The N-terminal hydrophobicity modulates a distal structural domain conformation of zearalenone lacton hydrolase and its application in protein engineering

Wang, Hui; Lu, Zhenghui; Lin, Xiangui; Wang, Meixing; Jiang, Tianzhi; Zhao, Guoqiang; Xiang, La; Xv, Jiazhan; Jiang, Sijing; Zhang, Guimin

doi:10.1016/j.enzmictec.2023.110195

Cited by 4 publications

(5 citation statements)

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“…The authors of ZHD11C hypothesized that, based on the results of molecular dynamics simulations, introducing a single hydrophobic residue at the N-terminus is likely to affect the distal conformation of the entire protein. 23 Nevertheless, they did not explain the mechanism of this putative phenomenon. Dumon concluded that the residues introduced at the N-terminus slow the increase in entropy during protein unfolding.…”

Section: ■ Results and Discussionmentioning

confidence: 98%

“…Wang et al reported the hydrophobic residue at the N-terminus to be the key factor that gives ZHD11C good thermal stability, and the introduction or deletion of a single hydrophobic residue at the N-terminus can have a significant effect on the thermal stability of the enzyme. 23 In this study, two mutations were designed for the N-terminal region of ZENY. The first 11 amino acids of the N-terminus were replaced with the first 13 amino acids of the N-terminal region of ZHD11C (Figure 7).…”

Section: ■ Results and Discussionmentioning

confidence: 99%

“…Truncation of these N-terminal sequences results in misfolding and reduced or lost activity. Wang et al reported the hydrophobic residue at the N-terminus to be the key factor that gives ZHD11C good thermal stability, and the introduction or deletion of a single hydrophobic residue at the N-terminus can have a significant effect on the thermal stability of the enzyme . In this study, two mutations were designed for the N-terminal region of ZENY.…”

Section: Resultsmentioning

confidence: 99%

“…However, this is not a good explanation for the increased thermal stability. The authors of ZHD11C hypothesized that, based on the results of molecular dynamics simulations, introducing a single hydrophobic residue at the N-terminus is likely to affect the distal conformation of the entire protein . Nevertheless, they did not explain the mechanism of this putative phenomenon.…”

Section: Resultsmentioning

confidence: 99%

“…ZEN lactone hydrolases are generally associated with low adaptability to temperature changes, and their activity decreases significantly or is eliminated when the temperature exceeds 50 °C. RmZHD and ZHD11C, which have better heat resistance, exhibit optimal degradation activity at 45 °C. The optimal reaction temperature of ZENY was 37 °C; the activity at 45 °C was approximately 70%, and only 30% activity remained at 50 °C (Figure B).…”

Section: Resultsmentioning

confidence: 99%

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Characterization, Structural Analysis, and Thermal Stability Mutation of a New Zearalenone-Degrading Enzyme Mined from Bacillus subtilis

Shi,

Mwabulili,

Xie

et al. 2024

J. Agric. Food Chem.

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Zearalenone (ZEN) is a widespread mycotoxin that causes serious damage to animal husbandry and poses a threat to human health. A screen of ZEN-degrading soil bacteria yielded Bacillus subtilis YT-4, which yielded 80% ZEN degradation after 6 h and 95% after 36 h. The gene sequence encoding the degradative enzyme ZENY was mined from the genome of YT-4 and expressed in yeast. ZENY is an α/β-hydrolase with an optimal enzyme activity at 37 °C and pH 8. By breaking the lactone ring of ZEN, it produces ZENY-C 18 H 24 O 5 with a molecular weight of 320.16 g/mol. Sequence comparison and molecular docking analyses identified the catalytic ZENY triad 99S-245H-123E and the primary ZEN-binding mode within the hydrophobic pocket of the enzyme. To improve the thermal stability of the enzyme for industrial applications, we introduced a mutation at the N-terminus, specifically replacing the fifth residue N with V, and achieved a 25% improvement in stability at 45 °C. These findings aim to achieve ZEN biodegradation and provide insight into the structure and function of ZEN hydrolases.

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Section: ■ Results and Discussionmentioning

confidence: 98%

Section: ■ Results and Discussionmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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