Intercollegiate Faculty of Biotechnology, University of Gdansk and Medical University of Gdansk, Abrahama 58, 80-307, Gdańsk, Poland.
Department of Plant Physiology and Biotechnology, Faculty of Biology, University of Gdańsk, Wita Stwosza 59, 80-308, Gdańsk, Poland.
BMC Plant Biol. 2021 Jan 19;21(1):50. doi: 10.1186/s12870-020-02823-4.
Simmondsia chinensis (jojoba) is the only plant known to store wax esters instead of triacylglycerols in its seeds. Wax esters are composed of very-long-chain monounsaturated fatty acids and fatty alcohols and constitute up to 60% of the jojoba seed weight. During jojoba germination, the first step of wax ester mobilization is catalyzed by lipases. To date, none of the jojoba lipase-encoding genes have been cloned and characterized. In this study, we monitored mobilization of storage reserves during germination of jojoba seeds and performed detailed characterization of the jojoba lipases using microsomal fractions isolated from germinating seeds.
During 26 days of germination, we observed a 60-70% decrease in wax ester content in the seeds, which was accompanied by the reduction of oleosin amounts and increase in glucose content. The activity of jojoba lipases in the seed microsomal fractions increased in the first 50 days of germination. The enzymes showed higher activity towards triacylglycerols than towards wax esters. The maximum lipase activity was observed at 60 °C and pH around 7 for triacylglycerols and 6.5-8 for wax esters. The enzyme efficiently hydrolyzed various wax esters containing saturated and unsaturated acyl and alcohol moieties. We also demonstrated that jojoba lipases possess wax ester-synthesizing activity when free fatty alcohols and different acyl donors, including triacylglycerols and free fatty acids, are used as substrates. For esterification reactions, the enzyme utilized both saturated and unsaturated fatty alcohols, with the preference towards long chain and very long chain compounds.
In in vitro assays, jojoba lipases catalyzed hydrolysis of triacylglycerols and different wax esters in a broad range of temperatures. In addition, the enzymes had the ability to synthesize wax esters in the backward reaction. Our data suggest that jojoba lipases may be more similar to other plant lipases than previously assumed.
西蒙得木(荷荷芭)是唯一已知在其种子中储存蜡酯而不是三酰基甘油的植物。蜡酯由非常长链的单不饱和脂肪酸和脂肪醇组成,占荷荷芭种子重量的 60% 左右。在荷荷芭种子萌发过程中,蜡酯动员的第一步是由脂肪酶催化的。迄今为止,尚未克隆和表征任何荷荷芭脂肪酶编码基因。在这项研究中,我们监测了荷荷芭种子萌发过程中储存物质的动员情况,并使用从萌发种子中分离的微粒体部分对荷荷芭脂肪酶进行了详细的表征。
在 26 天的萌发过程中,我们观察到种子中蜡酯含量下降了 60-70%,同时油体数量减少,葡萄糖含量增加。种子微粒体部分的荷荷芭脂肪酶活性在萌发的前 50 天增加。与蜡酯相比,这些酶对三酰基甘油的活性更高。最大脂肪酶活性在 60°C 和 pH 约 7 时观察到,对三酰基甘油而言,对蜡酯而言则为 6.5-8。该酶有效地水解了各种含有饱和和不饱和酰基和醇部分的蜡酯。我们还证明,当使用游离脂肪酸和不同的酰基供体,包括三酰基甘油和游离脂肪酸作为底物时,荷荷芭脂肪酶具有蜡酯合成活性。对于酯化反应,该酶利用饱和和不饱和脂肪醇,优先使用长链和超长链化合物。
在体外测定中,荷荷芭脂肪酶在较宽的温度范围内催化三酰基甘油和不同蜡酯的水解。此外,这些酶在反向反应中具有合成蜡酯的能力。我们的数据表明,荷荷芭脂肪酶可能比以前认为的更类似于其他植物脂肪酶。