Lü Ruihua, Shi Linna, Zhang Xirong, Feng Zhao
College of Medical Technology, Shaanxi University of Chinese Medicine, Xianyang 712046, Shaanxi, China.
Sheng Wu Gong Cheng Xue Bao. 2022 Jan 25;38(1):386-395. doi: 10.13345/j.cjb.210691.
Cutinase can degrade aliphatic and aromatic polyesters, as well as polyethylene terephthalate. Lack of commercially available cutinase calls for development of cost-effective production of efficient cutinase. In this study, eight cutinase genes were cloned from The most active gene was obtained by PCR combined with RT-PCR, expressed in BL21 and purified by Ni-NTA affinity chromatography to study its characteristics and pathogenicity. had a total length of 768 bp and 17 signal peptides at the N terminals. Phylogenetic analysis showed that its amino acid sequence had the highest homology with cutinase and was closely related to cutinase. was highly expressed during the process of infecting plants by . Moreover, the expression level of was higher than those of other cutinase genes in the process of sclerotia formation from mycelium. The heterologously expressed cutinase existed in the form of inclusion body. The renatured SsCut-52 was active at pH 4.0-10.0, and mostly active at pH 6.0, with a specific activity of 3.45 U/mg achieved. The optimum temperature of SsCut-52 was 20-30 ℃, and less than 60% of the activity could be retained at temperatures higher than 50 ℃. Plant leaf infection showed that SsCut-52 may promote the infection of Banlangen leaves by .
角质酶能够降解脂肪族和芳香族聚酯以及聚对苯二甲酸乙二酯。由于缺乏可商购的角质酶，因此需要开发具有成本效益的高效角质酶生产方法。在本研究中，从……克隆了8个角质酶基因。通过PCR结合RT-PCR获得了最具活性的基因，在BL21中表达并通过镍-亚氨基二乙酸亲和层析进行纯化，以研究其特性和致病性。……全长768 bp，N端有17个信号肽。系统发育分析表明，其氨基酸序列与……角质酶具有最高的同源性，并且与……角质酶密切相关。……在……感染植物的过程中高表达。此外，在菌丝体形成菌核的过程中，……的表达水平高于其他角质酶基因。异源表达的角质酶以包涵体的形式存在。复性后的SsCut-52在pH 4.0-10.0时具有活性，在pH 6.0时活性最高，比活性达到3.45 U/mg。SsCut-52的最适温度为20-30℃，在高于50℃的温度下活性保留不到60%。植物叶片感染实验表明，SsCut-52可能促进……对板蓝根叶片的感染。
