Department of Chemical Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan.
Department of Chemical Engineering, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan.
J Biotechnol. 2019 Apr 20;296:1-6. doi: 10.1016/j.jbiotec.2019.02.009. Epub 2019 Mar 7.
In this study, a commercial lipase derived from Candida cylindracea was chemically modified with dextran by conjugating ε-amine in the lysine residue with the carbonyl residue in oxidized dextran using the borane-pyridine complex as a reducing agent to increase the hydrophilicity of the microenvironment around the lipase in the presence of organic solvents. The degree of modification (53.2%), amount of dextran (0.66 g/g-lipase), specific activity (similar to that of the unmodified lipase), and stability in the presence of ethanol and 2-propanol (the half-lives were 2.24 and 1.86 times longer than those of the unmodified lipase) were higher for the lipase modified at pH 8.0 than for the lipases modified at other pH levels. Following modification with dextran at pH 8.0, the stability of the modified lipase was higher than that of the unmodified lipase in the presence of 25% (v/v) DMSO, ethanol, 2-propanol, toluene, n-hexane, and isooctane (the half-lives were 1.45, 2.24, 1.86, 1.76, 2.67 and 2.95 times longer than those of the unmodified lipase). Therefore, chemical modification with polysaccharides such as dextran using the borane-pyridine complex as a reducing agent could be a promising approach for improving the organic solvent stability of enzymes.
在这项研究中,通过使用硼烷-吡啶复合物作为还原剂,将 Candida cylindracea 来源的商业脂肪酶与氧化葡聚糖的羰基残基在赖氨酸残基上的 ε-胺进行缀合,从而化学修饰了葡聚糖,以增加有机溶剂存在下脂肪酶周围微环境的亲水性。在 pH 值为 8.0 时进行修饰的脂肪酶的修饰程度(53.2%)、葡聚糖量(0.66 g/g-脂肪酶)、比活性(与未修饰的脂肪酶相似)和在乙醇和 2-丙醇中的稳定性(半衰期比未修饰的脂肪酶长 2.24 和 1.86 倍)均高于在其他 pH 值下进行修饰的脂肪酶。在 pH 值为 8.0 时用葡聚糖进行修饰后,在 25%(v/v)DMSO、乙醇、2-丙醇、甲苯、正己烷和异辛烷存在下,修饰脂肪酶的稳定性均高于未修饰的脂肪酶(半衰期比未修饰的脂肪酶长 1.45、2.24、1.86、1.76、2.67 和 2.95 倍)。因此,使用硼烷-吡啶复合物作为还原剂对多糖(如葡聚糖)进行化学修饰可能是提高酶的有机溶剂稳定性的一种有前途的方法。