Yan Wei, Li Furong, Wang Li, Zhu Yaxin, Dong Zhiyang, Bai Linhan
Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, China.
State Key Laboratory of Microbial Resources, Institute of Microbiology Chinese Academy of Science, Beijing, 100101, China.
Biotechnol Rep (Amst). 2016 Dec 23;14:27-33. doi: 10.1016/j.btre.2016.12.007. eCollection 2017 Mar.
A new gene encoding a lipase (designated as ) was identified from a metagenomic bacterial artificial chromosome(BAC) library prepared from a concentrated water sample collected from a hot spring field in Niujie, Eryuan of Yunnan province in China. The open reading frame of this gene encoded 622 amino acid residues. It was cloned, fused with the oleosin gene and over expressed in to prepare immobilized lipase artificial oil body AOB-sole-lip-1. The monomeric Sole-lip-1 fusion protein presented a molecular mass of 102.4 kDa. Enzyme assays using olive oil and methanol as the substrates in petroleum ether confirmed its transesterification activity. Hexadecanoic acid methyl ester, 8,11-Octadecadienoic acid methyl ester, 8-Octadecenoic acid methyl ester, and Octadecanoic acid methyl ester were detected. It showed favorable transesterification activity with optimal temperature 45 °C. Besides, the maximal biodiesel yield was obtained when the petroleum ether system as the organic solvent and the substrate methanol in 350 mmol/L (at a molar ratio of methanol of 10.5:1) and the water content was 1%. In light of these advantages, this lipase presents a promising resource for biodiesel production.
从中国云南省洱源县牛街温泉田采集的浓缩水样制备的宏基因组细菌人工染色体（BAC）文库中，鉴定出一个编码脂肪酶的新基因（命名为 ）。该基因的开放阅读框编码622个氨基酸残基。将其克隆，与油质蛋白基因融合，并在 中过量表达，以制备固定化脂肪酶人工油体AOB-sole-lip-1。单体Sole-lip-1融合蛋白的分子量为102.4 kDa。在石油醚中以橄榄油和甲醇为底物进行酶活性测定，证实了其酯交换活性。检测到十六烷酸甲酯、8,11-十八碳二烯酸甲酯、8-十八碳烯酸甲酯和十八烷酸甲酯。它在45℃的最佳温度下表现出良好的酯交换活性。此外，当以石油醚体系作为有机溶剂，底物甲醇浓度为350 mmol/L（甲醇摩尔比为10.5:1）且水含量为1%时，生物柴油产量最高。鉴于这些优势, 这种脂肪酶是生物柴油生产的一个有前景的资源。
