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Home > Archive>Volume 37, Issue 12, 2018 >1278-1283. DOI:10.3969/j.issn.1673-1689.2018.12.008
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Improvement of TGase Thermal Stability through Site-Directed Mutagenesis Based on Analysis of Folding Free Energy
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    Abstract:

    Transglutaminases（EC 2.3.2.13，TGase） is an industrial enzyme widely used in several domains that include food processing and textile industry. The amino acid sites that may reduce folding free energy were predicted by using PoPMuSiC-2.1 to improve the thermal stability of TGase，and the corresponding mutants were then constructed. As indicated by PoPMuSiC-2.1 analysis，mutation at P132 revealed the highest level of decreases in folding free energy of TGase.Based on this prediction，the mutants with decreased folding free energy was thus contracted，including P132I，P132G，P132M，and P132Q. The analysis of enzymatic property showed that the half-life of P132I can reach 5.0 min at 50 ℃ which was 31% higher than that of wild-type TGase，and the other mutants also exhibited enhanced half-life. Moreover，P132I and P132G showed 24% and 12.4% enhanced specific activities in contrast to wild-type enzyme，respectively；while the specific activities of the rest mutants did not change significantly. And the structure analysis indicated that P132I mutant formed additional two hydrogen bonds as compared with wild-type enzyme. These results suggested that site-directed mutagenesis based on analysis of folding free energy is an effective method to improve the thermal stability of TGase，and newly formed hydrogen bond may be one of the reasons for enhanced thermal stability of P132I.

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TONG Liming, LIU Song, LI Jianghua, DU Guocheng, CHEN Jian. Improvement of TGase Thermal Stability through Site-Directed Mutagenesis Based on Analysis of Folding Free Energy[J]. Journal of Food Science and Biotechnology,2018,37(12):1278-1283.

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