末端融合双亲短肽提高谷氨酰胺转胺酶热稳定性
首发时间:2016-05-23
摘要:谷氨酰胺转胺酶(Transglutaminase, EC 2.3.2.13, TGase)广泛应用于食品、纺织等领域。为提高TGase的热稳定性,将来源于Saccharomy cescerevisiae Zuotion蛋白的双亲短肽SAP1融合至TGase前导肽C端、成熟酶N端及C端,分别得到TGase融合酶QC-SAP1、N-SAP1和C-SAP1;与野生酶相比,N-SAP1在50℃下的半衰期提高78.9%,其它三个突变体则提高5-24%;各突变体比酶活下降4-30.3%。结构分析显示,与野生酶相比,N-SAP1的α-螺旋结构增加,蛋白颗粒较野生酶增大。上述结果表明,末端融合双亲短肽SAP1能有效的提高TGase的热稳定性,酶蛋白的二级结构的稳定化及寡聚化可能是融合酶热稳定性提高的重要原因。
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Enhancement of the thermal stability of transglutaminase through fusing amphipathic peptides at the terminus
Abstract:Transglutaminases (EC 2.3.2.13, TGase) is an industrial enzyme widely used in food processing, textile industry, and etc. To improve the thermal stability of TGase, the amphipathic peptides from Saccharomy cescerevisiae Zuotion (SAP1) was fused at C-terminus of its pro-peptide, N-terminus and C-terminus of the TGase, yielding the fusions QC-SAP1, N-SAP1, and C-SAP1, respectively. In contrast to the wild-type TGase, the half-life of N-SAP1 was 78.9% higher and the other mutants exhibited 5-24% enhanced half-life. As indicated by circular dichroism spectrum and transmission electron microscope, N-SAP1 showed an increase in a-helix structure and a bigger particle in comparison with the wild-type enzyme. The results suggested that fusing with the amphipathic peptides at N-termius can effectively enhance the thermal stability of TGase, and the stabilization of the secondary structure and oligermization of the enzyme protein may be one of the most important reason for the enhancement.
Keywords: transglutaminases thermal stability amphipathic peptide terminal fusion
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