Enzymatic Properties of Recombinant Ligase Butelase-1 and Its Application in Cyclizing Food-Derived Angiotensin I-Converting Enzyme Inhibitory Peptides

Butelase-1 is an efficient Ligase from Clitoria ternatea with wide applications in the food and biopharmaceutical fields. This research aimed to achieve high-efficiency expression of butelase-1 and explore its application in food-derived angiotensin I-converting Enzyme (ACE) inhibitory Peptides. The recombinant butelase-1 zymogen was prepared at a yield of 100 mg/L in Escherichia coli and successfully activated at pH 4.5, resulting in a 6973.8 U/L yield of activated butelase-1 with a specific activity of 348.69 U/mg and a catalytic efficiency of 9956 M^-1 s^-1. Activated butelase-1 exhibited considerable resistance to Tween-20, Triton X-100, and methanol. The "traceless" cyclization of ACE inhibitory Peptides was realized using activated butelase-1, which resulted in higher stability and ACE inhibitory activity than those of the linear Peptides. Our work proposed an efficient method for the preparation of butelase-1 and provided a promising model for its application in food fields.

ACE inhibitory peptides; butelase-1; enzymatic cyclization; enzymatic properties; heterologous expression.