Pub Date : 2018-07-20 00:00:00 , DOI: 10.1039/c8ra04462g
Yihan Liu _{1,

2,

3,

4} , Lin Huang _{1,

2,

4} , Dong Zheng _{1,

4} , Yu Fu _{1,

4} , Mengying Shan _{2,

4} , Zehua Xu _{2,

4} , Jieying Ma _{2,

4} , Fuping Lu _{1,

2,

3,

4}

Affiliation

Penicillium cyclopium lipase I (PCL) is a thermolabile triacylglycerol lipase with very low activity against monoacylglycerols, and there have been no reports on the transesterification of oil to produce biodiesel. A mutant PCL^G47I with an improved thermostability was previously obtained through replacing Gly47 with Ile in PCL. In this study, a novel Pichia pastoris whole-cell biocatalyst (WCB) with overexpression of PCL^G47I was constructed and characterized for biodiesel production from soybean oil. The optimum conditions for biodiesel preparation were 1 g soybean oil, 1 : 2 initial oil/methanol molar ratio with 3 times methanol addition of 1 : 0.75 oil/methanol molar ratio at 4 h intervals, 7% water content, 400 U lipase, temperature of 25 °C, and reaction time of 20 h. Under the optimum conditions, the FAME yield reached 60.7% and remained 47.3% after 4 batch cycles, and no glycerol was generated as a byproduct. These findings indicated that this WCB is a promising biocatalyst for biodiesel production in a relatively cost-effective manner. Additionally, the resulting enzymatic process may provide a potential method for biodiesel production at an industrial scale.

中文翻译：

从青霉中过表达突变脂肪酶 I PCLG47I 的毕赤酵母全细胞生物催化剂的开发用于生物柴油生产

环青霉脂肪酶 I (PCL) 是一种不耐热的三酰基甘油脂肪酶，对单酰基甘油的活性非常低，并且没有关于油的酯交换生产生物柴油的报道。先前通过在 PCL 中用 Ile 代替 Gly47 获得了具有改善的热稳定性的突变 PCL ^{G47I 。}在本研究中，一种新型毕赤酵母全细胞生物催化剂 (WCB) 过表达 PCL ^G47I构建和表征用于从大豆油生产生物柴油。制备生物柴油的最佳条件为 1 g 大豆油、1:2 初始油/甲醇摩尔比、3 次甲醇添加 1:0.75 油/甲醇摩尔比 (4 h 间隔)、7% 含水量、400 U 脂肪酶、温度25 °C，反应时间 20 h。在最佳条件下，FAME 收率达到 60.7%，经过 4 个批次循环后仍保持在 47.3%，并且没有生成甘油作为副产物。这些发现表明，这种 WCB 是一种很有前途的生物催化剂，可用于以相对具有成本效益的方式生产生物柴油。此外，由此产生的酶促过程可以为工业规模的生物柴油生产提供潜在的方法。

更新日期：2018-07-20

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